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1. FL1 FL2 FL3 FL4 FL5 fluorescence detector to correct for a THET the overlap of one dye s emission into 147 15 2 13 99 another dye s detector 49 35 4 3 0 48 Compensation Matrix Ye 24 Clear All Button op a 0 0 3s Select the column which contains the Fluorochrome creating spectral overlap in the PMT eas row Clear All wo See also Cytometer Control Cytometer Control Acquisition Setup Tab Cytometer Control Detector Tab Compensation Matrix Click in the box and type a value or use the Slide Bar to set the desired value Use the Up and Down arrows to make any fine adjustments to the value Clear All Button Selecting the Clear All button resets ALL the values within the Compensation Matrix to zero PN 177579BD USING MXP SOFTWARE CYTOMETER MENU Status Page The Status Page displays the x Cytometer settings currently in use and allows you to print these if i required Fs _ss rit FL2 rts FLa Fis aux 412 499 0 Print i Selecting the Print button displays the standard Windows Print dialog box iscriminator L5 aser Power mW Cytometer Log Use this menu item to view the log of errors generated by the system It also allows you to clear this log as needed Get Cytosettings from Protocol The Get Cytosettings From Protocol feature allows Cytometer Instrument Settings to be obtained from
2. Cytometer Serial No Greann Laboratory COULTER Cytomics FC 500 MPL 7579024B PN 177579BD A 7 LOG SHEETS A 8 PN 177579BD Application Flow Set Fluorospheres Flow Set 675 Fluorospheres Flow Set 770 Fluorospheres Mode Target Ranges FS SS LOG SS LOG SHEETS A ESTABLISHING HV TOTAL GAIN RANGES Lot Number Lot Number Lot Number LOG FL1 Expiration Date Expiration Date Expiration Date LOG FL3 LOG FL2 LOG FL4 LOG FL5 FS SS or LOG SS LOG FL1 LOG FL2 LOG FL3 LOG FL4 LOG FL5 Mode HV a 5 Red laser CO CO NI gt On amp oo po Average HV Gain Average 2SD or 1 Average 2SD or 1 Cytometer Serial No Laboratory Cytomics FC 500 MPL PN 177579BD COULTER 7579025B A 9 LOG SHEETS A 10 PN 177579BD DAILY LOG FOR INSTRUMENT VERIFICATION OF ALIGNMENT AND FLUIDICS Flow Check Fluorospheres Lot Number Flow Check 675 Fluorospheres Lot Number Flow Check 770 Fluorospheres Lot Number Expiration Date Expiration Date Expiration Date LOG SHEETS A FS FL1 FL2 FL3 Target Range FL4 Red laser 488 a Run Mode HPCV Mode HPCV Mode HPCV Mode HPCV Mode HPCV Le He
3. USB box Solid state red laser UMBILICAL m Power Supply POWER MODULE CONTROL i F module ac power line cords 7579002B PN 177579BD 2 3 INSTALLATION SYSTEM CONNECTIONS Figure 2 2 Power and Signal Cable Connections with Universal Power Supply Cytometer back cover Cytometer Workstation z z USB box T EE ta Fiber optic trans to rec N A Fiber optic rec to xmit Solid state red laser Logic Ae A f Tr Power nalog module a control a a ll ac power Power line cords Supply Laser umbilica module 7579046B PN 177579BD 2 4 INSTALLATION 2 SYSTEM CONNECTIONS Waste and Pneumatic Tubing Figure 2 3 or Figure 2 4 shows the interunit connections for waste and pneumatic tubing depending upon your Power Supply configuration Note Ensure the waste tubing does not exceed the height of the MPL sample probe Note Place the 10 L external sheath fluid container in a location that is lower than the internal sheath fluid container This prevents siphoning of the sheath fluid Figure 2 3 Waste and Pneumatic Tubing Connections with Voltage Specific P
4. When the Login dialog box appears enter your User ID and your password C Note If you reach the limit of password attempts allowed you are not able to log into the MXP software You must ask the System Administrator to unlock you and or reset your password before you can log into MXP software x Username Gladstone bd Cancel PN 177579BD USING MXP SOFTWARE WINDOW MENU 8 If Yes Click Here appears in the Signed field of a version you can J Yes Click Here to view the signature information for that version after you review the signatures 9 Repeat the procedure for each version to be signed Changed Signed Comments 13 Oct 2004 12 13 10 added stats displayed 13 Oct 2004 12 12 55 Yeas Click Hara printed PDFs 13 Oct 2004 12 12 47 Yes Click Here exported data Filename 07_5c 21 as verify 8 19 3 45 4 Imd Version 3 Number of Signatures 3 Signature Completed a Signed by Gladstone Signed on 13 Oct 2004 12 20 17 Signature Approved Signed by Gladstone Signed on 13 Oct 2004 12 22 24 Signature Certified Signed by Gladstone Signed on 13 Oct 2004122251 Z 10 Close when you are done B 24 WINDOW MENU Cascade Use Cascade to display the window so that the plots overlay WD Window gt Cascade Tile Arranges each plot so every window display is visible The pattern depends on the number of open Windows W Window gt gt
5. 11 Select Analysis Select Results 12 You must select FCS Keywords SampleID1 through SampleID4 Statistic Type p Report Options Curent Filename Only select the statistics required for Bos hens data reduction Note No matter how many statistics ieee you select at this screen only these nine statistics are used for data analysis TESETTINGSHLENT BEAD HPPO and graphing by either of the Analysis Mae se j Dea je Cell uL Number Gated X Mean Y Mean X Mode Y Mode X Median Y Median Note Proceed to step 19 unless you are setting up to send files during data acquisition 13 If calibrators are included in your assay setup then for each calibrator a Assign Calibrator as the Well Type b In Sample Id enter the actual concentration pL of the standard c Enter the Dilution factor as appropriate Well Information Well Type Sample Id Dilution Replicates Calibrator v 50 10 Ho B 19 B USING MXP SOFTWARE FILE MENU B 20 14 After you define and save the plate definition at the Plate Wizard J Lr to start acquisition 15 Do not edit Sample ID 2 3 or 4 in the Worklist when using either of the Analysis Add Ins 16 MS Excel automatically opens during data acquisition of the sample plate 17 During acquisition do not modify the protocol including region or gate adjustments or the data might be divided into multiple sheets 18 If you have the 21 CFR P
6. Cancel Help See also Toolbars Customize Toolbars Categories Displays the Toolbars that are currently available Buttons Shows the buttons that may be selected for any Toolbar Description Displays a description of the Icon button selected Zoom The following option allows you to zoom in on a FlowPAGE to 100 200 or 400 Select the desired view Selecting 100 returns to the default size Refresh Selecting View Refresh or pressing F5 causes MXP software to refresh the screen all plots and FlowPAGEs This option is useful when Windows fails to refresh the screen correctly leaving some graphical errors visible PN 177579BD B 73 B USING MXP SOFTWARE INSERT MENU B 17 B 74 INSERT MENU Blank FlowPAGE To open a new blank FlowPAGE D al Note e The page displayed in the FlowPAGE is the same aspect ratio and paper orientation as used in the default Printer e The page scaling can be changed at any time after a FlowPAGE is created e Several FlowPAGEs can be created To close or delete a FlowPAGE double click on the Control menu box in the upper left corner of the FlowPAGE window to be deleted All FlowPAGEs contain e Institution name e Acquisition or playback protocol e Acquisition date and time e User ID e Settings file name date and time e Listmode file name e Total acquisition time and stop condition e Sample ID 1 e Protocol status e Analysis date e Instrument
7. The combinations of dividers are combined to produce 2P regions 2 regions Where p is the number of prism dividers set PN 177579BD B 113 USING MXP SOFTWARE PLOTS MENU This table shows the number of phenotypes created for up to six Prism dividers set Number of Prism Number of Dividers set phenotypes created This Plot shows a 3 color Prism Plot 3 dividers set 8 phenotypes created The percentages printed on the prism plot from left to right match one to one with the phenotype nomenclature indicated on the axes Example Triple Positive phenotype is 0 7 Triple Negative phenotype is 25 1 The phenotype percentages are staggered to prevent excessive obscuration of the numerical values when a high density of prism information is presented Editing Prism Dividers 1 Mouse over the divider to get the cursor 2 D and drag the divider to the new position B 114 PN 177579BD USING MXP SOFTWARE B PLOTS MENU Removing Prism Dividers 1 D the Prism divider you want to delete 2 Press Delete MXP Software Prism Compatibility Note Reanalysis of listmode files overrides the analysis protocol s Prism dividers You have to adjust the dividers when this occurs Third party software may not display Prism data correctly since a special Prism data display algorithm is involved to display Prism in MXP software PN 177579BD B 115 USING MXP SOFTWARE PLOTS MENU B
8. DaCS Users For Use Outside MXP Software The System Administrator must set up Users in the DaCS software so the Users can have access to additional functionality such as electronic signatures during analysis of Excel data files when MXP software is not open PN 177579BD C 6 PN 177579BD GLOSSARY 21 CFR Part 11 The FDAs Electronic Records and Electronic Signatures Rule that outlines the requirements for submitting documentation in electronic form and the criteria for approved electronic signatures is also known as 21 CFR Part 11 It defines the standards by which an organization can use electronic records to meet its record keeping requirements The MXP software has a 21 CFR Part 11 option that enables your laboratory to comply with this regulation Accuracy The ability of an instrument to agree with a predetermined reference value at any point within the operating range Contrast with precision APC Abbreviation for allophycocyanin dye ASCII Abbreviation for American Standard Code for Information Interchange An ASCII file is a type of text file Assay values Values for a control established by extensive repeat testing of that control AutoGate A gate that automatically sizes to the population contained within the boundaries of the region There are two types Elliptical and Contour AutoSetup Wizard A software feature that guides you through the processing of quality control materials to automatically esta
9. The System Administrator sets the time interval when the User Password expires and must be changed The System Administrator sets the time period of user inactivity that causes the User to be automatically logged out of the MXP software The System Administrator sets the number of unsuccessful password attempts that locks a user out of the system Only the System Administrator can unlock the user and if needed reset the password The System Administrator can view and print Audit Trails that include login activities password changes and failed password attempts for each current and deleted user There is to be no unauthorized use of another user s ID or password Automatic and Manual Lock Out MXP software provides automatic and manual lock out procedures to control access to the system Only a logged in user or the System Administrator should have access to the MXP software Automatic Lock Out The System Administrator defines the time limit of user inactivity no keyboard or mouse action that causes the User to be automatically logged out of the MXP software Once logged out the user must log in at the Startup Wizard screen to access the MXP software If automated activities are in progress when automatic log out would normally occur 1 these activities continue without interruption 2 the MXP software is not exited but all toolbars and menus are disabled except the File gt Log In menu item and the Cytometer Abort Acquisition
10. DATE 29 Jul 96 DFC1TO2 17 5 DFC2TO1 0 6 DFC2TO3 38 6 DFC3TO2 13 2 DFC3T04 40 7 DFC4T02 2 8 DFC4T03 0 9 FIL JO003946 LMD INST Coulter Technology Center xl display the FCS Keyword as well as the associated keyword value Multiple Keywords can be selected from the scroll box Select Clear All Selected to deselect the items Note Information is displayed in the FCS dialog box only when a valid FCS listmode file is open or has just been acquired See also Legend Data Source Legend Legend Options Tab Format Plot PN 177579BD USING MXP SOFTWARE B PLOTS MENU Acquisition Stop and Save Use Acquisition Stop and Save to terminate the acquisition of a sample at a preset stop value If there is more than one stop value set then the first stop counter reached terminates the current acquisition Note The values set in the Cytometer Control Acquisition Limits duration and Max events overrides the stop values set here in the event of an acquisition limit being reached before the Acquisition Stop value has been achieved Single Parameter Histograms can be selected for automatic saving in FCS Format Note An acquisition from a tissue culture sample is to be stopped on 5 000 cells to allow the collection and storage of all event data In a tissue culture sample with two cell types where cell type A accounts for about 30 of the white cell count we have no problem with the above
11. Note To avoid unnecessary DaCS errors when launching the MXP software and to allow access to all file folders necessary for 21 CFR Part 11 read write functionality set up Windows User accounts with full Administrator rights on a FC 500 MPL flow cytometer that has a Workstation with MXP 21 CFR Part 11 software installed Third Party Software Applications PN 177579BD If you use any third party software application such as Adobe Acrobat you need to have procedures that ensure the security of these electronic records and provide an audit trail Security software applications may be purchased to provide security when using third party software applications C 3 fe 21 CFR PART 11 OPTION COMPLIANCE INFORMATION ARCHIVING AND RESTORING ARCHIVED DATA C 3 C 4 C 5 C 6 C 4 ARCHIVING AND RESTORING ARCHIVED DATA The System Administrator has the ability to archive data Archived data can also be restored for additional analysis by the System Administrator Whenever a user tries to open an archived data file that has not been restored the user is notified by the software of the archive directory path and directed to request the System Administrator to restore the archived file needed A restored file is restored into the MXP software as a new record for 21 CFR Part 11 purposes Its version number becomes number one once it is opened for use AUDIT TRAILS Only the System Administrator can view and print the Audit Trails
12. 15 94 in 17 25 in 48 4 lb Power Supply 58 4 cm 45 7 cm 71 1 cm 59 0 kg Universal 23 in 18 in 28 in 130 lb Power Supply 50 8 cm 45 7 cm 66 0 cm 54 4 kg Voltage 20 in 18 in 26 in 120 lb specific This is the width with a single argon laser or with the optional red solid state laser The width of the FC 500 MPL with the optional HeNe laser is 119 4 cm 47 in Installation Category Category II per IEC 1010 1 standard Noise lt 65 db Cytometer Flow Cell Sensing area BioSense 150 pm x 450 rectangular channel with an integral lens mounted with a vertical upward flow path PN 177579BD 3 1 SPECIFICATIONS INSTRUMENT SPECIFICATIONS 3 2 Flow Rate Continuous pressure is applied to the sample pickup tubing The amount of pressure depends on the flow rate you specify e Low 10 pL min e Medium 30 pL min e High 60 pL min Lasers Air cooled software controlled 20 mW Argon ion laser operating at 488 nm The optional second laser is a solid state software controlled 25 mW diode laser operating nominally at 635 nm The second laser has a shutter that allows you to block the laser output to the flow cell Argon Laser Power Laser power is monitored 5 times per second within the system software and the reading is displayed on the STATUS page of the MXP cytometer software If the laser power deviates more than 1 a Laser Power Error 4206 is displayed on the Status Bar and t
13. 6 W to accept changes does not apply any of the changes made Note To select all the files press ctri A Status Icon 7 Pause The Auto Pause function pauses the run of tests and then carries on with the rest of the panel automatically after a 15 second wait The countdown shows on the AutoMATOR Status Bar When the tests are in the Auto Pause setting you can hold down and click the Pause u button overrides the Auto Pause placing the AutoMATOR into Continuous Pause To restart click the Run gt button and the test resumes from the point at which it was paused any Continuous Pause Pauses for modifications and remains on pause until you decide that processing will continue This function allows the test to stop without reverting to the beginning of the panel again To resume the test click the Run button 4 Continue Plays back the indicated file without pausing B 86 PN 177579BD PN 177579BD USING MXP SOFTWARE TOOLS MENU These icons allow you to insert a pause or a stop into the panel or let the panel run straight through from start to finish Clicking repeatedly on the icon until it displays the desired status modifies the status in the highlighted file AMV Well 17 Specimen R_1 Undefined vandy 270 LMD 4A19_Well 19 Specimen R_1 Undefined vandy 271 LMD 421_Well 21 Specimen R_1 Undefined vandy 272 LMD 3 B 23_Well 23 Specimen R_1 Undefined vandy 273 LMD_ In this example the four files would
14. E EEEE N gt gt 18 P e F B 10 Create a new Worklist clear Worklist Save a Panel Accesses the appropriate Save Panels dialog box depending upon whether you have standard MXP software or the 21 CFR part 11 option installed Add a Panel to the Worklist Accesses the appropriate Open Playback Panels dialog box depending upon whether you have standard MXP software or the 21 CFR part 11 option installed Add a Well to the Worklist Delete a Well from the Worklist Play Pause Stop Sort the Worklist Minimize the Listmode Playback window Load Playback Worklist Save Playback Worklist Load Listmode Queue Accesses the appropriate Open Playback Panels Files dialog box depending upon whether you have standard MXP software or the 21 CFR part 11 option installed Save Listmode Queue Accesses the appropriate Save Playback Panels Files dialog box depending upon whether you have standard MXP software or the 21 CFR part 11 option installed Replay Plate Worklist PN 177579BD USING MXP SOFTWARE B FILE MENU B 14 FILE MENU New Protocol Use this option to create a New Protocol 1 w File gt New gt gt New Protocol The Save Existing Protocol dialog box is displayed 2 Ifyou O Yes the Save Protocol File As dialog box is displayed O the folder in which you wish to Save the Protocol Enter a valid File Name in File Name box and UP 3 Ifthe File Name already exists a warning box is displayed e I
15. HPCV Tech Date CO CO N on Boo po Cytometer Serial No Laboratory Cytomics FC 500 MPL PN 177579BD BECKMAN COULTER 7579026B A 11 LOG SHEETS A 12 PN 177579BD LOG SHEETS A ESTABLISHING PEAK POSITION AND HPCV TARGET RANGES Flow Check Fluorospheres Lot Number Expiration Date Flow Check 675 Fluorospheres Lot Number __ Expiration Date Flow Check 770 Fluorospheres Lot Number Expiration Date FS FL1 FL2 FL3 FL4 FL5 Red laser 488 Run HPCV Tech Date CO CO NW oo on Aw po Cytometer Serial No Greann Laboratory COULTER Cytomics FC 500 MPL 7579027B PN 177579BD A 13 LOG SHEETS A 14 PN 177579BD B 1 B 2 B 3 PN 177579BD USING MXP SOFTWARE Bo IMPORTANT If a laboratory wants both MXP and CXP Analysis software installed on an FC 500 MPL consider the following e CXP Analysis software must always be installed before the MXP software is installed If CXP Analysis software is installed on a Workstation with a previously installed version of MXP software lockups may be observed in the MXP software when executing the AutoSetup Wizard e CXP Analysis software must always be installed in the CXP directory not the MXP directory
16. If nothing happens or an Account Disabled message appears when you log in ask your System Administrator to unlock your User ID If you do not have a User ID and Password see your System Administrator The User ID Administrator and __ Admin are for the System Administrator s use only You cannot log into the standard MXP software with the Administrator User ID If you have the 21 CFR Part 11 option the System Administrator can use the File gt Log In feature to log into the MXP software but only after an automatic logout timeout or a user requested software lock Note If a red X appears next to your User ID lE techcomm notify the System Administrator Instructions are contained in the System Administrators Guide PN 177579BD PN 177579BD USING MXP SOFTWARE MULTI USER SIGN ON Log Into and Log Out of DaCS Software If you have the 21 CFR Part 11 option when you log into MXP software no additional actions are needed to be automatically logged into the DaCS software when data is exported to MS Excel When you are logged into MXP software the MXP software User IDs are also used by the DaCS software When you log out of MXP software you are automatically logged out of the DaCS software User ID This lists all the User IDs that are allowed access to the system User IDs are set up by the System Administrator If your name is not included see your System Administrator Logged In Indicates whether or not the Users
17. PN 177579BD PN 177579BD Data Source Legend Choose the File and Gate to be used for the Legend plot USING MXP SOFTWARE B PLOTS MENU File Legend Plot Properties Browse Button Gate See also Legend Data Source Legend Legend Options Tab Legend FCS Tab Format Plot Legend Options Tab The Options tab displays various checkboxes that may be enabled as appropriate Show Phenotype Logic Columns J Legend Plot Properties Gated Total Number Absolute Count See also Legend Data Source Legend Legend FCS Tab Format Plot B 117 USING MXP SOFTWARE PLOTS MENU B 118 Show Phenotype Logic Columns Displays the logical combination of Color Gates for each population Available in Color Blend Mode only Gated Total number of events in region as a percentage of events in a gated display Total Total number of events in region as a percentage of total events in the file Number Total number of events in a Region Absolute Count This displays the absolute count of a population based on the calibration factor CAL Factor and the number of particles within a defined CAL region Legend FCS Tab Shows FCS information Enable this xj checkbox to allow the display of sample DataSouce Options FCS information stores as part of the FCS header M Show FCS Information Show FCS Keyword Enable this checkbox to eee kea Eee SCYT XL P1008 DATATYPE
18. Redo reverses the last Undo operation If a new operation is performed after a series of Edit Redo operations the Edit Redo list is cleared and all previous Undo actions are lost The Edit Redo list is cleared after each change of plot focus Edit Cut deletes the highlighted items from the display placing them within the Clipboard These items can be recalled either by using the Copy Paste or Paste Special options within the Edit menu If you Edit Cut another item before the first item is Pasted this first item is overwritten See also Copy Paste Paste Special Copy PN 177579BD To Copy Statistics Regions or Plot Images onto the Windows Clipboard O Edit Copy This allows for Pasting these items into other areas of MXP software or to other Windows Applications such as MS PowerPoint Word Excel and so on The exact nature of what can be pasted depends upon the target application and can be controlled or modified using the target applications Edit Paste Special command See also Cut Paste Paste Special B 59 B USING MXP SOFTWARE EDIT MENU Paste After copying into memory MXP software allows the pasting of a number of different items The default paste result depends upon what you are trying to paste e Ifthe target is a FlowPAGE the Plot Image and Statistics are Pasted 1 Select the Plot of interest to be copied 2 C the Plot and hold down the left mouse button The
19. AutoSetup defaults 3 7 3 8 definition GLOSSARY 4 display in acquisition manager B 65 file extension 3 6 generic defaults 3 8 listmode compensation extension 3 6 new B 13 Parameter Mismatch when loading listmode files B 26 B 28 parameters Cytometer Control setup B 168 data source B 101 names in acquisition manager B 67 x and y data source B 96 passwords change 21 CFR Part 11 B 175 related to 21 CFR Part 11 C 1 signing on to MXP software B 3 paste edit menu B 60 paste special edit menu B 61 pause on error autogating failure B 55 PC7 definition GLOSSARY 5 PDF file extension 3 6 file name extension LMD file name B 47 format 3 5 performance specifications carryover 3 11 precision for surface markers 3 11 resolution 3 11 sensitivity 3 12 stability 3 12 throughput 3 11 See also specifications p8 definition GLOSSARY 5 PGE file extension 3 6 photodiodes FS and SS sensors 3 3 photo multiplier tube See PMT INDEX 16 physical specifications instrument 3 1 pickup lens spacial filter assembly definition GLOSSARY 5 picogram definition GLOSSARY 5 plate new B 14 plate data display file names 3 6 plate definition file definition GLOSSARY 5 plate display appears at startup B 1 Plate ID acquisition manager B 68 plate wizard definition GLOSSARY 5 description B 14 plot backgrounds changing to black B 52 plot display baseline offset B 53 auto B 53 off B 53 on B 53 defaul
20. B USING MXP SOFTWARE ANALYSIS MENU Editing and Moving a Region Name To edit or move the Region name 1 Double click on or near the first letter of the Region name to activate the Region Name dialog box 2 To edit the Region name type in the desired changes if any click Edit Region Name E x 3 To move the Region Name mouse over the region to get the hh cursor If the region name has multiple characters mouse over the first character to get the Hh cursor 4 D hold and drag to move the region name 5 D to fix the location of the region name B 140 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Editing a Quadrant Region Name 1 O Analysis gt gt View Modify Regions O H1 2 J on the region name pause and 0 D on the region name again to enter text mode 3 Type the new name and Loo Deleting Regions There are two methods used to delete degions From Within a Plot and Using the Delete Button in the View Modify Regions Dialog Box Note If a plot is deleted its regions are still retained From Within a Plot 1 the Region The Region is now active and handles appear 2 Press Delete The Region is now deleted Any Gates and statistics that were associated with this Region are also deleted PN 177579BD B 141 USING MXP SOFTWARE ANALYSIS MENU Using the Delete Button in the View Modify Regions Dialog Box OBI 2 the Regio
21. B 69 CAL region B 153 calculate results B 153 calibrating absolute counts B 152 carry cytosettings B 67 regions from previous tube B 66 regions from protocol B 66 carryover performance specifications 3 11 cascade Windows B 179 category installation 3 1 cell type user info B 51 channel definition GLOSSARY 1 check in file B 71 cleaning agent definition GLOSSARY 1 description 1 4 cleanse Cytometer B 173 Cleanse Mode use of cleaning agent 1 4 click definition GLOSSARY 1 clip channels scaling histogram plot B 105 close all Windows B 181 CMP compensation file extension 3 6 CMP file import settings B 42 PN 177579BD coefficient of variation CV See CV collimate definition GLOSSARY 2 color precedence advanced B 131 precedence automatic B 54 color blend mode B 127 modify color blend B 128 color compensation definition GLOSSARY 2 QC material used to adjust settings 1 5 specifications 3 4 See also compensation color dot plot coloring mode B 55 copy plot data B 96 copy plot image B 95 creating B 95 data source B 96 events B 97 apply to all plots B 98 no of events to show B 98 font B 99 format plot B 96 labeling B 98 show axis labels B 99 show axis tick marks B 99 show plot title on output B 99 x and y axis labels B 99 resolution B 98 compensation clear all Cytometer Control B 170 Cytometer Control B 170 file extension 3 6 listmode B 92 matr
22. INDEX INDEX 24 PN 177579BD BECKMAN COULTER INC CUSTOMER END USER LICENSE AGREEMENT This Product contains software that is owned by Beckman Coulter Inc or its suppliers and is protected by United States and international copyright laws and international trade provisions You must treat the software contained in this Product like any other copyrighted material This license and your right to use the Product terminate automatically if you violate any part of this agreement This is a license agreement and not an agreement for sale Beckman Coulter hereby licenses this Software to you under the following terms and conditions You May 1 Use this software in the computer supplied to you by Beckman Coulter 2 Maintain one copy of this software for backup purposes the backup copy shall be supplied by Beckman Coulter 3 After written notification to Beckman Coulter transfer the entire Product to another person or entity provided you retain no copies of the Product software and the transferee agrees to the terms of this license agreement You May Not 1 Use copy or transfer copies of this Software except as provided in this license agreement 2 Alter merge modify or adapt this Software in any way including disassembling or decompiling 3 Loan rent lease or sublicense this Software or any copy Limited Warranty Beckman Coulter warrants that the software will substantially conform to the published specifications for
23. Regions After Regions have been created use this option to select or hide which Regions you wish to view on a plot Existing Regions are listed in the Select Regions list box The highlighted Regions correspond to the Regions already displayed in the current plot window Only Regions sharing the same parameters and Gates as the current plot can be displayed Display Regions 1 w the region names you want to OK display on the plot If Automatic Gate Maintenance is enabled gating regions do NOT appear in the list of available regions Cancel Select All Help sikh Cie Dot Plot Events Use this to specify the percentage of events to view This option significantly increases the speed at which dot plots refresh if large data files are being analyzed For an event interval less than 100 data is displayed uniformly from within the whole of the data file that is if 10 of events are selected then event numbers 1 11 21 31 and so on are displayed if they meet the Gating criteria for the plot The default value for the number of events to display can be set in the File gt Workspace Preferences and selecting the Plot Display tab No of Events to Show x Apply to All Plots Labeling Font Stop and Save Data Source Events Resolution Total No of Events 21374 age Total Events 100 No of Events to Show 21374 i i I Apply to all plots Cancel Help See also Workspace Pr
24. This allows you to select the style and which Toolbars are displayed Toolbars xi Toolbar Name Toolbars Commands Show ToolTips Toolbars Cool Look IV Show Tooltips New Large Buttons Bey Beset N J Large Buttons ew Acquisition Manager Tools Reset Toolbar name Fie Cancel Help If a customized Toolbar has been created and this is highlighted Dete is available to allow the deletion of that Toolbar See also Toolbars Customize Command Toolbars Select which Toolbars you wish to be displayed Any customized Toolbars are listed Toolbar Name Allows you to modify the name of any customized Toolbar It is grayed out for the standard Toolbars Show ToolTips Show Fly over help when the mouse is pointed to the Icon Cool Look This changes the style to a cool look Large Buttons Changes the Icon buttons to a Large size New Displays a dialog box to enter your customized name of your new Toolbar Reset Resets the Toolbar to the default values PN 177579BD USING MXP SOFTWARE VIEW MENU Toolbars Customize Command This allows you to customize the Toolbars by dragging the button to the desired position within any Toolbar Categories x Buttons Toolbars Commands Description Categories r Buttons Acquisition Mana Flowpage Batch R Select a category then click a button to see its description Drag the button to any toolbar Description
25. no statistics have been selected then the default statistics are displayed FCS options are either generated automatically by MXP software or obtained from sample information entered by the User Desired items can be selected and exported during the publishing of an Excel Spreadsheet The selected FCS information is only published and does not print with the plots within MXP software When in Color Blend Mode only the color blend color statistics from the blended colors are displayed in the Legend plot Use to view Region statistics sialic Type x Numb er Statistic Type r Report Options Total Current pags IV Mean Calculation Method Gated Comment Percentile Mean FCS Information CY BTIM 15 50 32 Median HPCV Mode Cells pL Min and Max Cancel Advanced Help Current File Name Mean Calculation Method FCS Information Advanced Statistics Configuration For more information concerning the Publication of Results see Publish Results Now PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Statistic Type There are the different types of statistics that can be utilized Any number of these can be selected and the relevant statistic appears in the box beneath the plot Current File Name This option allows you to include the data file name when the results are published to an Excel Spreadsheet This information is not displayed within the Statistics Window Mean Calculation Method This exp
26. options B 93 Parameter Mismatch B 26 B 28 playback B 90 list comp setup B 92 options B 93 QuickCOMP B 153 QuickCOMP matrix B 154 single file mode B 91 single file mode B 91 specifications 3 6 Listmode playback tool definition GLOSSARY 4 LiveGate definition GLOSSARY 4 LMD definition GLOSSARY 4 INDEX 13 INDEX LMD file name date B 46 display in acquisition manager B 69 file name extension B 47 PDF file name extension B 47 append PDF file name B 47 run number B 46 sample ID 1 2 3 4 B 46 tag number B 47 time B 46 user ID B 46 well loc B 46 well number B 45 workspace preferences B 45 LMD QuickCOMP B 153 matrix B 154 load cytosettings B 67 regions from current protocol B 66 lock out automatic 21 CFR Part 11 C 2 definition GLOSSARY 4 manual 21 CFR Part 11 B 175 C 2 MXP software B 175 log in DaCs software B 3 MXP software B 57 procedure B 58 system administrator B 57 log out automatic 21 CFR Part 11 C 2 DaCs software B 3 MXP software B 58 log scale density plots B 52 log sheets action log A 1 daily log for instrument standardization A 1 daily log for instrument verification of alignment and fluidics A 1 establishing fluorescence and or light scatter target ranges A 1 establishing HV total gain ranges A 1 reagent log A 1 logarithmic amplification definition GLOSSARY 4 logged in DaCs software B 3 signing on to MXP software B 3 INDEX 14 macros See MPL
27. 1 MULTI USER SIGN ON B 1 Plate Display B 1 Startup Wizard B 2 Log Into and Log Out of DaCS Software B 3 User ID B 3 Logged In B 3 Password B 3 Admin Button B 3 Finish Button B 3 TOOLBAR BUTTONS B 4 FILE OPTIONS TOOLBAR B 5 PLOT OPTIONS TOOLBAR B 5 REGIONS OPTIONS TOOLBAR B 6 GATE COLOR STATS AND HELP TOOLBAR B 6 FLOWPAGE TOOLBAR B 7 ACQUISITION MANAGER TOOLBAR B 7 AUTOMATOR TOOLBAR B 8 CYTOMETER TOOLBAR B 9 LISTMODE PLAYBACK TOOLBAR B 10 CONTENTS vii CONTENTS B14 viii FILE MENU B 11 New Protocol B 11 Creating a New Acquisition Protocol B 12 Creating a Protocol for Analysis B 12 New Panel B 13 New Plate B 14 Sample Plate Analysis with MPL_DaCSAnalysis Add In or MPLAnalysis Add In B 15 Open Listmode File B 23 Open Listmode File Procedure With 21 CFR Part 11 Option Installed B 23 Open Listmode File Procedure Standard MXP Software Installed B 27 Save Listmode File As B 28 Save Listmode File As Procedure With 21 CFR Part 11 Option Installed B 29 File Name B 29 Save Listmode File As Procedure Standard MXP Software Installed B 30 File Name B 30 Open Histogram B 30 Save All FCS Format Files B 31 Save Histogram File As B 32 Save Histogram File As Procedure With the 21 CFR Part 11 Option Installed B 32 Save Histogram File As Procedure Standard MXP Software Installed B 33 Open Protocol B 34 Open Protocol Procedure With the 21 CFR Part 11 Option Installed B 34
28. 1 start B 171 stop B 172 toolbar B 9 Cytometer Control acquisition setup B 165 baseline offset B 167 discriminator B 168 dots B 168 drive space B 165 duration B 165 live gate B 165 max events B 165 red laser button B 168 setup mode B 166 button B 164 compensation B 170 clear all B 170 matrix B 170 detectors B 169 AUX parameter source B 170 gain B 170 names B 169 volts B 170 parameter setup B 168 QuickCOMP B 167 QuickSET B 167 cytometry See flow cytometry PN 177579BD Cytosettings definition GLOSSARY 2 cytosettings carry B 67 display state in acquisition manager B 67 get cytosettings from LMD file B 171 get cytosettings from protocol B 171 load B 67 Cyto Trol control cells description 1 5 See controls D DaCs software 21 CFR Part 11 compliance C 4 audit trails C 5 description 1 3 electronic signatures C 6 Excel worksheets C 5 log in B 3 C 4 log out B 3 C 5 MPL DaCs Analysis Add In B 15 setting up user IDs C 6 user s manual C 5 daily log for instrument standardization A 1 for instrument verification of alignment and fluidics A 1 data acquisition throughput performance specifications 3 11 data rate status bar B 174 data sheets material safety how to order 1 5 data source apply gate to all plots B 96 color dot plot B 96 density plot B 107 file B 96 gate B 96 histogram plot B 101 legend plot B 117 parameter B 101 PRISM plot B 111 regions B
29. 115 editing dual parameter PRISM dividers B 144 single parameter PRISM divider B 146 editing dividers B 114 explained B 112 phenotypes B 112 removing dividers B 115 scaling B 111 view modify B 147 PRO extension for protocol file 3 6 project user info B 51 protocols AutoSetup defaults 3 7 3 8 axis calibration keep the same when modify 3 8 compensation 3 7 3 8 creating analysis B 12 creating new acquisition protocol B 12 definition GLOSSARY 5 display in acquisition manager B 65 file extension 3 6 Flow Set 3 7 3 8 generic defaults 3 8 new B 11 open B 34 runtime definition GLOSSARY 6 save B 37 save as B 38 settings 3 7 3 8 verification 3 7 3 8 publish histogram data B 84 publish data to Excel with an Analysis Add In B 15 publish data to MS Excel B 56 publish data to text file B 56 results B 84 INDEX 18 switch to MS Excel on publish B 57 workspace preferences B 56 Q QC materials listing 1 5 quadrant region creating B 135 editing re naming B 141 properties B 135 quality control QC definition GLOSSARY 5 materials 1 5 materials use and function 1 5 QuickCOMP definition GLOSSARY 5 enabling for analysis B 167 listmode playback B 154 LMD B 153 QuickCOMP 2 kit description 1 5 See also controls QuickCOMP 4 kit description 1 5 See also controls QuickSET definition GLOSSARY 5 enabling B 167 R RD1 definition GLOSSARY 5 reagent log A 1 reagents
30. 116 Legend Use this option to view the population colors population names and FCS Header sample information Note The Amplifier Gain field is no longer available at this screen if you adjusted the compensation of a Listmode file using the LMD QuickCOMP tool The original stored file remains unchanged and contains the Amplifier Gain information 1 Plots Legend Plot to create a legend plot When a Plot is displayed click the right mouse button within the plot to display further menu options Every time the population names or colors E lungated Legend are edited MXP software updates the Legend Color Stat Gated Total Number Window If MXP software is in Dot Plot Mode Color Blend mode Legend will display the names colors and associated statistics of the color Gate combinations 2 If MXP software is in Dot Plot Mode Color Precedence mode Legend displays the names colors and associated statistics of the Gates with the highest precedence at the top Note When analyzing multiple files using Color Precedence the Legend plot shows all gates from all plots regardless of precedence and file To change this so that the Legend shows only those gates and plots associated with a given file use the Advanced Precedence tab on the Color Precedence edit dialog See also Data Source Legend Legend Options Tab Legend FCS Tab Color Blend Mode Introduction Modify Color Precedence
31. 13 w ta 200 15 150 16 1 100 18 50 19 o z 1 2 3 4 s 6 7 8 s 10 22 Well Number 23 24 25 Threshold S6OI6 26 27 28 29 Fie Gae XParam_ Y Param_Fiename Mean Calc jon Variable Value Well Concentration 70 101 5000 bead BFL Lo 01 S000 L0G L00 S000 Caibrator R 1 _JO 1 u10 Number aaa 1 71 02 2500 dbead3 BAFL2 Lor 022500 LOG LOG 2500 Calirator R1 D_ 2 L 10 Number 330 72 03 1250 dbead3 BqFL2 Le fo3_1250 LOG L 1250 ColbretorR 1 D 4 L 10 Number 263 3 73 04625 Clbead3 BdFL2 Log fo4 625 C LOG LOG 625 Caibrator R 1 0_ 8 L 10 Number 333 4 74 05_312 Clbead3 BAFL2 Log 05 312 C1LOG L 312 Caibrator R_1 b 16 u 10 Number 5 75 06156 Clbead3 BA FL Log 06 156 C1LOG L 156 Caibrator R_1 D_ 32 1 10 Number 323 5 76 07 _080 C bead3 Bd FL Log 07 080 C1LOG L 180 Calrator R_4 lb 64 u 10 Number 353 7 77 08 040 C bead3 BAFL2 Log 05 040 CLLOG L 40 Caibrator R_1 b 126 L 10 Number E 78 09 020 C bead3 BdFL Log 09 020 C1LOG L 20 Caiorator R 1 D_ 256 ut Number E 79 10_neg Sjbead3 B FL2 Log 10 neg SJLOG LOG neg Specimen R 1 jundefinediL10 Number 327 10 22 If calibrators were run a a E E E E E z Cal_IL 10X Median Separate sheets are created for Ee each Mean Median or Mode A ee statistic exported He a e The calibration curve is a third Ha j or
32. 173 Status Bar B 173 Total Events B 174 Elapsed Time B 174 Data Rate B 174 Flow Rate B 174 Cytometer Status B 174 Windows Status Bar B 174 X and Y Coordinates B 175 21 CFR 11 MENU B 175 Lock Software B 175 Change Password B 175 Procedure B 176 Sign File Version B 177 Procedure B 177 WINDOW MENU B 179 Cascade B 179 Tile B 179 Tile Special B 180 Arrange Icons B 181 Force Square B 181 Close All B 181 21 CFR PART 11 OPTION COMPLIANCE INFORMATION C 1 OVERVIEW C 1 INTEGRITY OF ELECTRONIC DATA C 1 Passwords C 1 Automatic and Manual Lock Out C 2 MXP Software Files C 3 Windows Operating System C 3 Third Party Software Applications C 3 CONTENTS xvii CONTENTS had kot LO C 6 ARCHIVING AND RESTORING ARCHIVED DATA C 4 AUDIT TRAILS C 4 ELECTRONIC SIGNATURES C 4 DaCS SOFTWARE C 4 User s Manual C 5 Audit Trails DaCS Software C 5 Electronic Signatures DaCS Software C 6 Setting Up User IDs C 6 MXP Software User IDs C 6 DaCs Users For Use Outside MXP Software C 6 GLOSSARY INDEX BECKMAN COULTER INC CUSTOMER END USER LICENSE AGREEMENT TRADEMARKS xviii PN 177579BD CONTENTS ILLUSTRATIONS Ll a ce 2 3 24 PN 177579BD Cytomics FC 500 MPL Flow Cytometry System 1 1 Power and Signal Cable Connections with Voltage Specific Power Supply 2 3 Power and Signal Cable Connections with Universal Power Supply 2 4 Waste and Pneumatic Tubing Connecti
33. 1L Settings PRO 8 AS SC 1L Verify PR a A5 2C Flow 5et PRO a AS 4C 2L Settings PRO a AS SC 2L Settings a AS 2C Neg Control PRO a AS 4C 2L Flow Set PRO a AS SC 2L Flow Set F AS 2c verify PRO a AS 4C 2L verify PRO AS SC 2L verify PRC C File gt Save Protocol As 2 aJ AS 4C IL VeriFy PRO a AS 5C 1L Settings PRO 8 45 FL1 Comp PRO fe AS 4C 1L Flow Set PRO se AS SC 1L Flow Set PRO 8 AS FL2 Comp PRO In the Save Protocol As dialog box gt File name AS 5C 2L Verify PRO 5 the desired folder from the list or by typing the path in the File name text Se ot020l Fes pro z Canos Y LA box B 39 B USING MXP SOFTWARE FILE MENU 3 In the File name text box type the desired File Name MXP software gives the default file extension PRO The PRO extension is only a suggested extension any standard DOS characters can be used for the full File Name You are urged to keep the extension consistent for easy file searches Save Listmode Compensation As Use this feature to save the currently loaded compensation settings into a CMP file These compensation files are most often used with the Listmode playback feature View FCS Header Information O xi File gt View FCS Header File C MXP Users SharleneS LMD 5C 2L AS Verify Information to view the FCS header information on the current file Note The Amplifier Gain field is no longer available
34. 2004 17 17 08 Signature Certified Signed by Mr Anderson Signed on 06 Oct 2004 17 17 23 Signature Authorized Signed by Mr Anderson Signed on 06 Oct 2004 17 17 38 z B 35 B USING MXP SOFTWARE FILE MENU 7 Atany time you can D to hide the Checked In Versions portion of the screen 8 To open the file you want highlight the file name and either D to open the most recent version of the selected file OR e Highlight a specific version of the file at the bottom of the screen and J to open that version Note Double clicking with the mouse on the file name or version closes the dialog box and opens the file Note If you selected an archived file that has not been restored a message appears stating the archive location of the file Supply the file location to your System Administrator and request the file be restored B 36 PN 177579BD USING MXP SOFTWARE FILE MENU Open Protocol Procedure Standard MXP Software Installed Use this procedure if you have the standard MXP software installed without the 21 CFR Part 11 option Look in SY AcquisitionProtocol EE EB AS 4C 2L Verify PRO aas FL3 Comp PRO AS 5C 1L Settings PRO sa AS FL4 Comp PRO AS 5C 1L Flow Set PRO 8 AS FLS Comp PRO AS 5C 1L Verify PRO ja Cleanse Bleach PRO AS SC 2L Settings PRO 38 Cleanse Water PRO AS 5C 2L Flow Set PRO ajo 1L Flow Check PRO AS SC 2L Verify PRO Qc w Check PRO 1 File gt Open Protocol
35. 97 x and y parameter B 96 data storage drives 3 4 specifications 3 4 PN 177579BD INDEX date LMD file name B 46 db definition GLOSSARY 2 dc definition GLOSSARY 2 default of events on dot plot B 53 3d plot mode B 53 AutoSetup protocols and panels 3 7 3 8 definition GLOSSARY 2 generic protocols and panels 3 8 log scaling acquisition options B 49 overlay mode B 53 overlay plot background color B 53 plot size B 53 definitions GLOSSARY 1 delete edit menu B 61 FlowPAGE objects B 156 histogram freeze frame overlay B 104 PRISM plot dividers B 115 regions B 141 delivery inspection instrument 2 1 density density plot automatic maximum threshold B 108 density plot level color B 108 density plot logarithmic threshold calculation B 108 density plot maximum density level B 108 plot resolution B 109 density plot automatic maximum threshold B 108 configuration B 107 creating B 106 data source B 107 density level color B 108 labeling B 109 log scale B 52 logarithmic threshold calculation B 108 maximum density level B 108 resolution B 109 smooth B 109 use gate color B 109 use single color range B 109 detectors Cytometer Control B 169 AUX parameter source B 170 INDEX 7 INDEX detector gain B 170 detector names B 169 detector volts B 170 dimensions computer 3 1 Cytometer 3 1 instrument 3 1 Power Supply 3 1 Workstation 3 1 DiOC5 3 definition GLOSSARY 2 disable
36. B 122 advanced precedence B 131 automatic B 54 blend color B 128 boolean gating B 125 color blend mode B 127 gate and logical expression B 122 gate color B 124 gate logic B 125 gate name B 128 modify color blend B 128 modify color precedence B 130 operands B 123 data source apply to all plots B 96 maintenance automatic B 54 maximum per protocol 3 5 options toolbar B 6 plot data source B 96 gating definition GLOSSARY 3 workspace preferences B 54 GB definition GLOSSARY 3 ground path requirement 2 2 H hardware options 1 3 heat dissipation 2 2 help toolbar B 6 help toolbar B 6 HeNe laser width 3 1 See also laser high voltages amplification 3 3 definition GLOSSARY 3 histogram definition GLOSSARY 3 INDEX 11 INDEX file open B 30 file save as B 32 PRISM definition GLOSSARY 5 resolution 3 5 histogram plot configuration B 101 creating B 100 data source B 101 freeze frame B 102 creating B 103 deleting overlay B 104 introduction B 102 labeling B 105 scaling B 105 automatic maximum value B 105 clip channels B 105 set maximum B 105 smooth B 102 use gate color B 102 HPCV definition GLOSSARY 3 explanation B 150 specification for fluorescence 3 11 specification for forward scatter 3 11 humidity allowance 2 2 hydrodynamic focusing definition GLOSSARY 3 icons See buttons toolbars idle mode Cytometer B 173 ignore errors autogating failure B 55 Immun
37. B 54 gate creation B 54 gate maintenance B 54 lock out 21 CFR Part 11 C 2 log out 21 CFR Part 11 C 2 maximum value scaling histogram plot B 105 region name positioning B 133 setup target region B 136 AutoMATOR add blank B 88 add files B 88 introduction B 85 INDEX 3 INDEX output options B 89 panelize B 88 path details B 89 remove files B 88 re ordering files B 87 restore analysis files on startup B 89 setup B 85 status bar B 90 toolbar B 8 AutoSetup default panels and protocols 3 7 3 8 initial protocol 3 7 modified protocols must have same axis calibration 3 8 panels 3 7 3 8 protocols 3 7 3 8 target region B 136 AutoSetup Wizard definition GLOSSARY 1 AUX parameter signal definition GLOSSARY 1 source Cytometer B 170 axis calibration keep same for modified protocols 3 8 background count definition GLOSSARY 1 baseline offset B 53 auto B 53 off B 53 on B 53 batch AuttoMATOR See AuttoMATOR BCAG P database definition GLOSSARY 1 beam shaping optics 3 2 blue laser See Argon laser boolean gating B 125 gate logic B 125 bottle waste See waste container BP filters definition GLOSSARY 1 Btus required for operating system 2 2 INDEX 4 buttons definition GLOSSARY 1 See also toolbar C cable connections location 2 3 power 2 3 signal 2 3 Cal factor B 152 absolute calibration particles B 153 definition GLOSSARY 1 in acquisition manager
38. B 98 show axis labels B 99 show axis tick marks B 99 show plot title on output B 99 x and y axis labels B 99 density plot B 109 histogram plot B 105 laser beam spot size Argon 3 2 beam spot size solid state 3 2 definition GLOSSARY 4 disable red B 168 dual laser filter block 1 3 dual laser filter kit 1 3 operating wavelengths 3 2 second red laser option 1 3 shutter for second laser 1 3 single laser filter block 1 3 single laser filter kit 1 3 solid state description 1 3 specifications 3 2 types 3 2 width HeNe 3 1 laser beam spot size Argon 3 2 solid state 3 2 laser width HeNe 3 1 layout of the instrument 1 1 legend plot creating B 116 data source B 117 FCS tab B 118 PN 177579BD INDEX options B 117 gated B 118 total B 118 number B 118 show phenotype logic B 118 light scatter standardize intensity 1 5 linear amplification See gain linear region creating B 136 interactive information B 136 properties B 136 automatic name positioning B 133 lower and upper limit B 136 minimum count B 136 positives analysis B 136 prime B 136 region name B 136 target region for automatic setup B 136 See also regions linearity monitor instrument 1 5 linked marker creating B 137 listmode batch replay B 85 compensation B 92 compensation panel extension 3 6 data definition GLOSSARY 4 FCS file save as B 28 file open B 23 maximum file size 3 6 maximum parameters 3 6
39. Comments OR e Type in a comment in the Comments for this version field Note Once a comment is entered the version number automatically appears in the Saving As Version field and the button is active 4 W to save the file MXP software creates a file with the name and with the version and comment if applicable that you specified and stores it in the drive and directory you specified in FCS format File Name A listmode file name can have up to 200 alphanumeric characters including the filepath However the following characters cannot be included forward slash backslash greater than sign gt less than sign lt asterisk comma question mark quotation mark pipe symbol colon or apostrophe Leading Trailing and multiple spaces are not allowed B 29 B USING MXP SOFTWARE FILE MENU Save Listmode File As Procedure Standard MXP Software Installed Use this procedure if you have the standard MXP software installed without the 21 CFR Part 11 option Use this Save As feature if at any time you decide to save your listmode file under a different File Name 1 D File gt Save Listmode File As 2 Specify the name of the file zix Save in Human Cytokine tdf 20030612162217 z e m 1 vandy 00001059 2003 06 12 1620 622 LMD 02_Cytokine p 2500 Calibrator R_1 D_ 2 vandv 00001060 2003 06 12 1622 623 LMD 03_Cytokine p 1250 Calibrator R_1 D_ 4 vandv 00
40. Fs Lin FS Lin z ss Lin SS Lin 7 Regions I Apply gate to all plots Density Level Configuration Use this option to specify the Density levels and other configuration preferences Density Level Colors x Automatic Maximum Threshold e o e Maximum Density Level Data Source Density Resolution Logarithmic Threshold Calculation Density Level Colors Smooth Density Plot Levei W j Use Gate Color level2 z v Logarithmic Threshold Calculation Use Single Color Range y 8 8 beclls E I Smooth Density Plot Level 4 I Use Gate Color Nereis E J Use Single Color Range Use Defaults Cancel Help PN 177579BD B 107 USING MXP SOFTWARE PLOTS MENU B 108 Density Level Colors This Plot may have up to 5 different ISO count levels each of which is represented by a color Choosing the appropriate button may change the default color for each level 1 D the Level 1 button to view the Color dialog box 2 Select the desired color that is to be allocated to Level 1 3 Continue as above until all desired changes have been made O to accept all changes w or press to exit the dialog box without saving the changes Automatic Maximum Threshold This option allows automatic configuration of the maximum value that is used to calculate the Density Color Levels If this option is not selected you may manually enter the value which is used to calculate the Density Color Levels Maximum Density
41. Gate has been deleted any plot that used the deleted Gate is reset to Ungated Allows you to resize an already created Region PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Status Bar Linear The status bar normally gives a brief description of selected menu items When drawing or modifying Regions the status bar divides into three areas of information These update as a region is modified e The status bar shows Channel for X parameter e The total integral count of the region e The percentage of events currently within the region X Coordinate The X Axis channel value is the coordinate of the cursor position given by a scaling factor This scaling factor depends on whether the Histogram parameter is using linear or logarithmic amplification and whether you have calibrated that parameter or not If the parameter is linear and uncalibrated the X channel varies between 0 255 or 0 1024 depending on the data source If the parameter is logarithmic and uncalibrated the value varies between 0 1024 1024 and span four decades For most other manufacturers the data varies between 1 10 000 spanning four decades Integral When you create a Region the status bar reflects the integral values given along the scale for that parameter at the cursor position That is before the first marker is set If the cursor is on channel 10 the integral shows the count value of the populations at channel 10 After first marker is set an
42. If CXP Analysis software is installed in the MXP directory the CXP Analysis Help files replace the MXP Help files so the MXP Help files are no longer available for access on the FC 500 MPL WELCOME TO MXP SOFTWARE Welcome to MXP software This exciting new software designed in conjunction with customer feedback is simple fast and reliable allowing you to obtain the best from your Cytometer CHECK FOR BCA amp P DATABASE ICON Before you launch the MXP software check that this icon ab appears in the Windows Qo DIN ajz 12 26 PM system tray This indicates the BCA amp P database is online MULTI USER SIGN ON Plate Display The Plate Display appears with a default undefined 96 well plate configuration Once a user logs on the Plate Display window shows the last plate configuration defined by that user Note When anyone logs in with a new User ID Not a valid worklist file appears when the MXP software is launched Until a valid worklist is saved for that User ID this message appears after log in Untitled Plate Disp Ioj xj OCCOCCGCCCoe COCCCGCOCOCEeoe oOGCcoceecoeoceso OCOLO COoeocoocoGoocoo OCOCOCOCGOCOCOe COSCCCOCOCee COGCocoeeooes B 1 USING MXP SOFTWARE MULTI USER SIGN ON Startup Wizard User ID MXP Cytometer Startup Wizard S x Logged In Password UserID Logged In i SAME Administrator No Admin Button SH Sharlene No Finish Button SM Dine No 7 SH Theresa No GERN MXP Sof
43. Level Configure the Density Plot to manually enter the value that is used to calculate the Density Color Levels This option is only active when the Automatic Maximum Threshold button is cleared Logarithmic Threshold Calculation Density color thresholds may be calculated in one of two ways e Linear threshold calculation option toggled off The maximum value is used to determine the threshold value for each color The actual threshold value is calculated as follows Value Level No x Maximum In Plot 5 1 e Logarithmic threshold calculation option toggled on The maximum value is used to determine the threshold value for each color The actual threshold value is calculated as follows Value 10 Level No x log10 Maximum In Plot 5 1 PN 177579BD PN 177579BD USING MXP SOFTWARE PLOTS MENU Smooth Density Plot Choose this option to smooth the Density plot Clear it to display raw unsmoothed data Use Gate Color Creates a single color density range based on the currently assigned gate color Use Single Color Range Creates a range of density level colors based on the selected Level 1 and Level 5 colors Density Plot Resolution Set the number of channels for the X and Y Axis of the Plot The available options allow for 64 128 256 E and 512 channels to be displayed on the a e ee axis Data Source Density Resolution Resolution of plot A higher resolution gives an improved graphic image whereas a
44. Picture FlowPAGE allows the import of a Windows Bitmap file extension bmp 1 Use MS Paint or other graphics editor to create a bitmap file to be placed on a FlowPAGE and save it as a file W Insert gt gt Other FlowPAGE Object and W the Picture option 2 Select the desired bitmap file Open HAE Look in E Sharlene 7 t iad Fa C AcquisitionProtocol C Panel i 7 i AnalysisProtocol PDF a the desired directory in the we cas Look in list box HTML E worklist b the type of file in the Files of n ile name es type list box i Files of type Bitmap Files bmp dib gt Cancel WA c O the name of the bitmap file you want to insert Note Double click on the file name to close the dialog box and display the file 3 J the FlowPage location where you want to place the picture See also Inserting a Line Inserting a Rectangle Inserting a Text File PN 177579BD B 81 B USING MXP SOFTWARE INSERT MENU Inserting a Text File 1 Insert gt Other FlowPAGE Object and D the Text File option Note Use MS Notepad or other text editor to create text txt files 2 Select the desired text file Look in i admin h Al cl a D the desired directory in the Html Ci Resuts Look in list box Images C worklist b the type of file in the Files of type list box Filename Files of type ee Cancel p G W the name of the text file you want to insert Note Double
45. Plot Backgrounds This displays the plots with a black background instead of white This can be useful as it makes the viewing of smaller populations of cells much easier When you print drag or copy a plot to other software programs the background remains white Opaque Region Labels When black backgrounds are chosen the region labels are not visible since the text color is not automatically inverted This option displays an area around the annotation within black backgrounds making it visible Print Plot Frames This option can be used to show or hide frames on plots printed directly from the File Print menu and exported plot images If this is checked all frames are displayed around plots If it is clear frames are not printed or exported Print Dots Black Select this option to print all dots in black irrespective of their colors If it is clear dots are printed in various shades of gray or if using a color Printer the best matched colors available This option does not affect the printing of FlowPAGEs Show Overlay Legend Pane This feature is not available in MXP software Show Overlay Statistics Pane This feature is not available in MXP software Log Scale Density Plots Selecting this option displays any plot with a density mode for example density plot with log scaling of the density levels If unchecked linear scaling of the density levels is used These changes are only applied to plots created after this option h
46. a Protocol other than the current Protocol in memory J Cytometer gt Get Cytosettings From Protocol File and follow the instructions to open a protocol file in the heading Open Protocol Use the appropriate procedure depending on whether you have the standard MXP software or also have the 21 CFR Part 11 option installed on your system Get Cytosettings from LMD file The Get Cytosettings From LMD File option allows Cytometer Instrument Settings to be obtained from a listmode file D Cytometer gt Get Cytosettings From LMD File and follow the instructions to open a listmode file in the heading Open Listmode File Use the appropriate procedure depending on whether you have the standard MXP software or also have the 21 CFR Part 11 option installed on your system Start The Start button gt initiates acquisition of the current sample PN 177579BD B 171 B USING MXP SOFTWARE CYTOMETER MENU B 172 Stop The Stop button ends data acquisition Any Output options such as saving data calculating statistics printing reports and so on are then performed Abort Selecting the Abort button x displays the Abort screen This function is only available if Acquisition is running Note No Files are saved if a sample is Aborted Select the option required and O Loox e dae Cn x Abort the Panel Abort the Plate Do you want to Repeat the Well Abort the Well Repeat the Panel C Abort the Panel C Abort the Pl
47. and Dropped into the required position Panelize This sets the files into groups or panels based on the number of files open in the workspace The report icon displayed next to a file identifies the last file in each group Changes made to the Pause or Stop options for the first panel may be carried through all selected files by selecting Panelize If you attempt to run a test when the files are panelized and an incomplete panel is included Incomplete Panel appears giving you the option of going back to the AutoMATOR to amend the problem Load Queue Displays a dialog box to load a queue ALQ file Save Queue Displays a Save dialog box to save the queue to a file The files are saved in the LMD subdirectory under the file extension ALQ PN 177579BD Print USING MXP SOFTWARE TOOLS MENU Print a listing of the files in the queue Restore Analysis Files on Startup If the checkbox is enabled all the files displayed in the Analysis File list appear in the list the next time that you log in If the checkbox is not ticked the list is empty the next time you log in Show full pathname Shows the paths to the destination directories for the selected listmode files Output Options These options consist of several checkboxes found at the bottom of the AutoMATOR Setup dialog box Check the required output options to print or publish data at the completion of each sample The options are PN 177579BD Print plots as disp
48. any archived files that you need to access before proceeding Only the System Administrator can archive files and restore archived files To open a listmode file for the first time 1 W File Open Listmode File PN 177579BD B 23 USING MXP SOFTWARE FILE MENU 2 Select the desired listmode file Open LMD File Look in 5 color 2 laser autosetup tdf_20040930_115330 xi Ea 01_5c 2 as flow set Imd le 06_5c 21 as pe comp Imd I Show Archived Files if you be 02_5c 21 as fite comp Ind b 10_5c 2l as verify 8 19 3 45 4 md x G i Bad 03_5c 21 as pe comp Imd want to display archived files in Z 04 Se 2 as ecd comp ind Bar x le 05_5c 21 as ape comp Imd addition to the current listmode File name 05c 2l as flow set lmd Open files appears next to each ooro MAME z cea a J Open Runtime Protocol Rive archived file name I Show Archived Fies awar D the desired directory in the Look in list box The Trays directory has tdf files which contain multiple LMD files W the type of file in the Files of type list box C the name of the listmode file you want to view 3 W Open Runtime Protocol to replay the LMD into the plots in the runtime protocol If the open runtime protocol is not selected the LMD replays into the plots currently displayed on the workspace Note Listmode replay uses the runtime protocol Cal Factor unless you select a different Cal Factor on the Absolute Count Calibration dialo
49. are located in the C MXP admin directory Panel Name Flow Set Protocol Compensation Protocols Verification Protocol Settings Protocol AS 2C PNL AS 2C Flow Set Pro AS FL1 Comp Pro AS 2C Verify Pro AS 2C Settings Pro AS FL2 Comp Pro AS 3C 1L PNL AS 3C 1L Flow Set Pro AS FL1 Comp Pro AS 3C 1L Verify Pro AS 3C 1L Settings Pro AS FL2 Comp Pro AS FL4 Comp Pro AS 4C 1L PNL AS 4C 1L Flow Set Pro AS FL1 Comp Pro AS 4C 1L Verify Pro AS 4C 1L Settings Pro AS FL2 Comp Pro AS FL3 Comp Pro AS FL4 Comp Pro AS 4C 2L PNL AS 4C 2L Flow Set Pro AS FL1 Comp Pro AS 4C 2L Verify Pro AS 4C 2L Settings Pro AS FL2 Comp Pro AS FL3 Comp Pro AS FL4 Comp Pro AS 5C 1L PNL AS 5C 1L Flow Set Pro AS FL1 Comp Pro AS 5C 1L Verify Pro AS 5C 1L Settings Pro AS FL2 Comp Pro AS FL3 Comp Pro AS FL4 Comp Pro AS FL5 Comp Pro AS 5C 2L PNL AS 5C 2L Flow Set Pro AS FL1 Comp Pro AS 5C 2L Verify Pro AS 5C 2L Settings Pro AS FL2 Comp Pro AS FL3 Comp Pro AS FL4 Comp Pro AS FL5 Comp Pro The initial protocol in each panel which you use to adjust or confirm correct PMT High Voltage and parameter gain settings utilizes Flow Set fluorospheres If you wish to use a negative control sample use the appropriate AS Neg Ctrl Pro protocol to replace the Flow Set protocols in the above panels These negative control protocols have been provided Negative Control Protocols AS
50. at this screen if you adjusted the compensation of a Listmode file using the LMD QuickCOMP tool The original stored file remains unchanged and contains the Amplifier Gain information P1G FS Lin FCS Keywords Used in MXP Software See FCS Header Keyword Reference B 40 PN 177579BD USING MXP SOFTWARE FILE MENU Edit FCS Header Attributes Use this option to edit the FCS header information including calibration information You can change the displayed name of Parameters or change the scaling options Axis calibration PN 177579BD information can be entered here This information is embedded in the listmode file See Default Log Scaling for setting the default log tick marks W 1 File Edit FCS Attributes to view the Parameters on Edit FCS Header Attributes dialog box fe Name Fs Modes Linear x 2 Make the desired changes r Scale Use Default Scale Use Custom Scale Channel Value Units b foo Cancel Help 3 C or press to accept 4 D or press to exit the dialog box without saving the changes These changes may also be saved permanently within the open protocol by O File Save Protocol or D File gt Save Protocol As Note Modifying the scale options should not be done during acquisition Replaying of the listmode files always uses the calibration set at acquisition B 41 B USING MXP SOFTWARE FILE MENU Import Settings From C
51. axis tick marks B 99 dot plot title on output B 99 dot plot x and y axis labels B 99 FCS header legend plot B 118 legend plot phenotype logic columns B 118 overlay legend pane B 52 overlay statistics pane B 52 shutter for second laser 1 3 side scatter SS definition GLOSSARY 6 sensor definition GLOSSARY 6 sign file B 177 signal cables connections 2 3 signals processing of specifications 3 3 signatures See electronic signatures signing on to MXP software user ID B 3 PN 177579BD single file mode listmode playback B 91 Slider bars definition GLOSSARY 6 smooth density plot configuration B 109 histograms plot configuration B 102 plot display B 52 software options 1 3 specifications 3 5 warning statement xxiii See also MXP software solid state laser beam spot size 3 2 See also laser space needed instrument 2 1 special requirements installation 2 1 specifications instrument 3 1 Listmode 3 6 performance 3 11 physical 3 1 regions 3 5 software 3 5 See also performance specifications SQL definition GLOSSARY 6 stability check fluidics systems 1 5 check optical systems 1 5 day to day 3 12 performance specifications 3 12 within day 3 12 standard deviation SD See SD standard deviation start Cytometer B 171 startup wizard description B 2 red X appears B 2 Statistics print B 43 toolbar B 6 status bar AutoMATOR B 90 region editing B 137 PN 177579BD INDEX in
52. click on the file name to close the dialog box and display the file 3 The text box is placed in the upper left corner of the FlowPage a Use gt to move the text box to the desired location b D ouside the text box to anchor it See also Inserting a Line Inserting a Picture Inserting a Rectangle B 82 PN 177579BD USING MXP SOFTWARE B INSERT MENU Other Insert Menu Items Annotation This feature is not available in MXP software Auto Text This feature is not available in MXP software Overlay Histogram This feature is not available in MXP software FlowPAGE Analysis Results This feature is not available with MXP software FlowPAGE Analysis This feature is not available with MXP software PN 177579BD B 83 USING MXP SOFTWARE TOOLS MENU B 18 TOOLS MENU Publish Histogram Data Now This feature is not available in MXP software Publish Results Now Statistics are displayed under plots in resizable columns and can be exported to Microsoft Excel or ASCII text txt file format The Workspace Preferences Publish tab option Publish Data to Text File and or Publish Data to MS Excel must be enabled See Workspace Preferences Publish If publishing to Excel the default workbook file name consists of the Listmode file name with a time and date stamp added The file is saved to the user s Results directory The data can be published in Column Format or Experiment Table Format depending on your
53. colon or apostrophe Leading Trailing and multiple spaces are not allowed Open Histogram This feature is not available in MXP software B 30 PN 177579BD USING MXP SOFTWARE B FILE MENU Save All FCS Format Files This option saves the current listmode file to disk and saves any histograms marked for saving 1 Highlight the required plot xi P tagt 2 Data Source Histogram Scaling Note The individual Histogram s Plot ie sea A Far Properties must be set to save Acquisition Stop Condition IV Use Stop Condition Maximum Events 5000 IV Save Histogram Data in FCS Format Cancel Help Plots gt Acquisition Stop amp Save menu option N 3 D Save Histogram Data In FCS Format Note An S is added to the plot title if you select this option Do not confuse this with an S that identifies a gate 4 Repeat for each plot you wish to be saved 5 C File Save All FCS Format Files PN 177579BD B 31 USING MXP SOFTWARE FILE MENU Save Histogram File As The Save Histogram File As feature allows a single histogram associated with the currently in focus plot to be saved directly to an FCS histogram file Use the appropriate procedure depending upon whether you have the standard MXP software or also have the 21 CFR Part 11 option installed Select the appropriate link e Save Histogram File As Procedure With the 21 CFR Part 11 Option Installed e Save Histogram File As P
54. concerning the setting of this option can be found under Workspace Preferences Plot Display PN 177579BD B 167 USING MXP SOFTWARE CYTOMETER MENU B 168 Dots This sets the maximum events to be displayed during Setup Mode The default size is 1 000 events The Setup Mode rolling displays are updated up to three times per second from this buffer To change the size edit the Dots field to the number of Dots required If a zero is entered an error message displays showing range of 10 9999 The Dots value is stored in the Protocol Discriminators Lists the currently selected parameter and allows the setting and modification of discriminator values To change a value select the discriminator you wish to alter and use the Slide Control on the right hand part of the display to change the value as appropriate Parameter Button Selecting this displays the Setup Parameter Screen Red Laser Shutter Use to enable or disable the red laser The shutter setting is passed as part of the instrument settings part of the protocol Cytometer Control Parameter Setup This screen is used to select the Signals required in the Parameter Setup The signals that are available are selectable by checking the options you require Note If you add a parameter to an existing protocol the Detector names revert to the default names shown below IMPORTANT Risk of erroneous results if you assign the same parameter name to more than one detector Us
55. desired Gates Parameter and DataSource Histogram Scaling Regions for display File Gate Ungated hd Histogram Plot Data Source Histogram Formatting Paremete Fs Lin FS Lin Histogram Scaling Histogram Labeling JT Apply gate to all plats Stop and Save Regions Cancel Help B 100 3 When a Plot is displayed click the Copy Plot Image right mouse button to display further Copy Plot Data menu options Format Copy Plot Image Copy Plot Data Format Plot PN 177579BD USING MXP SOFTWARE B PLOTS MENU Histogram Plot Data Source Choose the parameter and gate for the current plot Gating equation and regions to be displayed can be selected File Histogram Plot Properties Gate Selection Parameter Apply Gate to All Plots Regions Parameter Select the required parameter for display from those available in the list Histogram Plot Configuration Use Gate Color Histogram Plot Properties Smooth Freeze Frame PN 177579BD B 101 USING MXP SOFTWARE PLOTS MENU B 102 Use Gate Color If the Use Gate Color option is checked the Histogram fill color is the same as the default color for the Gate that the plot is gated upon and the Color Select button is unavailable To change the color of the Histogram in the selected plot clear the Use Gate Color checkbox and click the Color Select button The colors returned are only those which the display is capable of ren
56. disappear See also Font PN 177579BD B 63 USING MXP SOFTWARE VIEW MENU B 16 VIEW MENU Acquisition Manager Note If no Worklist is visible D View gt Acquisition Manager to display the Worklist pane The Acquisition Manager is used to view and print a Worklist for the Cytometer The information to create a Worklist is defined in the Plate Wizard Once created the Worklist cannot be edited Print Worklist Click the right mouse button in any Acquisition Manager column heading J Berit workist to access the Print dialog box and print the Worklist See also ACQUISITION MANAGER TOOLBAR Worklist Columns Available Acquisition Manager Panel CREATING NEW PLATE WORKLIST CREATING PANELS Worklist Actions When you run a sample plate The well in process is highlighted and an arrow appears next to it e Completed wells remain highlighted B 64 PN 177579BD USING MXP SOFTWARE B VIEW MENU Worklist Columns Available The following columns appear within the Acquisition Manager Panel Protocol Region Settings Region Filename Instrument Settings Instrument Filename Parameter Names Well Loc Plate ID Sample IDs LMD File Name Cal Factor Reordering Columns The columns may be reordered by selecting a column or a group and using Drag and Drop to move it to the desired location To move a column within a group use Drag and Drop To move a group of columns use Ctr Drag and Drop Groups are de
57. gt gt Change Password Enter your current password in the Enter Old Password field Note If you reach the limit of password attempts allowed you are not able to log into the MXP software You must ask the System Administrator to unlock you and or reset your password before you can log into MXP software Enter your new password in the Enter New Password field Note Passwords must be a minimum of 6 characters and a maximum of 24 characters Blanks are not allowed For best security create a password with at least eight characters including letters numbers 0 9 and or symbols Re enter your new password in the Re enter New Password field Password Change Enter New Password Re enter New Password fil 5 Ome PN 177579BD USING MXP SOFTWARE B 21 CFR 11 MENU Sign File Version Use this feature to electronically sign an individual version of a listmode histogram protocol panel or PDF file e One or more persons can sign each version of each file e Each person can sign for the same reason or for a different reason e You can view the signature information for any version of a file by clicking on the Signed field in the Sign Versions dialog box or the appropriate Open File dialog box This feature is not available during any of these automated activities Acquisition Listmode playback Batch AutoMATOR and AutoSetup Procedure To sign a version of a listmode histogram protocol panel
58. in the MXP software Audit Trails can display Login Logout file handling automated and administrative activities for specified dates and users See the System Administrator s Guide for additional details Audit Trails are also provided by the DaCS software for MS Excel files See Audit Trails DaCS Software Note If you use any third party software application such as Adobe Acrobat you need to have procedures that ensure the security of these electronic records and provide an audit trail Security software applications may be purchased to provide security when using third party software applications ELECTRONIC SIGNATURES MXP software provides the capability to electronically sign individual versions of a listmode histogram protocol panel or PDF file See Sign File Version Excel worksheets can be signed electronically with the DaCS software provided with the MXP software Follow your laboratory procedures for signature approval for compliance See Electronic Signatures DaCS Software Note If you use any third party software application such as Adobe Acrobat you need to have procedures that ensure the security of these electronic records and provide an audit trail Security software applications may be purchased to provide security when using third party software applications DaCS SOFTWARE DaCS software is provided with the 21 CFR Part 11 option of the MXP software By providing Audit Trails and electronic signatures fo
59. mode allows for single file or panel based listmode file playback similar to acquisition of samples A single panel may be selected multiple copies of this panel may be run at a time Samples may be added or removed from individual panels Load Queue loads a listmode queue file a list of listmode files Replay Plate E loads a WLS file containing all the listmode files for a sample plate Regions may be Loaded or carried within a panel AutoGating and positives analysis can be active on any protocol within a panel Where the source listmode file is a 20 bit data file comp settings can be Run Time as stored within the LMD file or the Compensation settings file created in ListComp Setup or any CMP file may be used During playback the title bar displays the current file being analyzed Optionally a new file may be saved containing newly compensated 10 bit data The original LMD file cannot be overwritten automatically Saved files have the suffix entered in the Options tab appended to each saved file PN 177579BD B 91 USING MXP SOFTWARE TOOLS MENU B 92 List Comp Setup ei U ae ne S Single File Mode List Comp Setup Options Panel Filename Compensation Filename I Auto Save Trem Pretec ron 10 Fear This dialog provides an Acquisition Manager style worklist for the selection of compensation panels and associated protocols ppp files created from the protocols used in the AutoSetup applications d
60. mouse on the down arrow symbol a list of previous actions that can be Redone is displayed B 6 PLOT OPTIONS TOOLBAR PN 177579BD m Color Dot Plot Create a Color Dot Plot and specify the plot properties Histogram Plot Create a Histogram Plot and specify the plot properties Contour Plot Density Plot i A Overlay Plot Not available in MXP software Create a Density Plot and specify the plot properties Not available in MXP software Tomogram Plot Not available in MXP software Surface Plot xa Prism Plot Legend Plot Not available in MXP software Create a Prism Plot and specify the plot properties Create a Legend Plot and specify the plot properties B 5 B USING MXP SOFTWARE REGIONS OPTIONS TOOLBAR B 7 B 8 B 6 REGIONS OPTIONS TOOLBAR ts ia E E e ity che afe a Polygonal Region Rectangular Plot Quadrant Region Elliptical AutoGate Contour AutoGate Edit Both Prism Dividers Single Linear Region Multiple Linear Region Edit Single Prism Divider Insert a Polygonal Region into a dual parameter plot The region is a free form with up to 100 points It can include horizontal acute angles but not vertical re entrant angles Insert a Rectangular Region into a dual parameter plot Insert a Quadrant Region into a dual parameter plot All quadrants comprise four sets of Rectangular Regions on a dual parameter plot Create an Elliptical AutoG
61. operating at the time determines whether a particular Toolbar display is available Toolbars can be moved using the Drag and Drop method with the mouse around the MXP software desktop or customized to suit the preferences of each user See Toolbars Customize Toolbars The Toolbar elements are FILE OPTIONS TOOLBAR PLOT OPTIONS TOOLBAR REGIONS OPTIONS TOOLBAR GATE COLOR STATS AND HELP TOOLBAR FLOWPAGE TOOLBAR ACQUISITION MANAGER TOOLBAR AUTOMATOR TOOLBAR CYTOMETER TOOLBAR LISTMODE PLAYBACK TOOLBAR PN 177579BD USING MXP SOFTWARE FILE OPTIONS TOOLBAR B 5 FILE OPTIONS TOOLBAR The following Icon Buttons are shown within the File Options Toolbar Open T Save iF shee a Print 2 Cut Copy e Paste Sje Undo ou Redo Open the Open Listmode Data File dialog box Save all the FCS listmode data files that are currently open Save the Protocol with the current name If the Protocol has not yet been Saved the Save Protocol As dialog box is displayed Open the Print Plot printing dialog box Cut an item previously selected and place it on the Clipboard Copy a previously selected item into Clipboard while retaining it in its original position Paste a previously Cut or Copied item from Clipboard into the desired position Undo the previous command By clicking the mouse on the down arrow symbol a list of previous actions that can be Undone is displayed Redo the previous Undo command By clicking the
62. or PDF file 1 O 21 CFR 11 gt Sign File Version 2 The Sign Version dialog box appears Navigate to the directory needed Looki sa PanelABC tdf 20051002_160123 ix fe 01_5c 21 as flow set imd l 06_5c 21 as pe comp imd be 02_5c 21 as fite comp Imd l 07_5c 2l as verify 8 19 3 45 4 Imd le 03_5c 21 as pe comp Imd be 04_5c 21 as ecd comp Imd be 05_5c 21 as ape comp Id Type of signature X Sign Close PN 177579BD B 177 USING MXP SOFTWARE 21 CFR 11 MENU B 178 Select the file to be signed All of that file s versions appear Select the version to be signed Select a Type of signature from the pull down menu Note The use and significance of these codes is to be established by your laboratory The choices are Completed Verified Validated Authorized Approved and Certified D Sign Look in G PanelABC tdf_20051002_160123 gE le 01_5c 21 as flow set md ke 06_5c 21 as pe comp Imd le 02_5c 21 as fite comp Imd ke 07_5c 21 as verify 8 19 3 45 4 Imd lk 03_5c 21 as pe comp lmd lle 04_5c 21 as ecd comp lmd ll 05_5c 21 as ape comp Imd Version User 13 Oct 2004 12 13 10 5 Gladstone added stats displayed 4 Gladstone 13 Oct 2004 12 12 55 printed PDFs 3 ladstone 13 Oct 2004 12 12 47 exported data 2 Gladstone 13 Oct 2004 12 12 32 File imported into BCAP 1 Gladstone 13 Oct 2004 12 12 32 File imported into BCAP Type of signature
63. playback B 90 menu B 84 menu AutoMATOR B 85 publish histogram data now B 84 publish results B 84 support for 21 CFR Part 11 C 1 total events status bar B 174 tube sample See test tube tubing Power Supply pressure connection 2 5 Power Supply vacuum connection 2 5 waste container connection 2 5 INDEX 22 U uncontrolled file definition B 71 how to check in B 71 undo edit menu B 59 universal power supply dimensions 3 1 See also Power Supply unpacking of instrument 2 1 user ID admin B 2 administrator B 2 at software installation B 2 LMD file name B 46 signing on to MXP software B 3 user info cell type B 51 experiment B 51 institute B 50 institute address B 50 project B 51 sample source B 50 workspace preferences B 50 using MXP software B 1 V ventilation requirements 2 1 verification protocols 3 7 3 8 Vernier gain specifications 3 3 version display all B 24 display signed ones B 25 file sign B 177 file view B 177 version number definition GLOSSARY 6 view FCS header information B 40 menu B 64 menu customize toolbars command tab B 73 menu customize toolbars tab B 72 menu refresh B 73 menu resource explorer B 70 menu zoom B 73 view modify regions B 138 delete region B 141 PN 177579BD move region B 139 region name B 140 region points B 139 resize B 139 select region B 138 voltage pulse signals definition GLOSSARY 6 volt
64. protocol save and restore queries or errors can easily be identified and perhaps even explained or corrected by fax or telephone support Standard Default Files and File Extensions ALQ Queue of listmode files saved in AutoMATOR LMD FCS listmode data file includes ASCII header information and binary listmode data HST FCS histogram file single parameter only per file Includes ASCII text file descriptions and binary histogram channel information PGE FlowPAGE files PRO Protocol files PNL Panel files WLS Worklist files CMP Compensation files PPP Listmode compensation panel PDF Files created when Print to PDF is selected TDF Plate definition files Also applies to a folder that holds the listmode files for a plate KIT Reagent kit assay definition files WLQ Playback worklist files listmode files and protocols y p Output Txt files are created for the Plate Data Displays The following characters cannot be included in a file name forward slash backslash greater than sign gt less than sign lt asterisk comma question mark quotation mark pipe symbol colon or apostrophe Leading trailing and multiple spaces are not allowed PN 177579BD SPECIFICATIONS SOFTWARE SPECIFICATIONS Default AutoSetup Protocols and Panels Administrator The following AutoSetup panels and protocols are included with MXP software These files
65. rm Blend Gate 3 Gate Names New Gate 3 z Blend Color M Enable r Selected Population Name Color Group 7 ma M Logic C1 AND C2 AND C3 Create Gate Cancel Help 4 D or press to accept the changes D or press to exit the dialog box without saving the changes See also Create Modify Gates Workspace Preferences Gating PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Gate Names Allows the selection of a Gate that defines a population of interest Enable Enables the selection of one two or all three blend Gates Blend Color Displays the color to be associated with the selected Gate logic Name Allows a user defined name for the selected combination Color Allows you to modify the predefined colors Logic Displays the full coloring Gate logic for the selected population Create Gate Creates a logical Gate based on the coloring logic A Create Gate screen is displayed showing the Gate Logic to be applied Press Yes to Create the Gate or No to cancel PN 177579BD B 129 USING MXP SOFTWARE ANALYSIS MENU Modify Color Precedence When a subpopulation is very small and a larger blended color would swamp a smaller population Color Precedence can be used instead of Color Blending In Color Precedence Mode the subpopulation or color given the highest precedence overrules any color with lower precedence when the same event satisfies more than one color gate or
66. run and then pause before printing This allows you to make any necessary alterations to the plots and regions before outputting results Report Icon Once the files have been panelized the report icon only appears next to the last file as a report print out would only be produced after all files in a group have been run This differs from Acquisition Manager where there is a printout after each test that has run This is providing that the correct Output Options have been selected Once the files have been panelized you can select all or a block of the files so that they are highlighted If the icon on one of the highlighted files is changed all of the other selected files change to the same icon simultaneously To change the status settings for one file only highlight the file and then click on the icon until it changes to the desired status Reordering Files Once all files are listed they can be arranged in File Name or Date and Time order by clicking on the relevant column title Rearrange files by Dragging and Dropping them 1 on the relevant file so that it is highlighted 2 Hold the mouse button down and drag the file to the appropriate position so that it is between two file names 3 Release the mouse button If there are a number of files in the AutoMATOR Setup dialog box you can either use the scroll bar or utilize the Drag and Drop option B 87 B USING MXP SOFTWARE TOOLS MENU B 88 Select
67. selection in the Workspace Preference Publish tab To open the Statistics Pane move the mouse over the Plot so that the gt Sigma Cursor is displayed and click the left mouse button The Statistic Pane drops down showing ALL the Plot statistics To hide the Statistics Pane click the mouse on the Sigma cursor again Ungated 55 Lin FS Lt If other Regions are added or deleted while the Statistics Pane is open it is automatically resized to accommodate the change Plots including the Statistics Pane can be resized at Region Gated ALL 100 00 465 4 633 0 any time by dragging any part of the Window Frame K 100 00 4654 6330 although the height of the Statistics Pane remains Lymphs 25 33 130 6 449 7 unchanged The Statistics Pane can only be resized by using the 3 Splitter Bar while holding down ct Use the mouse to drag the revealed Statistics Pane to the required position If resizing is achieved using the splitter bar the automatic sizing using the Sigma Cursor is overridden Auto sizing is the default mode selected when the Plot is first created whereas user defined sizing is selected as soon as resizing occurs using the Splitter Bar To reinstate the Auto sizing hold down while clicking the gt Sigma Cursor with the mouse B 84 PN 177579BD USING MXP SOFTWARE B TOOLS MENU AutoMATOR Welcome to AutoMATOR Use the AutoMATOR option to sequentially read analyze and report a set of listmode fi
68. serial number e Software version Note If you have the 21 CFR Part 11 option installed on your system all printed FlowPAGEs also display the version number of the Listmode and Protocol files PN 177579BD USING MXP SOFTWARE INSERT MENU FlowPAGE from File When a FlowPAGE has been created including all the departmental logos addresses plots statistics and comment boxes the whole FlowPAGE can be saved and retrieved at will Use the open FlowPAGE option to open a FlowPAGE template from disk FlowPAGE uses the default file extension PGE The PGE extension is only a suggested extension any standard DOS characters can be used for the full file name C Sharlene Open Flowpage File 2 x i in v z e be 1 Insert gt Open FlowPAGE From File to Bee GB Sheene ck AcquisitionProtocol C Panel view the Open FlowPAGE dialog box AnalysisProtocol C PDF HST E Results HTML E worklist Images E Headeronly PGE f s E z Files of type Flowpage Files pae x Cancel 2 C the desired drive in the Drive List pa box 3 the desired directory in the Folders List box 4 O the type of file in the Files List box and click 5 D the name of the file to view O Co Or Double click on the file name to close the dialog boxes and display the file PN 177579BD B 75 B B 76 USING MXP SOFTWARE INSERT MENU FlowPAGE Statistics Table This option is used to insert statistics into FlowPA
69. the Aquisition Manager to carry settings between protocols within a panel you should ONLY carry settings between protocols where the parameter number and order are identical a Retrieves the instrument cytosettings from an external file For example from an AutoSetup protocol another protocol or listmode file When the Use instrument settings from external file option is selected the external file name is displayed in the Instrument Filename field PN 177579BD Instrument Settings AS 2C Settings PRO AS 2C Settings PRO B 67 USING MXP SOFTWARE VIEW MENU Instrument Filename Displays the external file name containing the instrument cytosettings used for this sample See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST Acquisition Manager Well Loc Plate ID and Sample Info B 68 Well Loc Plate ID Sample ID1 Sample ID2 Sample ID 3 Sample ID 4 Calibrator cytokine tdf Well Loc Displays the row and column location of a well in a sample plate or a tube in the tube holder as programmed in the Plate Wizard for example Al E40 Plate ID Displays the plate definition file name tdf for the current sample plate saved in the Plate Wizard Sample IDs Up to four Sample ID fields are displayed You can edit Sample IDs 2 3 and 4 before data acquisition starts From 3 to 24 characters ca
70. the Auto Pause placing the Automator into Continuous Pause To restart click the Pause button again and the test resumes from the point at which it was paused PN 177579BD B 12 CYTOMETER TOOLBAR 4 Restart Acquisition Start Acquisition Stop Acquisition Abort Acquisition EE E Idle Mode m N G Prime Cleanse Cytometer Controls aa amp PN 177579BD USING MXP SOFTWARE CYTOMETER TOOLBAR Reset the current acquired events to zero and clear the current data in memory Acquisition restarts at zero events and continues until a stop condition is reached Start acquisition Stop the Acquisition of the current sample and output results Stop the Acquisition without saving data or outputting reports Can use with Reload Plate Place the Cytometer in Idle mode in order to perform various cleaning and replacing procedures Flush the sample line and flow cell with sheath fluid to declog or remove bubbles Flush the sample line and flow cell with cleaning agent Display the Cytometer Control dialog box to adjust the Cytometer settings Click the button again to hide the dialog box B 9 B USING MXP SOFTWARE LISTMODE PLAYBACK TOOLBAR B 13 LISTMODE PLAYBACK TOOLBAR The listmode playback tool utilizes two sets of buttons one for saving and restoring worklists panels populated with listmode files and one for saving and restoring queues lists of listmode files D l
71. the Product in which it is contained provided that it is used on the computer hardware and in the operating system environment for which it was designed Should the media on which your software arrives prove defective Beckman Coulter will replace said media free of charge within 90 days of delivery of the Product This is your sole remedy for any breech of warranty for this software Except as specifically noted above Beckman Coulter makes no warranty or representation either expressed or implied with respect to this software or its documentation including quality performance merchantability or fitness for a particular purpose No Liability for Consequential Damages In no event shall Beckman Coulter or its suppliers be liable for any damages whatsoever including without limitation damages for loss of profits business interruption loss of information or other pecuniary loss arising out of the use of or inability to use the Beckman Coulter Product software Because some states do not allow the exclusion or limitation of liability for consequential damages the above limitation might not apply to you General This agreement constitutes the entire agreement between you and Beckman Coulter and supersedes any prior agreement concerning this Product software It shall not be modified except by written agreement dated subsequent to the date of this agreement signed by an authorized Beckman Coulter Representative Beckman Coulter is not bound b
72. to Print Plots Choose ALL or CURRENT Plots Print Statistics e Statistics Select this option if you wish to Print Statistics e Aggregate Files This option groups together statistics from all the regions on a single file Otherwise statistics are printed grouped by plot Print FlowPAGEs Select this option to print FlowPAGEs Choose ALL CURRENT or a SELECTION To print individual pages you can enable the page number checkbox PN 177579BD B 43 B USING MXP SOFTWARE FILE MENU Print to PDF Print plots statistics or FlowPAGES in PDF format xi Print Plots F Pa T C Current m Print Statistics IV Statistics IV Aggregate Files Print FlowPAGES Note During installation an ACSPDF printer driver is added to the Windows Printers folder This driver is used for the Print To PDF options only If you require additional functionality to create PDF reports you need to use a full version of Adobe Acrobat software This additional software is not included with MXP Software If you install Adobe Acrobat 4 0 or higher you should use the Adobe distiller to create PDF files rather than the ACSPDF PDF compatible print driver If you continue to use the Print to PDF options within MXP Software when Adobe Acrobat 4 0 or higher is installed some Flowpages are cropped This issue does not occur when Acrobat Reader only is installed Workspace Preferences Introduction Use Workspace Preferences option to set t
73. use electronic records to meet its record keeping requirements Implementation and compliance of the 21 CFR Part 11 regulation is the responsibility of the laboratory creating and signing the electronic records Proper laboratory procedures and key features of the MXP software with the 21 CFR Part 11 option installed as described in this section can be used to achieve compliance with this regulation The FC 500 MPL with MXP software is a closed system per the definition stated in Section 11 3 Subpart A of the regulation Closed System environment in which system access is controlled by persons responsible for the content of electronic records on the system All FC 500 MPL users in your laboratory must be trained to properly and consistently use your laboratory s procedures regarding electronic records and signatures User training can be documented in the MXP software by the System Administrator Note If you have the 21 CFR Part 11 option do not use RXP or CXP Analysis only software on your Workstation RXP and CXP Analysis only software are not 21 CFR Part 11 compliant Any analysis done in those applications will not be tracked in the 21 CFR Part 11 option for MXP software C 2 INTEGRITY OF ELECTRONIC DATA The division of Beckman Coulter that produces the FC 500 MPL and MXP software uses quality controlled validated development and manufacturing processes Beckman Coulter Inc is certified as IS0 9001 1994 and ISO 13485 complian
74. value is calculated Log Lin Selecting the Log Lin mode calculates the geometric mean values by determining the arithmetic mean of the raw ADC channels and then converting this to its relative linear channel equivalent Automatic Statistic Percentiles This is the channel which divides the events in a region by the percentile value The default percentile is 50 the median but can be set to request any desired value Different values may be set for both the X and Y axis PN 177579BD B 151 USING MXP SOFTWARE ANALYSIS MENU B 152 Absolute Count Calibration Use absolute count calibration particles for the calculation of absolute numbers of cells present in a sample IMPORTANT Ensure at least 1 000 Flow Count fluorospheres are counted or erroneous results may occur As asample is analyzed the numbers of a particular cell type in a region and the numbers of absolute count calibration particles present in a CAL region are compared and the absolute concentration of the cell type is then calculated automatically Note e A CAL region MUST be set for the system to be able to calculate an absolute count See Setting a CAL Region e Listmode replay uses the runtime protocol CAL Factor unless you select a different CAL Factor before replaying the file e Ifthe CAL Factor is set to 0 or there is no CAL region present in a protocol that has Cells uL selected Error appears in the Cells pL field To display the absolu
75. 001061 2003 06 12 1624 624 LMD 04_Cytokine p 625 Calibrator R_1 D_ 8 vandy 00001062 2003 06 12 1625 625 LMD 05_Cytokine p 312 5 Calibrator R_1 D_ 16 vandy 00001063 2003 06 12 1627 626 LMD 06_Cytokine p 156 Calibrator R_1 D_ 32 vandv 00001064 2003 06 12 1629 627 LMD 07_Cytokine p 080 Calibrator R_1 D_ 64 vandy 00001065 2003 06 12 1631 628 LMD 08_Cytokine p 040 Calibrator R_1 D_ 128 vandy 00001066 2003 06 12 1633 629 LMD 09_Cytokine p 020 Calibrator R_1 D_ 256 vandy 00001067 2003 06 12 1634 630 LMD 10_Cytokine p sample 1 Specimen R_1 Undefined vandy 00001068 2003 06 12 1636 631 LMD 11_Cytokine p sample 2 Specimen R_1 Undefined vandv 00001069 2003 06 12 1638 632 LMD a the desired directory in the Save in list box b W the type of file in the Save as type list box Filename Calibrator A1 D_ 1 vandv 00001059 2003 06 12 1620 622 LMD c Inthe File name text box type the Save as ype IMD Fiestind a Cae desired File Name 3 W to save the file MXP software creates a file with the name that you specified and stores it in the drive and directory you specified in FCS format File Name A listmode file name can have up to 200 alphanumeric characters including the filepath However the following characters cannot be included forward slash backslash greater than sign gt less than sign lt asterisk comma question mark quotation mark pipe symbol
76. 1 Workspace Preferences Plot Display B 51 Show in Region B 52 Black Plot Backgrounds B 52 Opaque Region Labels B 52 Print Plot Frames B 52 Print Dots Black B 52 Show Overlay Legend Pane B 52 Show Overlay Statistics Pane B 52 Log Scale Density Plots B 52 Smooth Histograms B 52 Default Overlay Plot Background Color B 53 Default of Events on Dot Plot B 53 Baseline Offset B 53 Default Overlay Mode B 53 Default 3D Plot Mode B 53 Default Plot Size B 53 Workspace Preferences Gating B 54 Automatic Gate Creation B 54 Automatic Gate Maintenance B 54 Automatic Color Precedence B 54 Dot Plot Coloring Mode B 55 AutoGating Sensitivity B 55 Elliptical Sensitivity B 55 Contour Sensitivity B 55 Contour Travel B 55 AutoGating Failure B 55 Pause on Error B 55 Abort Plate On Error B 55 Ignore Errors B 55 CONTENTS CONTENTS B 16 Workspace Preferences Publish B 56 File References in Data B 56 Publish Data to Text File B 56 Publish Data to MS Excel B 56 Switch to MS Excel on Publish B 57 Publish to MS Excel Format B 57 Export Bitmap Size B 57 Log In B 57 Log In Procedure B 58 Log Out B 58 Exit B 58 EDIT MENU B 59 Undo B 59 Redo B 59 Cut B 59 Copy B 59 Paste B 60 Paste Special B 61 Delete B 61 Edit Font B 62 Edit Font B 62 Edit Font FlowPAGE B 63 VIEW MENU B 64 Acquisition Manager B 64 Print Worklist B 64 Worklist Columns Available
77. 10 1142 464 LMD 20_caspace pl Well 20 Specimen R_1 Undefined vandv 00000878 2003 06 10 1144 465 LMD A Filename 20_caspace pl Well 20 Specimen R_1 Undefined vandv 00000878 Open Files of type LMD Files Ind fes z _ cma Open Runtime Protocol b the type of file in the Files of type list box c W the name of the listmode file you want to view 3 W Open Runtime Protocol to replay the LMD into the plots in the runtime protocol If the open runtime protocol is not selected the LMD replays into the plots currently displayed on the workspace Note Listmode replay uses the runtime protocol Cal Factor unless you select a different Cal Factor on the Absolute Count Calibration dialog box PN 177579BD B 27 USING MXP SOFTWARE FILE MENU When loading listmode files which have a parameter order different from the current protocol you are warned with Parameter Mismatch e Ifyou select Continue the file is loaded and replaces the parameters with those of the listmode file e Ifyou select Abort the listmode file is not loaded Note Listmode Compensation must be performed using the runtime protocol or a protocol with equivalent parameters as the runtime protocol When loading listmode files from a different instrument This is not an FC 500 file appears Select Continue to load the file Save Listmode File As B 28 Use this Save As feature if at any time you decide to save your listmode
78. 11 option The session of Microsoft Excel you will work in will be under the control of the DaCS software e Use the MPLAnalysis Add In if you do NOT have the 21 CFR Part 11 option Both the MPL_DaCSAnalysis Add In and the MPLAnalysis Add In perform these functions e Produce specialized data displays in Microsoft Excel from MPL data acquisition e Organize the data from a sample plate running a single protocol for display in a spreadsheet e Search for the calibration data and uses it to generate a calibration curve It tries to fit the data with a third order polynomial fit The coefficients of fit are saved to calculate the concentrations e Organize the specimen and control data and generates the well plots and if applicable calculates the concentration for unknown specimens e Display a threshold value for wellplots default is the average of the wellplots and displays a red line at the threshold value in the wellplot and shades all values in the data columns that are above the threshold in red The threshold value can be modified PN 177579BD B 15 USING MXP SOFTWARE FILE MENU B 16 Working With MPL_DaCSAnalysis and MPLAnalysis Add Ins Perform this setup procedure in order to publish graphs and data spreadsheets into Excel from MXP software using the MPL_DaCSAnalysis or MPLAnalysis Add In You can use either Analysis Add In when publishing from any of these features conditions AutoMATOR LMD Playback Tool LMD file
79. 146 PRISM plot dividers B 114 quadrant region re naming B 141 redo B 59 regions B 138 delete region B 141 move region B 139 region name B 140 region points B 139 resize B 139 select region B 138 status bar B 137 undo B 59 elapsed time status bar B 174 PN 177579BD electrical input dedicated lines 2 2 non dedicated lines 2 2 special requirements 2 2 electronic signatures 21 CFR Part 11 C 4 for Excel files C 6 provided by DaCS software C 6 sign file version B 177 enable red laser B 168 establishing fluorescence and or light scatter target ranges log A 1 establishing HV total gain ranges log A 1 establishing peak position and HPCV target ranges log A 1 events color dot plot B 97 definition GLOSSARY 2 Excel 21 CFR Part 11 considerations C 3 Add Ins for data export B 15 audit trails C 5 Audit worksheet C 5 electronic signatures C 6 MPL Analysis Add In B 15 MPL DaCsS Analysis Add In B 15 Result worksheet C 5 software version 1 2 System worksheet C 5 Exit B 58 experiment user info B 51 export file bitmap size B 57 definition GLOSSARY 2 extension cord Caution 2 2 F FC 500 MPL components description 1 1 installation 2 1 manuals hard copy xxi monitor linearity 1 5 specifications 3 1 view of instrument 1 1 FCS definition GLOSSARY 3 extension for histogram file 3 6 PN 177579BD INDEX extension for Listmode data file 3 6 file format 3 5 format files s
80. 2 Color Cell Activation PRO E Common Acquisition Sc 21 as verify 8 19 3 45 4 demo PRO E Common Analysis 2C Cell Activation pro 4c 1l pro Cytokine pro carryover pro caspace pro setup def qc 2I flow check pro qc 1l flow check pro hene alignment pro cleanse water pro cleanse bleach pro as FIS comp pro as fl4 comp pro 2003 05 2 2003 05 2 2003 05 2 2003 05 2 2003 05 2 2003 05 11 2003 05 0 2003 05 0 2003 05 0 2003 05 0 2003 05 0 2003 05 0 2003 05 0 B Cell Activation NEW tdf B 2 Color Cell Activation PRO Hh Plots fl BE Regions H Gates y LMD Files Ys 5c 2l as verify 8 19 3 45 4 Imd The view that you see depends upon your particular access rights User levels show only a particular user Listmode Histogram and Protocol directory depending on the tab selected If you are a member of a particular Workgroup all the user folders within that Workgroup are visible The Support tab provides a link to the Beckman Coulter and ACS web sites as well as links to any ancillary Help files B 70 PN 177579BD USING MXP SOFTWARE VIEW MENU 21 CFR Part 11 Option Uncontrolled and Controlled Files Definitions Uncontrolled and controlled Panel Protocol and Listmode files can appear in the Resource Explorer Controlled Files A Panel Protocol or Listmode file that has a version number and has been checked into the BCAGP database Changes to a controlled file are saved to a differ
81. 2C neg control pro AS 4C 2L neg control pro AS 3C 1L neg control pro AS 5C 1L neg control pro AS 4C 1L neg control pro AS 5C 2L neg control pro PN 177579BD 3 7 3 SPECIFICATIONS SOFTWARE SPECIFICATIONS All the supplied protocols may be copied and modified care should be taken to ensure all protocols have the same axis calibration to maintain correct functioning of the Compensation Wizard Default AutoSetup Protocols and Panels All Users The following AutoSetup panels and protocols are included with MXP software in each User s directory set up by the System Administrator If you have the 21 CFR Part 11 option installed on your system see the heading Resource Explorer for instructions how to check in these files for your use Panel Name Flow Set Protocol Compensation Protocols Verification Protocol Settings Protocol AS 5C 1L PNL AS 5C 1L Flow Set Pro AS FL1 Comp Pro AS FL2 Comp Pro AS FL3 Comp Pro AS FL4 Comp Pro AS FL5 Comp Pro AS 5C 1L Verify Pro AS 5C 1L Settings Pro AS 5C 2L PNL AS 5C 2L Flow Set Pro AS FL1 Comp Pro AS FL2 Comp Pro AS FL3 Comp Pro AS FL4 Comp Pro AS FL5 Comp Pro AS 5C 2L Verify Pro AS 5C 2L Settings Pro Default Generic Panels and Protocols All Users The following generic panels and protocols are included with MXP software in each Users directory set up by the System Administrator If you have the 21 CFR Pa
82. 79BD B 57 USING MXP SOFTWARE FILE MENU Log In Procedure To log into the MXP software 1 W File gt Log In 2 Enter your User ID and password x C OK Username Gladstone Note If the 21 CFR Part 11 option is installed the System Administrator can Ca PEE log into the MXP software at this dialog Eea box if the system was manually locked by a user Log Out Allows a particular user to Log Out of MXP software so that another person can Sign On If you have the 21 CFR Part 11 option installed e Italso closes any open Excel workbook and e Italso automatically logs you out of the DaCS software application Exit This option closes down the MXP software and returns you to your Windows Desktop If you have the 21 CFR Part 11 option installed e Italso closes any open Excel workbook and e Italso automatically logs you out of the DaCS software application B 58 PN 177579BD USING MXP SOFTWARE EDIT MENU B 15 EDIT MENU Undo Edit gt gt Undo reverses software operations such as Region drawing Region label editing and FlowPAGE editing Each New operation is appended to an Undo list The valid Undo operations are then performed in reverse order with each Undo request Note Undo does not undo such operations as Save and Acquisition of data The Edit Undo list is cleared after each change of plot focus The Edit Undo function is applicable to the Window Menu Regions FlowPAGE and fonts Redo Cut Edit
83. 83 FlowPAGE Analysis Results B 83 FlowPAGE Analysis B 83 TOOLS MENU B 84 Publish Histogram Data Now B 84 Publish Results Now B 84 AutoMATOR B 85 Welcome to AutoMATOR B 85 AutoMATOR Setup B 85 Reordering Files B 87 Load Plate B 88 Add File or Add Files B 88 Remove Files B 88 Add Blank B 88 Panelize B 88 Load Queue B 88 CONTENTS xi CONTENTS xii B 19 Save Queue B 88 Print B 89 Restore Analysis Files on Startup B 89 Show full pathname B 89 Output Options B 89 AutoMATOR Status Bar B 90 Listmode Playback B 90 Single File Mode B 91 List Comp Setup B 92 Options Tab B 93 AutoSetup Application Definition B 94 AutoSetup Scheduler B 94 PLOTS MENU B 94 Plots Introduction B 94 Duplicate Plot B 94 Color Dot Plot B 95 Copy Plot Image B 95 Copy Plot Data B 96 Format Plot B 96 Dot Plot Data Source B 96 File B 96 Browse Button B 96 X and Y Parameter B 96 Gate B 96 Apply Gate to All Plots B 96 Regions B 97 Dot Plot Events B 97 No of Events to Show B 98 Apply to All Plots B 98 Dot Plot Resolution B 98 Dot Plot Labeling B 98 Show Plot Title on Output B 99 Show Axis Tick Marks B 99 Show Axis Labels B 99 X and Y Axis Labels B 99 Font B 99 Histogram Plot B 100 Histogram Plot Data Source B 101 Parameter B 101 Histogram Plot Configuration B 101 Use Gate Color B 102 Smooth B 102 Freeze Frame B 102 Histogram Freeze Frame B 102 Create a Histogram Freeze Fra
84. AS 2C Settings PRO AS 2C Flow Set PRO AS 2C Neg Control PRO AS 2C Verify PRO 45 4C 1L Yerify PRO AS 4C 1L Flow Set PRO AS 4C 1L Settings PRO AS 4C 2L Settings PRO a AS 4C 2L Flow Set PRO The Open Protocol File dialog box appears Ls LL LL E AS FL1 Comp PRO AS FL2 Comp PRO EELELEELEELEE E File name fac 2L Flow Check PRO 2 the file you wish to open from the Files of ype Protocol Fies pro E Ee p La Folders list box 3 Oma Note Double clicking with the mouse on the file name closes the dialog box and opens the file Save Protocol PN 177579BD Save Protocol Feature With the 21 CFR Part 11 Option Installed If your system has the 21 CFR Part 11 option installed using the Save Protocol button performs differently depending upon whether or not acquisition is in process e If Acquisition is NOT in process the Save Protocol button works the same as the Save Protocol As Procedure With the 21 CFR Part 11 Option Installed e If Acquisition is in process the Save Protocol button automatically saves a new version of the protocol with a Save Comment that states this was a system generated save ctr S is the shortcut option as is the Save Protocol button Note While acquiring a Worklist avoid selecting Save Protocol when the system is transitioning to the next tube in the Worklist Note If you open protocols from other Users folders and you wan
85. B 3 administration user ID available at installation B 2 user ID description B 2 administrator user ID description B 2 advanced precedence B 131 air conditioning special requirements 2 2 ALQ file extension 3 6 ambient temperature instrument 2 2 amplification fine specifications 3 3 high voltages specifications 3 3 linear specifications 3 3 analysis menu B 122 protocol B 12 software options 1 3 Analysis Add In See MPL DaCS Analysis Add In and MPL Analysis Add In APC definition GLOSSARY 1 append PDF file name PDF file name extension LMD file name B 47 apply to all plots color dot plot events B 98 archived files display B 24 PN 177579BD INDEX system administrator can restore B 23 Argon laser beam spot size 3 2 description 3 2 See also laser argon laser power 3 2 arrange icons Windows B 181 ASCII definition GLOSSARY 1 assay values definition GLOSSARY 1 Audit Excel worksheet C 5 audit trails 21 CFR Part 11 C 4 for Excel files C 5 provided by DaCS software C 5 AutoGate definition GLOSSARY 1 autogating failure B 55 abort on error B 55 ignore errors B 55 pause on error B 55 region creating B 135 sensitivity B 55 contour sensitivity B 55 contour travel B 55 elliptical sensitivity B 55 automated activities change password function B 175 lock software function B 175 sign file version not available B 177 system generated file comments C 3 automatic color precedence
86. B 65 Reordering Columns B 65 Acquisition Manager Panel B 65 Panel B 65 Acquisition Manager Protocol B 65 Protocol B 65 Acquisition Manager Region Settings and Filename B 66 Region Settings B 66 Region Filename B 66 Acquisition Manager Parameter Names B 67 Parameter Names B 67 Acquisition Manager Instrument Settings and Filename B 67 Instrument Settings B 67 Instrument Filename B 68 Acquisition Manager Well Loc Plate ID and Sample Info B 68 Well Loc B 68 Plate ID B 68 Sample IDs B 68 Acquisition Manager Cal Factor B 69 Cal Factor B 69 Acquisition Manager LMD File Name B 69 LMD File Name B 69 PN 177579BD B 17 B 18 PN 177579BD Resource Explorer B 70 21 CFR Part 11 Option Uncontrolled and Controlled Files B 71 Toolbars Customize Toolbars B 72 Toolbars B 72 Toolbar Name B 72 Show ToolTips B 72 Cool Look B 72 Large Buttons B 72 New B 72 Reset B 72 Toolbars Customize Command B 73 Categories B 73 Buttons B 73 Description B 73 Zoom B 73 Refresh B 73 INSERT MENU B 74 Blank FlowPAGE B 74 FlowPAGE from File B 75 FlowPAGE Statistics Table B 76 FlowPAGE Textbox B 77 FlowPAGE Plot B 78 Insert Plot B 78 Inserting Drag amp Drop Plot B 78 Other FlowPAGE Object B 79 Inserting a Rectangle B 79 Inserting a Line B 80 Inserting a Picture B 81 Inserting a Text File B 82 Other Insert Menu Items B 83 Annotation B 83 Auto Text B 83 Overlay Histogram B
87. CS Format checkbox Note The actual histogram save action is only performed if the Save Histogram checkbox is checked in Workspace Preferences Acquisition Options See also Color Dot Plot Histogram Plot Density Plot Format Plot Acquisition Stop and Save PN 177579BD USING MXP SOFTWARE PLOTS MENU Use Stop Condition Check this checkbox if the stop counter is to be activated Maximum Events Set the maximum number of events you wish to be recorded Save Histogram Data in FCS Format Check this checkbox to save the Histogram Data in FCS Format Format Plot To display the Plot Properties dialog box click the right mouse button on the plot and g Format Plot Format options differ depending on the Plot type Select a formatting option from the table below to view detailed information about the format options available for the different types of plots Plot Type Plot Name Available Formatting Options Single Parameter Histogram Plot Data Source Histogram Scaling Labeling Font Stop amp Save Dual Parameter Dot Plot Data Source Events Resolution Labeling Font Stop amp Save Density Plot Data Source Density Resolution Labeling Font Stop amp Save Special Types Legend Plot Data Source Options FCS Font Prism Plot Data Source Scaling Font PN 177579BD B 121 B USING MXP SOFTWARE ANALYSIS MENU B 20 ANALYSIS MENU Create Modify Gates Use the Gating function to define populations of in
88. Check the appropriate options for the actions you wish to perform when acquiring data using the Acquisition Manager See also Workspace Preferences Publish When you select Publish and data is exported to MS Excel the workbook file name consists of the Plate ID and a date time stamp The file is saved to the user s Results directory The number of worksheets produced depends on the number of protocols used in the worklist and whether you selected Column Format or Experiment Table Format on the Workspace Preferences Publish page Note Save Protocol applies to any changes made to a protocol whether acquiring or not You cannot alter the Save LMD default as this is a mandatory setting If a protocol name begins with Cleanse the protocol does not create a listmode file when the protocol is run If you have the 21 CFR Part 11 Option installed on your system and Save Protocol is selected a new version is created and saved at the end of acquisition for any protocol that was run A comment is saved with that file version stating it was system generated Automatic versioning also applies to the protocols and Instrument Settings files in the AutoSetup panel Acquisition Manager e Use Spaces Between Panels Not available for MXP software e Continue AutoSetup on FlowSet HV Setup Failure Eliminates the manual approval step in AutoSetup Wizard when running Flow Set fluorosphers Default Log Scaling Sets the default logarithmic scaling tick mar
89. Cytomics FC 500 MPL With MXP Software Reference PN 177579BD October 2011 Beckman Coulter Inc 250 S Kraemer Blvd OULTER Brea CA 92821 WARNINGS AND PRECAUTIONS READ ALL PRODUCT MANUALS AND CONSULT WITH BECKMAN COULTER TRAINED PERSONNEL BEFORE ATTEMPTING TO OPERATE INSTRUMENT DO NOT ATTEMPT TO PERFORM ANY PROCEDURE BEFORE CAREFULLY READING ALL INSTRUCTIONS ALWAYS FOLLOW PRODUCT LABELING AND MANUFACTURER S RECOMMENDATIONS IF IN DOUBT AS TO HOW TO PROCEED IN ANY SITUATION CONTACT YOUR BECKMAN COULTER REPRESENTATIVE HAZARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS WARNINGS CAUTIONS and IMPORTANTS alert you as follows WARNING Can cause injury CAUTION Can cause damage to the instrument IMPORTANT Can cause misleading results BECKMAN COULTER INC URGES ITS CUSTOMERS TO COMPLY WITH ALL NATIONAL HEALTH AND SAFETY STANDARDS SUCH AS THE USE OF BARRIER PROTECTION THIS MAY INCLUDE BUT IT IS NOT LIMITED TO PROTECTIVE EYEWEAR GLOVES AND SUITABLE LABORATORY ATTIRE WHEN OPERATING OR MAINTAINING THIS OR ANY OTHER AUTOMATED LABORATORY ANALYZER WARNING Risk of operator injury if e All doors covers and panels are not closed and secured in place prior to and during instrument operation e The integrity of safety interlocks and sensors is compromised e Instrument alarms and error messages are not acknowledged and acted upon e You contact moving parts e You mishandle broken parts e Doors covers and
90. DaCS Analysis Add In and MPL Analysis Add In manual lock out 21 CFR Part 11 C 2 manual updates iii manuals for your instrument about your reference manual xxi conventions used xxii how to use xxi introduction to your manuals xxi using the manuals xxi marker creating linked B 137 creating non linked B 137 material safety data sheets MSDS how to order 1 5 MB definition GLOSSARY 4 mean definition GLOSSARY 4 menu 21 CFR Part 11 B 175 analysis B 122 Cytometer B 164 definition GLOSSARY 4 edit B 59 file B 11 FlowPAGE B 154 insert B 74 plots B 94 tools B 84 view B 64 window B 179 Microsoft Excel See Excel minimum count B 136 minimum event counter definition GLOSSARY 4 mode cleanse use of cleaning agent 1 4 modify advanced precedence B 131 color blend B 128 color precedence B 130 PRISM plot B 147 regions B 138 PN 177579BD modify color blend blend color B 128 gate name B 128 monitor specifications 3 4 monoclonal antibody QC material used to verify performance 1 5 QC material used to verify staining lysing and analysis 1 5 mouse definition GLOSSARY 4 MPL Multi Plate Loader definition GLOSSARY 4 description 1 2 illustration 1 1 MPL Analysis Add In used in Excel B 15 MPL DaCsS Analysis Add In used in Excel B 15 MSDS material safety data sheets how to order 1 5 Multi Plate Loader MPL See MPL Multi Plate Loader multiple linear regions creating B 137 interacti
91. Data Format Copies the current plot image into memory for pasting into other applications See Copy for more details B 95 B USING MXP SOFTWARE PLOTS MENU B 96 Copy Plot Data Copies the raw plot data into memory for pasting into other applications such as Microsoft Excel Format Plot Displays the Plot Properties dialog box for the current plot See Format Plot Dot Plot Data Source Choose the X and Y parameters for the current plot Gating equation and regions to be displayed can be selected ie x G ate Labeling Font Stop and Save X and Y Parameter Data Source Events Resolution Apply Gate to All Plots ee wr Regions y x Parameter Y Parameter FL3 Log FL3 Log x FL4 Log FL4 Log x I Apply gate to all plots Regions Cancel Help File Select the required file from the files already in memory displayed in the drop down box or click the browse button to explore your directories for the required file Browse Button Allows you to peruse other directory options and select files as appropriate X and Y Parameter Select the required parameter for display from those available in the list Gate Available Gates are listed Gates required for Advance Precedence should be selected at this point See Modify Color Precedence Apply Gate to All Plots To select this option enable the checkbox and apply the gate selected to all plots PN 177579BD USING MXP SOFTWARE B PLOTS MENU
92. Default Plot Size Allows you to select the size of the plot display B 53 B USING MXP SOFTWARE FILE MENU Workspace Preferences Gating Automatic Gate Creation xi Automatic Gate Maintenance LMD File Name Acquisition Options User Info Automatic Color Precedence Plot Display Gating Publish Dot Plot Coloring Mode r Dot Plot Coloring Mode AutoGating Sensitivity pe een IV Automatic Gate Creation Elliptical Sensitivity Color Precedence Mode Contour Sensitivity IV Automatic Gate Maintenance IV Automatic Color Precedence Contour Travel m AutoGating Sensitivity AutoGating Failure Elliptical 011 z Pause on Error Contour fT Travel 180 Abort Plate On Error a Ignore Errors r AutoGating Failure Ignore Errors Pause On Error Abort Plate On Error Cancel Undo Apply Help See also Workspace Preferences Introduction Workspace Preferences LMD File Name Workspace Preferences Acquisition Options Workspace Preferences User Info Workspace Preferences Plot Display Workspace Preferences Publish Automatic Gate Creation This option allows you to create a new gate automatically when a new Region is created If this option is not selected you must use Analysis Create Modify Gates to create gating logic Regions copied using CtlJ Drag and Drop into a plot assign the region as a gate Automatic Gate Maintenance Enabling this option ensures that when the gating o
93. Drag amp Drop Plot Insert currently displayed plots on the desktop into FlowPAGE 1 W inside the plot to be inserted into FlowPAGE and hold down the left mouse button The No Drop cursor is displayed 2 Drag the cursor onto the FlowPAGE The Drop cursor fy is now shown Release the mouse button The plot is now inserted into FlowPAGE Note If you Drag and Drop the plot the plot and the statistics are inserted If you ctr Drag and Drop the plot the statistics are not inserted with the plot B 78 PN 177579BD USING MXP SOFTWARE B INSERT MENU Other FlowPAGE Object Inserting a Rectangle 1 D Insert Other FlowPAGE Object and the Rectangle option A cross hair cursor is displayed 2 and hold down the left mouse button while you drag to size the rectangle 3 J outside the rectangle to anchor it See also Inserting a Line Inserting a Picture Inserting a Text File PN 177579BD B 79 INSERT MENU USING MXP SOFTWARE B 80 Inserting a Line Draw straight lines using FlowPAGE Insert Line 1 D Insert Other FlowPAGE Object and W the Line option A cross hair cursor is displayed 2 Drag the cursor to desired line size and release the mouse button The line is drawn and the cursor reverts back to the Chooser cursor N See also Inserting a Rectangle Inserting a Picture Inserting a Text File PN 177579BD USING MXP SOFTWARE INSERT MENU Inserting a
94. FSET key value 3 10 CAROUSEL key value 3 10 COMPENSATIONMODE key value 3 10 CYTOMETERID key value 3 10 Discriminator key value 3 10 FILEGUID key value 3 10 LOCATION key value 3 10 PANEL key value 3 11 PnADDRESS key value 3 10 PnC key value 3 10 PnDIVIDER key value 3 10 PnGAIN key value 3 9 PnQ key value 3 10 PnU key value 3 10 PnX key value 3 10 PnZ key value 3 10 RATIODENOMINATORMUX key value 3 11 RATIONUMERATORMUX key value 3 10 INDEX 1 INDEX REDLASER key value 3 10 RESAVEDFILE key value 3 10 SAMPLEID1 key value 3 10 SAMPLEID2 key value 3 10 SAMPLEID3 key value 3 10 SAMPLEID4 key value 3 10 TARGETLASERPOWER key value 3 10 TRAYANALYSIS key value 3 11 TRAYID key value 3 11 TRAYROWCOL key value 3 11 TUBENO key value 3 10 Y2KDATE key value 3 10 Numerics 21 CFR 11 menu B 175 21 CFR Part 11 audit trail C 4 change password B 175 DaCs software C 4 definition GLOSSARY 1 electronic signature procedure B 177 electronic signatures C 4 integrity of electronic data C 1 lock the software B 175 menu B 175 option compliance information C 1 overview C 1 password change B 175 passwords C 1 sign file version B 177 third part applications C 3 use Windows security features C 3 A abort abort the MPL B 172 INDEX 2 abort the panel B 172 abort the tube B 172 on error autogating failure B 55 re
95. GE C 1 Insert gt gt FlowPAGE Statistics Table F1 Ungated File001 LMD FL1 LOG FL2 LOG E F1 Ungated File001 LMD Legend l t Overlay Plot 1 From the list of plots currently open in MXP software D the one containing the statistics you wish to insert 3 W or press to exit the dialog box without accepting the changes The statistics for the selected plot are inserted into the FlowPAGE and can then be moved to the desired position Only one plot is permitted per statistics table PN 177579BD USING MXP SOFTWARE B INSERT MENU FlowPAGE Textbox This option is used to insert text into FlowPAGE 1 Insert gt gt FlowPAGE Textbox An insert text cursor A is displayed 2 Move the cursor to the place for the textbox and O 3 Type the required text and Edit Text x Om OK Cancel 4 W outside the text box to accept the changes 5 Click the right mouse button on the text box to edit properties enter text edit the font or change the alignment PN 177579BD B 77 USING MXP SOFTWARE INSERT MENU FlowPAGE Plot Insert Plot To insert plots currently displayed on the desktop into FlowPAGE Select Plot to Use x 1 D Insert gt gt FlowPAGE Plot and F1 Ungated F1 SS FS O F1 Ungated F1 FL1 LOG FL2 LOG a plot in the list 2 Loo The plot is placed in the upper left corner of the FlowPAGE Cancel Hep Inserting
96. ILE MENU Open Protocol Use this feature to open a protocol file Use the appropriate procedure depending upon whether you have the standard MXP software or also have the 21 CFR Part 11 option installed Select the appropriate link e Open Protocol Procedure With the 21 CFR Part 11 Option Installed e Open Protocol Procedure Standard MXP Software Installed Open Protocol Procedure With the 21 CFR Part 11 Option Installed Use this procedure if you have the 21 CFR Part 11 option installed on your system Ask your System Administrator to restore any archived files that you need to access before proceeding Only the System Administrator can archive files and restore archived files 1 O File Open Protocol 2 Select the desired protocol file zi Look in AcquisitionProtocol x a 3c 1l pro R AS 2C Settings PRO Boas 3c Illy ae 4c 1l pro BE AS 2C Flow Set PRO BRAS 4CI1L ae 4c Alpro BE AS 2C Verify PRO BP as 4C1L a 5c ILpro B2 as 3c 11 settings pro BPs 4C1L H 5c 2l pro HH as 3c 11 flow set pro H AS FLI C C Show Archived Files if you want to display archived files in addition to the current protocol H File name enpo o Open files appears next to each Files of type Protocol Files pro x Cancel archived file name I Show Archived Files J the desired directory in the Look in list box W the type of file in the Files of type list box J the name of the protocol file y
97. MENU 1 Select the items you wish to change See Select a Group of Items Within FlowPAGE 2 J z All items are now vertically spaced equally Save FlowPAGE As To save an existing FlowPAGE as a FlowPAGE template 1 FlowPAGE Save FlowPAGE As to view the Save FlowPAGE dialog box 2 Specify the desired drive and folder by selecting them in the list boxes or by typing the path in the File text box 3 In the File name text box type the desired File Name Save in Sy admin he Al el I Hst A acs L Html C Results J Images C Worklist Protocol Fiemme PS Save as type Flowpage Files pge bal __ Cancel FlowPAGE gives the default file extension PGE The PGE extension is only a suggestion any standard DOS characters can be used for the full file name Note Keep the extension consistent for easy file searches B 163 B USING MXP SOFTWARE CYTOMETER MENU B 22 CYTOMETER MENU The Cytometer menu is concerned with online connection and control of the flow cytometer The Cytometer Control menu option allows the Cytometer Control screens to be activated Until this is done the other Cytometer menu items are grayed out The Status Page displays current instrument settings which can be printed out for future reference The Cytometer Log displays a log of errors generated by the system The Get Cytosettings from Protocol option allows Cytometer Instrument Settings to be obta
98. MP File This option allows compensation settings obtained from the Listmode Playback option to be imported into the current protocol 21x 1 File Import gt Settings from CMP Look Gi Acton rotoos ce File AS 4C 1L Flow Set cMP i AS 4C 2L Flow Set cMP 2 AS 5C 1L Flow Set CMP 2 AS 5C 2L Flow Set CMP R C File name 2 the appropriate file and Files of type LMD Compensation Files cmp m __ Cancel ZA The compensation settings from the stored file are then read into the current MXP software protocol B 42 PN 177579BD USING MXP SOFTWARE FILE MENU Print You can print plots and statistics in a variety of ways m Printer 1 File gt Print The Print dialog box m Name Canon GP300 405 PCL x Properties is displayed Status Default printer Ready Type Canon GP300 405 PCL Where SBSERVER CanonGP3 Comment J Print to file Print Plots 7 Copies All 2 Make the desired changes vie Si Number of copies M H Curent I I i Print Statistics s LH alel ale I Collate M Statistics IV Aggregate Files m Print FlowPAGES M FlowPAGES Cancel C All v PAGE 1 PAGE 2 C Current inl 3 O or press Enter to V PAGE 3 TEON Te Selection commence printing See also Print Plots Print Statistics Print FlowPAGEs 4 D or press to exit the dialog box without saving the changes Print Plots Select this option if you wish
99. N 177579BD B 123 USING MXP SOFTWARE ANALYSIS MENU Gate Color When a region is drawn on a plot the first region is automatically shown as red meaning events inside this gate are colored red when viewed on a color dot plot The second is in green and the third in blue These colors can be altered through the Color function if more or different colors are required 1 w the down arrow next to the Color Well for the selected Gate or select the Ungated events color down arrow to change the color for Ungated events 2 D the required color This automatically changes the events within the gate to this color Ein 1s al 188 1m itm 1118 ther i 3 If further colors are required D Other to reveal a larger color palette C a defined color or drag the cursor around on the color board to select the color of choice Hue 0 Bed Sat 2 Green 0 Jo Ea a Blue Jo Add to Custom Colors B 124 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU 4 When the appropriate color has been found you can click the Add to Custom Colors button for easy use at a later date or you can E to add to a plot The same principles can also be applied to the Ungated Events Color option Boolean Gating MXP software implements Boolean gating Up to eight regions can be combined to form a gate A gate is used to select a specific subpopulation of data MXP software uses a combination of regions and operators to achi
100. NTS 2 2 Electrical Input CAUTION Possible instrument damage could occur if you put the Power Supply plugs on the same electrical circuit or use an extension cord or a power strip to connect the Power Supply Use two dedicated outlets with isolated grounds for the Power Supply plugs Country Dedicated Lines with Isolated Grounds Non Dedicated Lines USA One dedicated line at 115 Vac 50 60 Hz at One non dedicated line at 115 Vac 20A 50 60 Hz at 20 A Europe and Two dedicated lines at 220 Vac 50 60 Hz Three non dedicated lines at 220 Vac other applicable at 20 A 50 60 Hz at 20 A countries or Two dedicated lines at 240 Vac 50 60 Hz at 20 A Japan Two dedicated lines at 100 Vac 50 60 Hz Three non dedicated lines at 100 Vac at 20A 50 60 Hz at 20 A USA units are shipped with a line conditioner only requiring one line The computer requires a separate outlet but it does not have to be a dedicated line Ambient Temperature and Humidity Keep the room temperature between 16 C and 32 C 60 F and 90 F and do not let it change more than 5 F per hour Keep the humidity between 30 and 85 without condensation Heat Dissipation Heat dissipation is 2 300 W 7 850 Btu hour for a single laser system Heat dissipation is 2 330 W 7 955 Btu hour for a two laser system Provide sufficient air conditioning refer to Ambient Temperature and Humidity Drainage The waste line from the Cytometer i
101. No Drop Cursor is displayed 3 Drag the cursor onto the FlowPAGE The Drop Cursor is now shown 4 Release the mouse button to paste the Plot Image plus the Statistics onto a FlowPAGE Note If you use v to Drag and Drop you get the Plot Image but not the Statistics B 60 PN 177579BD USING MXP SOFTWARE B EDIT MENU e Ifthe Paste target is a Plot then regions are Pasted 1 Select the Plot with the region of interest to be copied 2 Drag the cursor to the destination plot 3 Release the mouse button to paste the Regions onto the Plot Note If the copied region is required as a gate use oO to Drag and Drop the region See also Cut Paste Paste Special Paste Special Allows which form of a copied object is pasted Bitmaps Text or Region may be pasted depending on the copy source and the paste target See also Cut Copy Delete When Delete is used any selected items are deleted In certain circumstances it may be possible to reverse the Delete action by selecting the Undo option See also Undo PN 177579BD B 61 USING MXP SOFTWARE EDIT MENU B 62 Edit Font Edit Font You can modify the Font type size and color of selected text The fonts available depend upon the fonts installed in Windows If in doubt try to use only True Type fonts The fonts selection dialog box is the standard Windows common dialog box for font selection and is fully explained in the Mic
102. Open Protocol Procedure Standard MXP Software Installed B 37 Save Protocol B 37 Save Protocol Feature With the 21 CFR Part 11 Option Installed B 37 Save Protocol Procedure Standard MXP Software Installed B 38 Save Protocol As B 38 Save Protocol As Procedure With the 21 CFR Part 11 Option Installed B 38 Save Protocol As Procedure Standard MXP Software Installed B 39 Save Listmode Compensation As B 40 View FCS Header Information B 40 FCS Keywords Used in MXP Software B 40 Edit FCS Header Attributes B 41 Import B 42 Settings From CMP File B 42 Print B 43 Print Plots B 43 Print Statistics B 43 Print FlowPAGEs B 43 Print to PDE B 44 Workspace Preferences Introduction B 44 Workspace Preferences LMD File Name B 45 First 10 characters B 46 Sample ID 1 2 3 4 B 46 Well Loc B 46 User ID B 46 Run Number B 46 Y2K Date B 46 Time B 46 PN 177579BD PN 177579BD Tag Number B 47 File Name Extension B 47 PDF File Name Extension B 47 Append PDF File Name B 47 Replace B 47 Next File Name B 47 Workspace Preferences Acquisition Options B 48 Output Options B 49 Acquisition Manager B 49 Default Log Scaling B 49 Cytometer Control Translucency B 49 Workspace Preferences User Info B 50 Institute B 50 Institute Street B 50 Institute Town B 50 Sample Source B 50 Project B 51 Cell Type B 51 Experiment B 51 Cytometer Serial Number B 51 Currently Logged On User B 5
103. Picture into a FlowPAGE Add a Text Box to a FlowPAGE Draw a Line on a FlowPAGE to enhance the graphical layout of the page Insert a Rectangle into a FlowPAGE that can be used as a border for Pictures or Text Boxes Align all the selected items to the top of the selection area Align all the selected items to the bottom of the selection area Align all the selected items to the left of the selection area Align all the selected items to the right of the selection area Space selected items within a FlowPAGE evenly across a page Space selected items within a FlowPAGE evenly down a page Resize the selected items to all be the same size as the reference item Resize the selected items to all be the same size as the reference item B 10 ACQUISITION MANAGER TOOLBAR PN 177579BD New Plate E Reload Plate Reanalyze Current Plate Print Open the Plate Wizard Copy the Worklist from the last plate that was run Not available in MXP software Print the current Worklist Note The Listmode File Name column displays only a provisional file name including the well number prior to acquisition If Time or Run Number is included as part of the name the actual value of these fields can only be determined once acquisition of each sample is complete Therefore if you wish to print a worklist including the final listmode file names only do so once acquisition of all samples in the Worklist is complete B 7 B USING MXP SOFT
104. Region is deleted any Gate or statistics that used the Region are also automatically deleted When a Gate has been deleted any plot that used the deleted gate is reset to Ungated MXP software allows you to resize an already created Region Rectangular Region This option draws a Rectangular Region on a dual parameter plot See also Create Rectangular Regions in the Instructions For Use manual Rectangular Region Properties Region Name x Prime Region Name K Automatic Region Name Positioning X and Y Coordinate J Prime IV Automatic region name positioning i j i iti Lower left Interactive Linear Region Editing X Coordinate 673 Y Coordinate eos Upper right X Coordinate jpo Y Coordinate jee Minimum Count IT Active Count f0 Cancel Help PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Interactive Rectangular Region Editing When a Region s position is changed any Gates and statistics that use the Region are automatically updated and each plot is redrawn to reflect the changes in the Gate When the Region is deleted any Gate or statistics that used the Region are also automatically deleted When a Gate has been deleted any plot which used the deleted Gate is reset to Ungated MXP software allows resizing of previously created Regions Quadrant Region Inserts a Quadrant Region into a dual parameter plot All quadrants comprise four sets of Rectangular Regions on a dual parameter plot The Quadrants are a
105. This refers to the Processed Data Rate that is the number of events which have exceeded the discriminator subsequently processed and saved in memory Flow Rate This item shows the relative flow rate of the Cytometer during acquisition Click on the drop down box and Ste choose the required rate Cytometer Window Help The Flow Rate can only be displayed ES E E as Low Medium or High The Flow j m Medium gt Rates are approximately 10 30 and i 60 pL min respectively aA Cytometer Status Displays the current status of the Cytometer and the most recent error message Double click on this field to display the Cytometer Status Message dialog box Windows Status Bar The Windows Status Bar normally gives a brief description of selected menu items When drawing or editing Regions the Status Bar shows Channel for X and Y parameters respectively as they are drawn B 174 PN 177579BD USING MXP SOFTWARE B 21 CFR 11 MENU X and Y Coordinates The X and Y channel values are the coordinates of the cursor position given by a scaling factor This scaling factor depends upon whether the respective parameters are linear or logarithmic amplification and whether you have calibrated the parameters or not If the parameters are linear and uncalibrated the X and Y channels vary between 0 255 or 0 1024 depending on the data source If the parameters are loga
106. Tile PN 177579BD B 179 B USING MXP SOFTWARE WINDOW MENU Tile Special Tile Special overrides the normal Windows Tile command this allows plots within the workspace to be arranged in a logical manor Plots are sized equally according to the default size setting in File Workspace Preferences gt gt Plot Display The plot size in Tile Special resizes the plots currently displayed on the workspace New Plots may be ordered by File Gate or Parameter or any combination of these At the start of each row the Plot Type Icon is displayed 1 D Window gt gt Tile Special to display the Tile Special dialog box 2 By clicking the mouse on the File Gate or Parameter column heading the plots are displayed across the screen in alphabetical order Clicking the appropriate column heading again displays the plots in reverse order 3 The order is shown in the Tile Special dialog box W to apply the change You can also Drag and Drop the plots into any order B 180 Tile Special x Fe o aae e r O1_Well 01 Specimen R_1 Undefined vandv 262 LMD Ungated L 01_Well 01 Specimen R_1 Undefined vandv 262 LMD CELLS FL FLOWPAGE2 1 Plot Size C Small 180x180 Medium 225x225 C Large 300300 Cancel Hep PN 177579BD USING MXP SOFTWARE WINDOW MENU B The d icon is activated by clicking to the left side of a plot listed in the File column This forces that plo
107. WARE AUTOMATOR TOOLBAR B 11 AUTOMATOR TOOLBAR The AutoMATOR Toolbar allows shortcuts to commonly used AutoMATOR functions bim 501_9C 1L Flow Test Spx AutoMATOR Setup Display the AutoMATOR Setup dialog box Once all alterations have been made D Oox The toolbar becomes active once all files have been entered A list of the first file for each panel usually the Control files is shown allowing you to choose the batch required to run first or_ac 1L Flow Test Sp The rest of the files follow in order after the selected panel B 8 Stop Run Pause Cancel the batch run This aborts the current panel When the panel runs again the test starts from the beginning Start the batch running The AutoMATOR Status Bar appears showing the status of the tests that is if it is currently paused still running and so on Pause has two states Continuous Pause Pause the tests for modifications and remain on pause until you decide that the test will continue This function allows the test to stop without reverting to the beginning of the panel again To resume the test click the Pause button again Auto Pause The run of tests will pause and then carry on with the rest of the panel automatically after a 15 second wait The countdown is displayed on the Automator Status Bar When the tests are in the Auto Pause setting you can hold down the key and click the Pause button The key overrides
108. a file using the mouse keep the mouse button held down and drag to either the top or the bottom of the window Moving the cursor outside the window scrolls the list The further the cursor is pushed to the top bottom the faster it scrolls Load Plate Displays a dialog box to select and load a plate definition file tdf or worklist file wls This loads all of the files generated by the corresponding worklist Add File or Add Files Select this option to display the Open Listmode File dialog box e 21 CFR Part 11 Option You can then search for other Files in various drives and directories Select the name of the listmode file you want to add You can only add one listmode file at a time The most recent version of the listmode file is opened You can repeat this procedure to add multiple files to the AutoMATOR Setup screen e Standard MXP Software You can then search for other Files in various drives and directories and add them to the list Select the name of the listmode file s you want to add Remove Files When selected this option removes any highlighted files within the AutoMATOR Setup dialog box The button performs the same function Add Blank A Blank is used to fill a space when there is an incomplete panel of listmode files so that the results from the tests are kept in the correct order and a complete panel is maintained When selected this option puts a blank at the end of the file list This can then be Dragged
109. accuracy PRIME region When a histogram Peak is not within a PRIME Region the system performs an AutoPrime Printer An optional system component that provides a printout of sample results and other information Prism Phenotype plot for multicolor analysis Prism histogram Histogram that displays the phenotype of an identified population Protocol A set of instructions that tells the Cytometer what and how to acquire data and relay listmode data Quality control QC A comprehensive set of procedures a laboratory sets up to ensure that an instrument is working accurately and precisely QuickCOMP Direct histogram manipulation to adjust Cytometer color compensation QuickSET Direct histogram manipulation to adjust Cytometer voltages and gain RD1 Abbreviation for phycoerythrin dye GLOSSARY 5 GLOSSARY Romlock A software licensing mechanism that is attached to the computer that enables access to the MXP software Runtime Protocol The Protocol stored with the listmode file at acquisition Listmode replays identical to the acquisition protocol Sample ID ID assignments made to a well or tube on the Plate Wizard to identify acquisition information Up to four Sample IDs can be defined per well or tube Scroll bar The area on the left of a pop up window The bar s arrows let you move scroll the window s content up or down so that you can see other parts of it For example the scroll bar in the Resource Exp
110. advanced precedence B 131 blend color B 128 boolean gating B 125 color blend mode B 127 gate and logical expression B 122 gate color B 124 gate logic B 125 gate name B 128 modify color blend B 128 modify color precedence B 130 operands B 123 histogram freeze frame B 103 histogram plot B 100 legend plot B 116 linear regions B 136 linked marker B 137 multiple linear regions B 137 new acquisition protocol B 12 new panel B 13 new plate B 14 new protocol B 11 new worklist B 14 non linked marker B 137 polygonal regions B 132 PRISM plot B 110 dividers B 113 dual parameter PRISM dividers B 113 B 143 phenotypes B 112 single parameter PRISM divider B 113 B 145 quadrant regions B 135 rectangular regions B 134 cross cylindrical lenses definition GLOSSARY 2 customize toolbars command tab B 73 toolbars tab B 72 cut edit menu B 59 CV definition GLOSSARY 1 Cyto Comp cells description 1 5 See controls Cytometer abort INDEX 6 abort the MPL B 172 abort the panel B 172 abort the tube B 172 repeat the panel B 173 repeat the tube B 173 cleanse B 173 control translucency acquisition options B 49 cytosettings get cytosettings from LMD file B 171 get cytosettings from protocol B 171 definition GLOSSARY 2 description 1 2 idle mode B 173 menu B 164 prime B 173 prime if idle too long B 173 restart B 173 serial number user info B 51 settings status page B 171 specifications 3
111. ages settings Cytometer Control B 170 voltage specific power supply dimensions 3 1 See also Power Supply W waste disposal 2 2 requirements 2 2 waste container capacity 2 2 connections 2 5 waste line connection 2 5 welcome to MXP software B 1 Well Loc acquisition manager B 68 LMD file name B 46 well location definition GLOSSARY 6 display in acquisition manager B 68 well number definition GLOSSARY 6 LMD file name B 45 window See pop up window window menu B 179 Windows arrange icons B 181 cascade B 179 close all B 181 force square B 181 Tile B 179 Tile Special B 180 use of security features C 3 wizard panel See panel wizard plate See plate wizard startup See startup wizard PN 177579BD INDEX WLQ file extension 3 6 WLS extension for worklist file 3 6 workbook MXP DaCs Interface Workbook C 5 See also Excel worklist columns available in acquisition manager B 65 columns reorder in acquisition manager B 65 file extension 3 6 print B 64 view B 64 See also acquisition manager worksheet labeled _Audit C 5 labeled _System C 5 labeled Result C 5 See also Excel workspace preferences acquisition options B 48 gating B 54 introduction B 44 LMD file name B 45 plot display B 51 publish B 56 user info B 50 Workstation definition GLOSSARY 7 description 1 2 server option 1 3 specifications 3 4 X X red X in startup wizard B 2 Z zoom B 73 INDEX 23
112. al We can repeat this with 3 4 or more parameters The Prism Divider can be used instead to mark the dividing line between the Negative and Positive cells Divider 1 Divider 2 Divider 3 Phenotype components Phenotype components are written from left to right as Divider 1 Divider 2 Divider 3 B 112 PN 177579BD USING MXP SOFTWARE B PLOTS MENU Prism Dividers A Prism is created by setting one or more prism dividers There are three ways of setting prism dividers two of them involve graphical positioning single and dual prism dividers The third uses Analysis gt gt View Modify Prism that can set the divider numerically on any divider Any changes to the divider on one plot is replicated on any other plot using that parameter Each new parameter that has a prism divider set on it multiplies the total number of phenotypes by 2 Only one divider can be set per parameter Setting a Single Parameter Prism Divider Single parameter prism dividers are set by xi placing a Single parameter histogram in focus and selecting The divider can then be set by clicking on the selected plot positioning the divider and then clicking once more to fix the divider position Dual Parameter Prism Dual parameter prism dividers are set by x placing a Dual parameter plot in focus and a selecting F This simultaneously creates two prism dividers
113. ample lines with cleaning agent The Cytometer must be in Idle mode E to perform this function Status Bar The Status Bar which is shown at the bottom of the screen displays the following information when in Acquisition Awaiting Sample m 0 05 21374 events Q lt No messages received gt A SERS 12 26 PM Cytometer Status Awaiting Sample Elapsed Acquisition Total Events Indication of Events Sec Time Secs Acquired See also Total Events Elapsed Time Data Rate PN 177579BD B 173 B USING MXP SOFTWARE CYTOMETER MENU Total Events The current total number of events acquired is displayed here When acquiring this value increments to the maximum events set or until one of the Stop conditions has been met See Acquisition Stop and Save When acquiring data in Setup Mode this item cycles in the range from 0 to 10 times the number of Events shown in the Dots Displayed The value appears to wraparound that is if you have requested 5 000 dots to be displayed this item increments to 4 999 On the 5 000th event the total events counter resets to 0 This ensures that you can continue to display data without reaching a set stop condition Elapsed Time This item shows how many seconds have elapsed since the start of data acquisition The total elapsed time can be viewed from the FCS Keywords associated with the beginning time and end time of acquisition Data Rate This item shows the speed of data acquisition
114. annel number yy channel value PnU Units for this parameter n PnQ Sensor name for parameter n from the CCUI Pnc Calculation method for parameter n ARITHMETIC means use arithmetic method for calculation may be overridden by analysis engine PnADDRESS Mutliplexor address of parameter n RUNNUMBER See SMNO System defined run number that is a number which is incremented each time the flow cytometer is used to acquire data BASELINEOFFSET Default is OFF any other means ON that is this sample was acquired using baseline offset FILEGUID Unique 128 bit number identifying this file changed every time the file is saved CYTOMETERID The unique ID given to the flow cytometer by the user for example the flow cytometer serial number LOCATION The address of the institute who owns the flow cytometer TESTNAME PROTOCOL NAME TESTFILE PROTOCOL FILE NAME BARCODE No Read CAROUSEL 1 TUBENO Well number corresponds to the number that appears within the well displayed on the Plate Wizard Example 01 02 96 PnDIVIDER Prism channel divider OxFFFF means OFF COMPENSATIONMODE Displays the method of compensation implemented Advanced or Classic Discriminator Displays the discriminator parameter and level REDLASER Displays the status of the Red Laser during analysis of the sample TARGETLASERPOWER Displays the Argon Laser power set during acquisition RESAVEDFILE Displa
115. appearing when the plot is printed or exported Show Axis Tick Marks Unchecking these options hides the axis scale tick marks and labels Show Axis Labels Unchecking these options allows the axis parameter name to be hidden X and Y Axis Labels Allows custom naming of the parameters on the current plot any FCS keyword or text may be entered Alternatively select Pick to select from File Name Stain name PnS or Parameter name PnN Font Sete Oe aed E pad one xi and Point Size Data Source Events Resolution Labeling Font Stop and Save Other font attributes available are Underline Strike Out and Text Color The default color is T Arial Black Tp Aral CE black doe Bald Italic T Arial Greek The Apply To All Plots checkbox allows the Bee is cee selected font to be applied to all the Tr Courier New 7 AaBbCcDd currently displayed plots You should Text Color choose True Type fonts so that X and Y axis i I Underline J Strike Out J Apply To All Plots displays are available Cancel Help PN 177579BD B 99 USING MXP SOFTWARE PLOTS MENU Histogram Plot Single parameter histograms show the frequency distribution of the chosen parameter All histograms can be customized that is Colors Gate for histogram Scaling and so on To create a Histogram Plot Og 2 From the Data Source dialog box x displayed select the required File then caa cre ae select
116. are logged in Password Enter your User Password Passwords are a minimum of 6 characters and a maximum of 24 characters Blanks are not allowed For best security create a password with at least eight characters including letters numbers 0 9 and or symbols If you exceed the number of unsuccessful password attempts allowed per the System Administrator you will not be allowed to log into the MXP software You must then ask your System Administrator to unlock you and or reset your password before you can log into MXP software The system prompts you with a Password No Longer Valid message whenever you must change your password You must change your password e The first time you log into MXP software after the software was installed e The first time you log into MXP software after the System Administrator has reset your password Each time your password expires Your System Administrator sets up the time interval when you must change your password Use the Password Change dialog box that appears to enter and confirm your new password Enter New Password e O Admin Button Allows the System Administrator access to the User Administration Tools Finish Button Starts up MXP software after you select your User ID and enter your password B 3 B USING MXP SOFTWARE TOOLBAR BUTTONS B 4 B 4 TOOLBAR BUTTONS Toolbars form an essential part of the MXP software The application entered and the mode you are
117. arithmic amplification A method of increasing the gain and dynamic range of a signal A larger gain is applied to a sensor s smaller signals than to the sensor s larger signals See also gain MB Abbreviation for megabyte Mean Arithmetic average of a group of data See also standard deviation and coefficient of variation Menu On a Workstation screen a list of items from which you can choose Minimum Event Counter The count that must minimally be achieved in order to stop acquisition Used to ensure collection of rare events Mouse A pointing device The cursor on the Workstation screen moves as you slide the mouse on your desk or other flat surface Multi Plate Loader MPL An automated sample loader for the instrument Neutral density ND1 filter An optical filter that can be used with the forward scatter sensor to reduce the intensity of the forward scatter thus enabling the instrument to analyze large particles without saturating the sensor Optical filters Mediums such as glass that separate fluorescent light by wavelength which is measured in nanometers nm See also BK BP and DL filters Panel In general a group of protocols for analyzing a series of samples corresponding to one specimen The Cytometer settings and regions can be passed on through the panel with identification of the primary samples GLOSSARY 4 PN 177579BD PN 177579BD GLOSSARY Panel Wizard A software feature that guide
118. art 11 option and the spreadsheet is under DaCS software control when the data acquisition is done go to the Excel software and save the spreadsheet generated W DaCS gt gt Save Workbook m PN 177579BD USING MXP SOFTWARE FILE MENU 19 Select Tools gt MPL_DaCSAnalysis or E7 speling F7 Tools MPLAnalysis as needed Ba Error Checking MPL_DaCSAnalysis Share Workbook Protection Online Collaboration gt Formula Auditing Tools on the Web Add Ins Customize Options y Y Spelling F7 Error Checking MPLAnalysis Share Workbook Protection gt Online Collaboration gt Formula Auditing gt Tools on the Web Add Ins Customize Options y 20 A screen message prompts you to open the workbook file Use the Browse box that appears to select the xls file PN 177579BD B 21 USING MXP SOFTWARE FILE MENU 21 Review the graphs and data columns for the sample plate Values above the threshold value appear red in the data columns and a corresponding red threshold line appears on the graph You can modify the threshold value Note Each sheet name is made up of the region name followed by the statistic name B T EB H EJI Eol fs 500 16 450 Hw 9 350 10 Era 300 12 250
119. as been selected and not to the ones previously created Smooth Histograms This automatically smooths single histograms by default allowing for easier viewing When selected this option displays smoothed data on all new histograms as they are created It does not apply any of the changes to previously created plots Note If peaks observed during acquisition in a single parameter histogram disappear when the stop count is reached remove the check from the Smooth checkbox This condition occurs due to a small number of events in this plot PN 177579BD PN 177579BD USING MXP SOFTWARE FILE MENU Default Overlay Plot Background Color This feature is not available in MXP software Default of Events on Dot Plot This allows you to set the default number of dots to be displayed when creating a dot plot These changes are only applied to plots created after this option has been selected and not to the ones previously created Reducing the default percentage increases the speed of software functions when reading large listmode files Baseline Offset IMPORTANT Risk of erroneous results The Baseline Offset function should only be used after first viewing data with the baseline offset function turned Off unchecked You must be satisfied that the overall results of an assay are not significantly affected by turning baseline offset on If the lower marker of a statistics region is higher than the first decade of log displayed data the
120. ate Repeat the Well Repeat the Panel Ho Abort the Well Stops acquisition and marks the current sample as aborted The instrument proceeds to the next sample Abort the Panel Stops acquisition and marks the remainder of the current panel as aborted The instrument proceeds to the first sample of the next panel Abort the Plate Ejects the current sample plate or tube holder and leaves the remainder of the Worklist unread allowing resumption from the current sample Note If you select Abort the Plate in rare instances the sample probe may move to the wash station and the sample tray exits the instrument When this occurs you need to O TWICE so the system performs an initialization which leaves the system ready to acquire Repeat the Well Allows the current well to be resampled PN 177579BD USING MXP SOFTWARE CYTOMETER MENU Repeat the Panel Allows the current panel to be resampled The MPL returns to the first sample of the current panel Restart If a sample is already being acquired the Restart button a restarts the Acquisition from zero events Idle Mode The Idle Mode button e places the Cytometer in Idle mode so that various cleaning and replacing procedures can be performed Prime The Prime button allows you to declog or remove bubbles or blockages from the sample line with sheath solution Cleanse The Cleanse button allows you to initiate a cycle to flush the instrument s
121. ate To make parallel for example collimate rays of light Color compensation The subtraction of e a percentage of the signal from one fluorescence light sensor from e the signal from another fluorescence light sensor to correct for the overlap of one dye s emission into another dye s emission measurement Control A substance used to routinely monitor the performance of an analytical process that does not have the characteristic being measured for example Immuno Trol cells or CYTO TROL control cells Controls and indicators Instrument controls are the mechanisms you use to communicate with the instrument Indicators are the mechanisms the instrument uses to communicate with you Cross cylindrical lenses Used in the Cytometer to focus the laser beam and form an elliptical beam spot Cytometer The system component that analyzes the sample and contains the sheath fluid and cleaning agent bottles Cytosettings Cytometer hardware settings Consists of acquisition duration acquisition maximum events gating settings disciminators voltages gains baseline offset and compensation settings Same as Instrument Settings in Acquisition Manager db Abbreviation for decibels dc Abbreviation for direct current Defaults Original settings for the instrument You can change them to customize the settings for your laboratory DiOC5 3 Abbreviation for oxacarbocyanine dye Discriminator A channel setting for a pa
122. ate around the selected population on a plot Create a Contour AutoGate around the selected population on a plot Create or Edit a Dual Parameter plot with a set of Dual Prism Dividers Insert a Linear Region Insert Multiple Linear Regions Create a Single Parameter Prism Divider GATE COLOR STATS AND HELP TOOLBAR e o s Ce e E Create Modify Gate Modify Color Gate Blend Setup Create Modify Color Gate Precedence Setup View Current Regions Publish Results Select Results Help Beckman Coulter ACS Access the Create Modify Gates dialog box Access the Modify Color Gate Blend Setup dialog box Access the Create Modify Color Gate Precedence Setup dialog box View edit a Region s properties Region points or to delete a Region that is no longer required Publish the statistics to an MS Excel spreadsheet or a Text file depending on your Workspace Preferences selection Choose Statistics for display and export Access the Help System Go to the Beckman Coulter Inc web site Go to the Applied Cytometry Systems web site PN 177579BD B 9 FLOWPAGE TOOLBAR A New FlowPAGE Insert Bit Map Annotation Line Rectangle Align Top Align Bottom HEH OWES oe Align Left ee Align Right Space Evenly Across Space Evenly Down Grow to Largest Shrink to Smallest A ee EB USING MXP SOFTWARE FLOWPAGE TOOLBAR Open a New FlowPAGE Also see Insert Blank FlowPAGE Insert a
123. ater than sign gt less than sign lt asterisk comma question mark quotation mark pipe symbol colon or apostrophe Leading trailing and multiple spaces are not allowed peat Ore xl Sample ID 1 2 3 4 Plot Display Gating Publish Well Loc LMD File Name Acquisition Options User Info User 1D m Listmode File Name Run Number J First 10 characters of protocol name Y2K Date I Sample ID 1 V Run Number Time I Sample ID 2 I 2K Date YYYY MM DD Tag Number I Sample ID 3 I Time HH MM File Name Extension J Sample ID 4 V Tag Number PDF File Name Extension F well Loc C Start each panel at 001 Next File Name UserID Start At 001 Note If more than one Listmode File m Filename Extension PDF Filename Extension Name is selected the priority of the Teg Nate G Append PDF options used for the file name are LMD C Replace 1 Well Loc Next File Name 2 Sample ID 1 3 Protocol ID 00000000 001 LMD 4 Sample ID 2 5 Sample ID 3 6 Sample ID 4 oe a e toon e 7 Run Number 8 Tag Number 9 User ID 10 Date 11 Time See also Workspace Preferences Introduction Workspace Preferences Acquisition Options Workspace Preferences User Info Workspace Preferences Plot Display Workspace Preferences Gating Workspace Preferences Publish B 45 B USING MXP SOFTWARE FILE MENU B 46 First 10 charac
124. ave all B 31 header edit attributes B 41 keywords 3 9 legend plot B 118 view information B 40 information legend plot B 118 select results B 149 keyword list 3 9 save histogram data in FCS format B 121 standard keywords 3 9 FDA definition GLOSSARY 2 file display signed ones B 25 display versions B 24 export definition GLOSSARY 2 histogram file save as B 32 how to check in uncontrolled copy B 71 menu B 11 open histogram B 30 open listmode B 23 open protocol B 34 options toolbar B 5 plot data source B 96 protocol save as B 38 references in data B 56 save all FCS format files B 31 save protocol B 37 sign version B 177 uncontrolled file B 71 file name extension LMD file name B 47 file references in data B 56 specifications 3 6 unavailable characters 3 6 filter block dual laser 1 3 single laser 1 3 filter block dual laser options 1 3 filter block single laser options 1 3 filter kit dual laser 1 3 options 1 3 single laser 1 3 INDEX 9 INDEX filter kit dual laser options 1 3 filter kit single laser options 1 3 filters BP definition GLOSSARY 1 contained in optional filter block 1 3 contained in optional kit 1 3 DL definition GLOSSARY 2 dual laser filter block optional 1 3 dual laser filter kit optional 1 3 filter kit optional 1 3 ND1 definition GLOSSARY 4 optical definition GLOSSARY 4 optical list of 3 2 optical specifications 3 2 single laser fi
125. blish your application settings AUX Signal Auxiliary acquisition pathway that allows either control of simultaneous Lin and Log signals or acquisition of a Peak signal Background count Measure of the amount of electrical or particle interference BCAQG P database This is the Beckman Coulter Accounts amp Permissions database in the MXP software BP filter A band pass optical filter that passes a band of wavelengths and blocks others Button The Workstation screens have pictures icons for example that you select with the mouse to tell the software what to do They are arranged on Toolbars for related functions Cal Factor A number used in conjunction with a known number of particles identified by a CAL region that adjusts the region counts obtained Channel In an analog to digital converter the number of equally spaced divisions of the amplified input signal voltage All FC 500 MPL flow cytometer signals are resolved into 1024 channels For dual parameter histograms the number of channels is reduced to 64 128 or 256 Cleaning agent A detergent used to flush sample from tubing and minimize protein buildup Click To press and release a mouse button Coefficient of variation CV A measure of the variability in signal intensity that is generated as particles pass repeatedly through the laser beam This variability is expressed as a percentage of the average signal intensity GLOSSARY 1 GLOSSARY Collim
126. button x and 3 only the user or the System Administrator can use the File gt Log In feature to return to the MXP software These are the automated activities gt Acquisition gt Listmode playback gt Batch AutoMATOR gt AutoSetup Manual Lock Out The user can manually lock the MXP software all of the features are unavailable with the Lock Software feature of the 21 CFR 11 drop down menu The user should lock the software whenever the software will be out of the user s control The user or System Administrator must use the File gt Log In feature to return to the MXP software PN 177579BD 21 CFR PART 11 OPTION COMPLIANCE INFORMATION INTEGRITY OF ELECTRONIC DATA MXP Software Files MXP software ensures listmode LMD histogram protocol panel and PDF file integrity Whenever one of these file types is created by MXP software or checked into MXP software it is assigned a version number Each time a change is made to one of these file types an incremental version number is assigned when the file is saved a comment is required to be saved with each version to describe the action done to the file During automated activities automatic system generated comments are saved to the files otherwise the user must enter a comment when saving a new version of a file Uncontrolled Files that exist in Windows are not under the control of the 21 CFR Part 11 option and cannot be used by the MXP software until they are checked in
127. cation is loaded into a directory labeled C MXP The MXP software also contains the Beckman Coulter Accounts and Permissions BCA P database which uses Microsoft SQL Server 2000 These items are installed as part of the MXP software version 2 0 installation procedure Microsoft Excel Software Excel software is provided for exporting data Only Microsoft Excel versions 2002 and version 2003 have been tested to be compatible with the MXP software No other version of Excel software should be used 1 2 PN 177579BD USE AND FUNCTION OPTIONS 1 2 OPTIONS Hardware Options Printer Provides a printout of sample results and other information The Printers available for the system are subject to change so contact your Beckman Coulter Representative for the current model selection Workstation Network Server A Beckman Coulter FlowCentre Multimedia Workstation can be used as an offline analysis workstation and network server for data and worklist transfer Second Laser A 635 638 nm red solid state laser providing a minimum of 25 mW light regulated laser power The second laser has a shutter that allows you to block the laser output Filter Kit The optional filter kit includes 5 extra filter holders an ND 1 0 filter a full mirror in a holder a 50 mirror in a holder and an additional filter block base Single Laser Filter Block An assembled filter block configured as shown in Figure 5 7 in the Instructions For U
128. cess http www beckmancoulter com 1 4 MATERIAL SAFETY DATA SHEETS MSDS To obtain an MSDS for Beckman Coulter reagents used on the Cytomics FC 500 MPL flow cytometer l 2 PN 177579BD On the internet go to http www beckmancoulter com a Select MSDS from the Customer Support drop down menu b Follow the instructions on the screen c Contact your Beckman Coulter Representative if you have difficulty locating the information If you do not have Internet access e Inthe USA either call Beckman Coulter Customer Operations 800 526 7694 or write to Beckman Coulter Inc Attention MSDS Requests PO BOX 169015 Miami FL 33116 9015 e Outside the USA contact your Beckman Coulter Representative 1 5 i USE AND FUNCTION MATERIAL SAFETY DATA SHEETS MSDS 1 6 PN 177579BD INSTALLATION 2 1 DELIVERY INSPECTION The instrument or MPL upgrade kit is tested before shipping International symbols and special handling instructions are printed on the shipping cartons to inform the carrier of the precautions and care applicable to electronic instruments CAUTION Possible instrument damage could occur if you uncrate the instrument install it or set it up Keep the instrument in its packaging until your Beckman Coulter Representative uncrates it for installation and setup When you receive your instrument carefully inspect all cartons If you see signs of mishandling or damage file a claim with the carri
129. configuration availability Computer Intel Pentium 4 microprocessor with a minimum of 256 MB of RAM Data Storage e 3 5 in 1 44 MB diskette drive e 20 GB minimum nonremovable hard disk e 700 MB CD RW drive e 250 MB minimum internal ZIP drive Interfaces e Windows 2000 or Windows XP e Bidirectional asynchronous serial interfaces for communication with mainframe and personal computers Input Devices e PS 2 optical Intellimouse e Full size PS 2 keyboard Monitor Options Color LCD with a 17 in flat screen PN 177579BD 3 3 PN 177579BD SPECIFICATIONS SOFTWARE SPECIFICATIONS SOFTWARE SPECIFICATIONS Data Output and Compatibility Output and compatibility features include Flow Cytometric Standard FCS file format for listmode and histogram files Listmode files contain an FCS 2 0 dataset followed by an FCS 3 0 dataset Copy and Paste or Drag and Drop plots images to third party programs such as MS Word Full color printouts with the appropriate optional Printer Printout of sample results with FlowPAGE and desktop printing PDF format for Acquisition or FlowPAGE printouts Printout of Worklist for the MPL Export data to MS Excel Export billing information to MS Excel Save export data as a text file Setup Mode During Setup mode the workspace is a rolling display that is updated up to three times per second The maximum number of events can be set on the Cytometer Control Acquisiti
130. connections 2 5 description 1 4 sheath fluid definition GLOSSARY 6 types used 1 4 See also cleaning agent sheath fluid rectangular region creating B 134 interactive information B 135 properties B 134 red laser disable B 168 enable B 168 See also laser and HeNe or solid state laser PN 177579BD redo edit menu B 59 reference manual conventions used xxii introduction to your manuals xxi using the manuals xxi refresh B 73 regions autogating B 135 contour sensitivity B 55 contour travel B 55 elliptical sensitivity B 55 autosetup target region B 136 carry from protocol B 66 carry previous tube B 66 creating autogating region B 135 linear region B 136 multiple linear regions B 137 polygonal region B 132 quadrant region B 135 rectangular region B 134 display sources in acquisition manager B 66 editing B 138 delete region B 141 move region B 139 region name B 140 region points B 139 resize region B 139 select region B 138 introduction B 131 linear region B 136 interactive information B 136 properties B 136 load B 66 maximum per protocol 3 5 multiple linear regions B 137 interactive information B 137 plot data source B 97 polygonal region B 132 interactive information B 134 properties B 132 properties format tab B 134 prime B 136 properties B 132 linear region properties B 136 polygonal region properties B 132 quadrant region properties B 135 rectangular region pr
131. copies as needed PN 177579BD A 1 LOG SHEETS A 2 PN 177579BD LOG SHEETS A REAGENT LOG Concentration Date Date Expiration R tN ts eagent Name Units Received Opened Lot Number Date Tech If applicable Cytometer Serial No A y Cj BECKMAN Laboratory COULTER Cytomics FC 500 MPL 7579022A A 3 PN 177579BD LOG SHEETS A 4 PN 177579BD ACTION LOG LOG SHEETS A Date Condition Noted Tech Date Action Taken Tech Cytometer Serial No Laboratory Cytomics FC 500 MPL PN 177579BD Saecos COULTER 7579023A A 5 LOG SHEETS A 6 PN 177579BD LOG SHEETS A DAILY LOG FOR INSTRUMENT STANDARDIZATION Application Flow Set Fluorospheres Lot Number Expiration Date Flow Set 675 Fluorospheres Lot Number __ Expiration Date Flow Set 770 Fluorospheres Lot Number Expiration Date Instrument HV Total Gain Target Ranges FS LOG FL1 LOG FL3 SS LOG SS LOGFL2 LOG FL4 LOG FL5 FS SS or LOG SS LOG FL1 LOG FL2 LOG FL3 LOG FL4 LOG FL5 Red laser 488 Tech Run Mode HV Gain Mode HV Gain Mode HV Mode HV Mode HV Mode HV Date
132. ct 2004 17 17 08 Signature Certified Signed by Mr Anderson Signed on 06 Oct 2004 17 17 23 Signature Authorized Signed by Mr Anderson Signed on 06 Oct 2004 17 17 38 z 8 At any time you can D to hide the Checked In Versions portion of the screen B 25 B USING MXP SOFTWARE FILE MENU 9 To open the file you want highlight the file name and either D to open the most recent version of the selected file OR e Highlight a specific version of the file and D to open that version Note If you selected an archived file that has not been restored a message appears stating the archive location of the file Supply the file location to your System Administrator and request the file be restored When loading listmode files which have a parameter order different from the current protocol you are warned with Parameter Mismatch e Ifyou select Continue the file is loaded and replaces the parameters with those of the listmode file e Ifyou select Abort the listmode file is not loaded Note Listmode Compensation must be performed using the runtime protocol or a protocol with equivalent parameters as the runtime protocol When loading listmode files from a different instrument This is not an FC 500 file appears Select Continue to load the file B 26 PN 177579BD USING MXP SOFTWARE FILE MENU Open Listmode File Procedure Standard MXP Software Installed Use this option to open a new listmode file into al
133. ctive Linear Region Editing Region Properties x Region Properties Format Region Name Lymphs J Prime J7 Automatic region name positioning Autogating Hone Sensitivity fir gt Elliptical Ory Travel E Minimum Count Ir Active Count E Cancel Help PN 177579BD PN 177579BD USING MXP SOFTWARE ANALYSIS MENU Region Name Displays the name of the region If the name is not grayed you can rename it Prime When checked this option allows the system to determine if the peak of the events on the plot is within this region If the peak is outside this region due to an air bubble or partial clog the system performs a Prime cycle A maximum of three sequential Prime cycles are allowed if the peak of the plot is still not within this region the MPL aborts Automatic Region Name Positioning Region labels default to an automatic position You may move a region label to any desired position within a plot this user modification automatically unchecks this option To revert to the automatic position recheck this option AutoGating None This is the default when the AutoGate option is not selected Elliptical Toggle the AutoGate to an elliptical region Contour Toggle the AutoGate region to a contour region Sensitivity Eliptical AutoGate settings are 0 11 0 33 and 1 1 of processed peak Contour AutoGate settings are 1 2 3 4 and 5 e Level 1 0 11 processed peak at a resolution of 64
134. currently on the desktop Acquisition Additional steps are needed to publish to Excel during acquisition and are noted in this procedure Ensure the MPL_DaCSAnalysis Add In or MPLAnalysis Add In is installed in Excel e C Tools in Excel e The appropriate Add In MPL_DaCSAnalysis or MPLAnalysis appears on the Tools menu ae Spelling F7 Error Checking MPL_DaCSAnalysis Share Workbook Protection gt Online Collaboration gt Formula Auditing Tools on the Web Add Ins Customize Options v lt Spelling F Error Checking MPLAnalysis Share Workbook Protection gt Online Collaboration gt E Auditing Tools on the Web Add Ins Customize Options y PN 177579BD USING MXP SOFTWARE B FILE MENU 2 In the MXP software select File gt gt Workspace Preferences W x LMD File Name Acquisition Options User Info 3 the Gating tab Plot Display Gating Publish Dot Plot Coloring Mode M Automatic Gate Creation Color Blend Mode Color Precedence Mode M Automatic Gate Maintenance M Automatic Color Precedence 4 O Iw Automatic Gate Maintenance C eiti Plot Display Gating Publish 5 the Acquisition Options tab OFENSA Acquistion Options Userinfo Output Options M Save LMD T Print Plots T Publish I Save Histograms JT Pri
135. d as you move the mouse to either the left or right of this point the integral value continuously updates to reflect the summation of each channel between the Region starting position and the current mouse position Create Multiple Linear Regions This option sets multiple consecutive linear Regions on one display See Create Multiple Linear Regions in the Instructions For Use manual Interactive Multi Linear Region Editing When a Region s position is changed any Gates that use the Region are automatically updated and each plot is redrawn to reflect the changes in the Gate When the Region is deleted any Gate which used the Region is also automatically deleted When a Gate has been deleted any plot that used the deleted Gate is reset to Ungated Allows you to resize an already created Region Linked Marker This feature is not available in MXP software New Non Linked Marker This feature is not available in MXP software PN 177579BD B 137 USING MXP SOFTWARE ANALYSIS MENU B 138 View Modify Regions Use this option to view edit a Region s properties Region points or to delete a Region that is no longer required Allows you to copy and paste Regions between protocols Os Make the desired changes x See also Regions Lymphs 8 CD45 Lymphs Cancel Region Points Delete Help Region Points 3 O or press to accept the changes D or press to exit the dialog box without saving the changes S
136. d the button is active B 32 PN 177579BD USING MXP SOFTWARE B FILE MENU 5 W to save the file MXP software creates a file with the name and with the version and comment if applicable that you specified and stores it in the drive and directory you specified in FCS histogram format Save Histogram File As Procedure Standard MXP Software Installed Use this procedure if you have the standard MXP software installed without the 21 CFR Part 11 option 1 Highlight the Histogram Plot Current window 2 J File Save Histogram File As 3 Specify the name of the file aixi Save in Sy HST 7 ef EB a 2c CDS CD19 Lymph FLI Log HST a 2c CDS CDS Lymph FL1 Log HST a 2c CD3 CD19 Lymph FL2 Log HST se 2c CD3 CDB Lymph FL2 Log HST a 2c CD3 CD4 Lymph FL1 Log HST a 2c Neg Control Lymph FL1 Log HST fa 2c CD3 CD4 Lymph FL2 Log HST a 2c Neg Control Lymph FL2 Log HST i 2c CD3 CD56 Lymph FL1 Log HST a 2c CD3 CD56 Lymph FL2 Log HST a the desired directory in the Save in list box b D the type of file in the Save as type list box Filename 00000628 113 Ungated FL1 Log HST c In the File name text box type the Save astype HST Fies hs eel desired File Name 4 W to save the file MXP software creates a file with the name that you specified and stores it in the drive and directory you specified in FCS histogram format PN 177579BD B 33 USING MXP SOFTWARE F
137. der polynomial fit It is used to p mat gee gt gt determine the concentration 2 equivalent values for the ee e E a e A separate column is added to the remaining Mean Median and Mode sheets with the calculated concentration equivalents for sample wells with their Well Type designated as Specimen in the Plate Wizard B 22 PN 177579BD USING MXP SOFTWARE B FILE MENU 23 Save the graphs and data spreadsheet D the selections appropriate to the Analysis Add In used gt For the MPL_DaCSAnalysis Add In use DaCS gt Save Workbook gt For the MPLAnalysis Add In use File gt gt Save As e Enter anew or the same xls file name Caza Open Listmode File Use the open listmode feature to open a new listmode file into all plots Before any analysis can be done a listmode file must be read into MXP software Most buttons on the Button Bar are disabled until this has been done Use the appropriate procedure depending upon whether you have the standard MXP software or also have the 21 CFR Part 11 option installed Select the appropriate link e Open Listmode File Procedure With 21 CFR Part 11 Option Installed e Open Listmode File Procedure Standard MXP Software Installed Open Listmode File Procedure With 21 CFR Part 11 Option Installed Use this procedure if you have the 21 CFR Part 11 option installed on your system Ask your System Administrator to restore
138. dering Smooth If the Smooth box is checked the visual representation of the Histogram is smoothed Note If peaks observed during acquisition in a single parameter histogram disappear when the stop count is reached remove the check from the Smooth checkbox This condition occurs due to a small number of events in this plot Freeze Frame Check a Freeze Frame box to assign the current Histogram The top button creates a dotted overlay and the bottom button creates a dashed Histogram The current Histogram is assigned to whichever Overlay button is selected and its name appears in the edit box adjacent to the button This legend may be modified if desired In order for Freeze Frame to work in Acquisition Manager ensure that you select Use Plots and Gates from Previous Test in the Panel Wizard for the panel Histogram Freeze Frame MXP software allows the display of up to three Histograms on each single parameter Histogram The base Histogram is the one that relates to the current listmode file and the other two can be overlays from any other compatible file Statistics are calculated from the base Histogram only The other two overlays are for comparison only and no statistical information can be obtained from them See also Create a Histogram Freeze Frame Delete a Histogram Freeze Frame Overlay PN 177579BD PN 177579BD USING MXP SOFTWARE PLOTS MENU Create a Histogram Freeze Frame In order for Freeze Frame to work i
139. e E Heed Save in list box l R as 3c 11 settings pro BAAS ACIL H B as 3c 11 flow set pro BAS FLIC p File name enpo o Save b O the type of file in the Save as a type list box Saving As Version 4 i Default Comments MEE a c In the File name text box type the cee a ie desired File Name printed PDFs PN 177579BD B 38 PN 177579BD USING MXP SOFTWARE FILE MENU You must enter a comment before you Can Save a New version e Select an appropriate comment from the list of Default Comments OR e Type in a comment in the Comments for this version field Note Once a comment is entered the version number automatically appears in the Saving As Version field and the button is active W to save the file MXP software creates a file with the name and with the version and comment if applicable that you specified and stores it in the drive and directory you specified in FCS format MXP software gives the default file extension PRO The PRO extension is only a suggested extension any standard DOS characters can be used for the full File Name You are urged to keep the extension consistent for easy file searches Save Protocol As Procedure Standard MXP Software Installed Use this procedure if you have the standard MXP software installed without the 21 CFR Part 11 option 1 Sharlene Save Protocol 2 xI Save in SJ AcquisitionProtocol e et Fae a AS 2C Settings PRO a AS 4C
140. e a different parameter name for each detector Parameter Selection x m Detectors N ln Li r Ratio Selected Signals 1E tore Numerator FS Lin he ie E u m Denominator 55 Lin X s ss K E Derived Parameters FL1 Fu C KM Time E Ratio E FL2 F2 goa FL3 FL3 rw FL4 F4 goa FES FL5 r iW Ausiliary Parameters 7 FS x aux C i PN 177579BD USING MXP SOFTWARE CYTOMETER MENU You can arrange the order within the Selected Signals list box by highlighting the appropriate item s and Drag and Drop into your chosen order within the Selected Signals list box To remove a previously selected item from the Selected Signals box uncheck the parameter no longer required Ratio and Time cannot be reordered and always appear at the bottom of the list Ratio Choose from the drop down box the appropriate option for the Numerator and Denominator if required A detector name may be entered this is displayed in all Cytometer Control screens but does not appear on plots Cytometer Control Detector Tab PN 177579BD This shows the detectors with x editable names and detector settings Acq Setup Detectors Compensation Changes made here are only reflected in the Cytometer Control dialog box and are used to identify the various fluorochromes in use Reagent or sample specific information should not be entered in this menu See View gt gt Acquisition Manager gt Parameter Names or File Edit FCS H
141. e keyboard _ indicates a key such as Enter indicates that the two keys listed such as Att F2 are linked for a specific function and must be pressed in this sequence a Press down on the first key listed and while continuing to press it press down on the second key listed b Release both keys at the same time _ indicates to press and release the first key listed then press and release the next key listed For example Enter Icons buttons to select functions on the software screen are shown within text Example O ee C indicates to use the mouse to select the screen button labeled D File gt Save indicates to use the mouse to select the Save item on the File menu through are special function keys A Note contains information that is important to remember or helpful in performing a procedure The terms screen and window are used interchangeably SHOW ME means there is a video available for the procedure in the online help PN 177579BD INTRODUCTION GRAPHICS To Choose A Command With The Keyboard After you press alt each menu name has one letter underlined to indicate which letter to use to pull down the menu For example the letter F in the File menu is underlined press FJ to pull down the File menu the letter E in the Edit menu is underlined press E to pull down the Edit menu Command Function Enter Accepts your selection Eso Stops the operation discard
142. e name Note The Listmode File Name column displays only a provisional file name prior to acquisition If Time or Run Number is included as part of the name the actual value of these fields can only be determined once acquisition of each sample is complete Therefore if you wish to print a worklist including the final listmode file names only do so once acquisition of all samples in the Worklist is complete PN 177579BD USING MXP SOFTWARE B FILE MENU Tag Number This allows you to choose whether the Tag number should be included as part of the listmode file name If this option is selected it activates the following items e Start each Panel at 001 This starts the Tag Numbering option from 001 for each new panel e Start At This starts the Tag Numbering option from any number that you require and continues incrementing until manually reset File Name Extension This sets the file extension to be either Tag Number or LMD e Tag Number Selecting this option allows you to have the Tag Number as the file name extension This is dependent on a Tag Number being previously selected e LMD This option sets the file name extension to be LMD PDF File Name Extension Change the LMD File Name extension to one of the options listed below Append PDF File Name Append PDF to the listmode file name extension Replace Replace the current listmode file name extension with PDE This option should only be used when LMD is being used a
143. eader Attributes Standard Detectors Auziliary Parameters Name Volts Gain AUX wo See also Cytometer Control Cytometer Control Acquisition Setup Tab Cytometer Control Compensation Tab Cytometer Control Parameter Setup Name These input fields allow the editing of the Names assigned to the default manufacturer s detectors Changes are ONLY reflected in the names shown within the Discriminator box in the Acquisition Setup tab and the Compensation tab This allows the various fluorochromes to be easily recognized when adjusting instrument settings B 169 B USING MXP SOFTWARE CYTOMETER MENU B 170 Volts Select the appropriate Voltage for the particular Detector This is done by using the Slide Bar on the right side of the screen and adjusting it to the required value For fine adjustment the Up and Down arrows at the ends of the Slide Bar can be used Gain Adjusts the Gain amplifier Set the Gain to the appropriate setting you require for that Detector This is chosen from the drop down box which appears when you click in the box AUX Displays the signal source for the Auxiliary Parameter channel Cytometer Control Compensation Tab Compensation allows for the E subtraction of a percentage of the Acq Setup Detectors Compensation signal of one fluorescence detector Compensation Matrix from the signal of another
144. ect a plot type b the correct parameters for the plot and 0 Lo c Right mouse click on the plot and D Format Plot 5 Modify or create subsequent plots and choose the required File Parameters Regions and any gates to be applied OB statistics output required Om 7 J Save Protocol As and enter a protocol name New Panel Use the Panel Wizard to create or edit a Panel for use in the Plate Wizard See Creating Panels or Editing Panels in the System Overview chapter in the Instructions For Use manual PN 177579BD B 13 USING MXP SOFTWARE FILE MENU New Plate Note If no Worklist is visible O View gt Acquisition Manager to display the Worklist pane W File gt New gt New Plate to open the Plate Wizard The main MXP screen automatically minimizes when the Plate Wizard opens See Creating Worklists in the System Overview chapter in the Instructions For Use manual for detailed information how to program a new sample plate Fie Help Acquistion Protocols Panels Protocol Name as 3c 11 settings pro as 3c TIflow set pro as 3c 1I neg contiol pro as 3c 1l very po as 4c 1I settings pro as 4c 1 flow set pro as 4c 1I neg contiol pro as do 1 verily pro as 4c 21 settings pro as de 2iflow set pro as 4c 2I neg control pro as 4c 2 ver po as 5c 11 settings pro as Se 1I flow set pro Well Information as 5c 1I neg control p
145. ee also Deleting Regions Editing and Moving a Region Name Moving a Region Resizing a Region Regions Select the Region to view edit or delete from the Regions list box Highlight the Region Name and B __Detete to Delete the region s Double click the Region Name to rename the region PN 177579BD USING MXP SOFTWARE ANALYSIS MENU Region Points This list box gives details of the Region bounds This is Upper Lower channel for single parameter histograms For rectangles the lower left and upper right vertices are specified and for polygons the X Y coordinates of all the vertices To display a Region s points or bounds click on the desired Region in the Regions list box Resizing a Region If you want to enlarge or shrink a Region 1 wW the Region The Region is now active and handles appear Vertices also appear for Contour Autogate and Polygon Regions 2 Hold the left mouse button down while moving the handle or moving a vertex using the cross hair cursor as in Contour Autogate and Polygon Regions Moving a Region To move a Region to another position 1 Mouse over the Region to get the th cursor 2 Hold the left mouse button down while moving the Region anywhere in the current window If the Region has been moved the statistics automatically recalculate The Region name moves to the same relative position alongside the Region s new position See also View Modify Regions PN 177579BD B 139
146. eferences Plot Display Dot Plot Data Source Font PN 177579BD B 97 USING MXP SOFTWARE PLOTS MENU No of Events to Show Place the cursor on the Slide bar and move to change the No of Events to Show Apply to All Plots Enable this checkbox to apply the No of Events to Show to all Plots Dot Plot Resolution Set the number of channels for the X and Y Axis of the Plot The available options allow for 64 128 256 xi and 512 channels to be displayed on the e a ao axis Data Source Events Resolution Resolution of plot A higher resolution gives an improved graphic image whereas a lower resolution allows faster redrawing of that image Note If a higher resolution is selected a sufficient number of events is required to produce a visually acceptable graph Cancel Help Dot Plot Labeling The options here allow you to override the default plot display options for the current plot This is useful when capturing images for publication arar That eon Cata xl Show Axis Tick Marks Data Source Events Resolution Show Axis Labels Labeling Font Stop and Save X and Y Axis Labels IV Show plot title on output IV Show X axis tick marks V Show axis labels IV Show Y axis tick marks V Show Y axis labels gt axis label STAN Pick v Y axis label ssTaIn Pick B 98 PN 177579BD USING MXP SOFTWARE B PLOTS MENU Show Plot Title on Output Unchecking this option prevents the plot title from
147. ent version of the file Only controlled files can be used in the MXP software with the 21 CFR Part 11 option Uncontrolled Files A non versioned file that is actually located in Windows and has not been checked into the MXP software s BCA P database Because uncontrolled files can be changed outside of the MXP software without a record of the change made they are not within 21 CFR Part 11 compliance To use an uncontrolled Panel Protocol or Listmode file you need to check it into the MXP software and the the BCAG P database How to Check In A File To check in uncontrolled copies of Panel Protocol or Listmode files into the BCA amp P database of the MXP software use any one of these methods e Drag and drop a protocol or listmode file from the Resource Explorer into your Workspace e Use File gt gt Open Protocol or File Open Listmode to open and check in a protocol or listmode file Select and run a protocol in the Plate Wizard e Select and run a panel in the Plate Wizard When you run a panel all of its protocols are also checked in Any Panel Protocol and Listmode files generated by the user within MXP software are controlled files receive a version number and do not need to be checked in Note Version numbers appear on Listmode and Protocol file names displayed in the lower portion of the Resource Explorer PN 177579BD B 71 B USING MXP SOFTWARE VIEW MENU B 72 Toolbars Customize Toolbars
148. er immediately If separately insured file the claim with the insurance company 2 2 SPECIAL REQUIREMENTS Before your Beckman Coulter Representative arrives to install the instrument you must determine where you want the system placed and the overall layout Consider the factors described in the following paragraphs Space and Accessibility Allow room to interconnect the system components Also arrange for e Comfortable working height e Space for ventilation and access for maintenance and service FC 500 MPL FC 500 MPL w o Second Laser Specifications with HeNe Laser or with Red Solid State Laser Height Instrument 61 0 cm 24 in 61 0 cm 24 in Additional clearance above for servicing and lifting the Data Acquisition card cage above the sensor 45 7 cm 18 in min 45 7 cm 18 in min Total clearance needed 106 7 cm 42 in 106 7 cm 42 in Width Instrument 119 4 cm 47 in 97 8 cm 38 5 in Additional clearance on right for servicing 30 5 cm 12 in 30 5 cm 12 in Additional clearance on left for servicing 30 5 cm 12 in 30 5 cm 12 in Total clearance needed 180 3 cm 71 in 180 3 cm 71 in Depth Instrument 88 9 cm 35 in 88 9 cm 35 in Additional clearance behind instrument for sufficient cooling and room for servicing 30 5 cm 12 in 30 5 cm 12 in Total clearance needed 119 4 cm 47 in 119 4 cm 47 in PN 177579BD 2 1 INSTALLATION SPECIAL REQUIREME
149. ess Voltage pulse signals The signals that the forward scatter side scatter and fluorescence sensors generate They are proportional to the intensity of light the sensor received Well Location The letter and number combination that identifies the well in the plate by row letter and column number position Well Number The number that appears inside the well in the picture of the sample plate on the Plate Wizard GLOSSARY 6 PN 177579BD GLOSSARY Workstation The system component that runs the software that lets you control the instrument It displays sample results and other information PN 177579BD GLOSSARY 7 GLOSSARY GLOSSARY 8 PN 177579BD Symbols BTIM key value 3 9 BYTEORD key value 3 9 CELLS key value 3 9 CYT key value 3 9 DATATYPE key value 3 9 DATE key value 3 9 DFCiToj key value 3 10 ETIM key value 3 9 EXP key value 3 9 FIL key value 3 9 INST key value 3 9 MODE key value 3 9 NEXTDATA key value 3 9 OP key value 3 9 PAR key value 3 9 PnB key value 3 9 PnE key value 3 9 PnN key value 3 9 PnR key value 3 9 PnS key value 3 9 PnV key value 3 9 PROJ key value 3 9 RUNNUMBER key value 3 10 SMNO key value 3 9 PN 177579BD SRC key value 3 9 SYS key value 3 9 TOT key value 3 9 ABSCALFACTOR key value 3 11 ACQUISITIONPROTOCOLOFFSET key value 3 11 BARCODE key value 3 10 BASELINEOF
150. etting of default options for the Listmode Playback Print options publish functions file save naming options are set in this dialog box If you selected Publish Results here and Publish Results to Excel on the Workspace Preferences Publish page this is how the the Excel workbook file is named and saved When using the Replay Plate E feature the workbook file name consists of the Plate ID for the replayed sample plate followed by the File Suffix selected here When not using the Replay Plate feature a maximum of 96 LMD files can be replayed and the workbook file name consists of the first protocol in the listmode playback list followed by the File Suffix selected and the original date and time stamp A new Excel workbook is created for the replayed data The workbook file is saved to the users Results directory The number of worksheets produced depends on the number of protocols used in the worklist and whether you selected Column Format or Experiment Table Format on the Workspace Preferences Publish page File Sorting In the Listcomp setup or Single file mode options listmode files may be sorted using the LMD File Listing option This allows files to be sorted alphabetically or by Time Date modified Files may be Dragged and Dropped Individual files may be selected and moved up or down the list using the arrow buttons The start of each panel is indicated by an arrow B 93 USING MXP SOFTWARE PLOTS MENU A Show File Path checkbo
151. eve the Boolean gating process The Operands are utilized in order to create a combination of regions Gate Logic The shaded areas show the following logical combinations of regions AND Events which are in both regions A AND region B This is written as A AND B F OR Events that are in region A OR in region B This is written as A OR B PN 177579BD B 125 USING MXP SOFTWARE ANALYSIS MENU XOR Events that are EXCLUSIVELY in region A OR in region B but NOT in both This is written as A XORB NOT Events that are NOT in region A This is written as NOT A NOT Events which are NOT in region A AND in region B This is written as NOT A AND B NOT Events which are NOT in region A OR in region B This is written as NOT A ORB B 126 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU PARENTHESES I These are used to define a precise logical statement An open parentheses must be placed after a NOT operator even if the operand is only a single value Example Lymphs AND NOT CD8 Color Blend Mode Introduction MXP software allows the use of combinations of Gates to determine the color of dots in a dot plot The key concept behind this process is similar to color printing and depends upon the use of colored combinations of Gates to define a unique subpopulation Each Gate up to three is given a color and the logical gate combinations give rise to populations with b
152. f a plot changes the associated gate of any region drawn onto this plot also changes to reflect the new gating state Automatic Color Precedence As a new region is drawn the color for that gate is placed at the top of the list in the color precedence dialog box If this option is disabled a color is not associated with a gate B 54 PN 177579BD USING MXP SOFTWARE B FILE MENU Dot Plot Coloring Mode This alternates Color Dot plot mode between Color Blend and Color Precedence modes e Color Blend Mode e Color Precedence Mode default AutoGating Sensitivity Select the sensitivity of the AutoGate region drawn Elliptical Sensitivity Settings are 0 11 0 33 and 1 1 of processed peak Selecting 0 11 creates a larger region whereas selecting 1 1 creates a smaller radius region around the target population Contour Sensitivity Settings are 1 2 3 4 and 5 Level 1 0 11 processed peak at a resolution of 64x64 levels 2 3 and 4 are 0 11 0 33 and 1 1 at a resolution of 128x128 Level 5 is 1 1 processed peak at a resolution of 256x256 Contour Travel Input a maximum value the contour AutoGate region travels from the mean The Minimum value is 25 Maximum is 250 channels Use this option to slightly adjust the AutoGate from sample to sample if slight changes in lysis cause the target population to move slightly This option is not available for elliptical AutoGates AutoGating Failure You can select an error option when
153. f you wish to replace the existing Protocol J Yes e Selecting the No option here allows you to enter a different Protocol File Name Clicking No closes the existing Protocol file and clears the workspace See also Save Protocol As Creating a Protocol for Analysis PN 177579BD B 11 USING MXP SOFTWARE FILE MENU Creating a New Acquisition Protocol 1 D File gt New gt gt New Protocol 2 W Parameters button on the Cytometer Control Acquisition Setup Tab and choose the Parameters to be acquired 3 Create the Plots required for acquisition 4 Ifanalysis is to be performed during acquisition create the required Regions and Gates and also select the required statistics from the Analysis Select Results option 5 O File gt Save Protocol As to save the protocol Creating a Protocol for Analysis 1 File New New Protocol to clear the Workspace of all Regions gates statistical definitions and plots and load the default protocol You can also Drag and Drop a listmode file into the Workspace not into a plot to load the runtime protocol 2 D File Open Listmode File to open a representative listmode data file or Drag and Drop a listmode file into a plot B 12 PN 177579BD USING MXP SOFTWARE B FILE MENU 3 To modify a plot Right mouse click on the plot and Format Plot 4 To create a plot a Choose one of the buttons on the Plots Toolbar or use the Plots Menu to sel
154. factor in the Plate Wizard when defining the Worklist Calculate Results This option calculates the statistics for all files selected on the screen LMD QuickCOMP Enabling LMD QuickCOMP displays orange slider bars on all dual P Lymphs FL3 Log FLS Lin io x fluorescence plots These sliders are used to intuitively adjust the compensation coefficients and update the compensation values in the Cytometer Control Compensation Tab Clicking the arrows adjusts the settings by 0 1 clicking either side of the slider adjusts by 1 0 or drag the slider to place the cells in the required position PN 177579BD B 153 USING MXP SOFTWARE FLOWPAGE MENU Show LMD QuickCOMP Matrix When you use the LMD QuickCOMP slider bars to change compensation settings the new settings are displayed in the LMD QuickCOMP Matrix dialog box You cannot use the LMD QuickCOMP Matrix to change compensation settings Note To view compensation adjustments of Listmode data always view the LMD QuickCOMP Matrix The compensation matrix in the Cytometer Control dialog represents the compensation settings last used in that protocol and may not necessarily be the compensation settings used for any given Listmode file LMD QuickCOMP settings x Compensation Matrix B 21 FLOWPAGE MENU B 154 FlowPAGE Introduction FlowPAGE is the reporting area for Data Plots Statistics and user created te
155. file under a different File Name Use the appropriate procedure depending upon whether you have the standard MXP software or also have the 21 CFR Part 11 option installed Select the appropriate link e Save Listmode File As Procedure With 21 CFR Part 11 Option Installed e Save Listmode File As Procedure Standard MXP Software Installed PN 177579BD PN 177579BD USING MXP SOFTWARE FILE MENU Save Listmode File As Procedure With 21 CFR Part 11 Option Installed Use this procedure if you have the 21 CFR Part 11 option installed on your system 1 D File Save Listmode File As 2 Specify the name of the file a W the desired directory in the Save in list box b J the type of file in the Save as type list box c In the File name text box type the desired File Name Save LMD File Save in 5 color 2 laser autosetup tdf_20040930_115330 fad pe 06_5c 21 as pe comp lmd Sad 10_5c 2l as verify 8 19 3 45 4 Imd e 01_5c 21 as flow set Imd Sad 02_5c 2l as fite comp Imd Bad 03_5c 2l as pe comp Imd Bad 04_5c 2l as ecd comp Imd Bad 05_5c 2l as ape comp imd File name 10_5c 2l as verify 8 19 3 45 4 Imd Save z Cancel Save as type LMD Files Imd fcs Saving As Version 2 Default Comments EMETATEIETEEES Comments for this version adjusted added regions gates 3 You must enter a comment before you Can save a New version e Select an appropriate comment from the list of Default
156. fined by thick black lines and items within the groups are separated by thin dotted lines Acquisition Manager Panel Panel Displays the name of the currently selected panel Protocol AS 2C Flow Set PRO AS HA COARRO TT AS FL2 Comp PRO AS 2C Verify PRO See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST CREATING PANELS Acquisition Manager Protocol Protocol Protocol Displays the name of the currently selected protocol AS 2C Flow Set PRO AS FLACompPRO E EN NS FL CompPRO AS 2C Verify PRO PN 177579BD B 65 USING MXP SOFTWARE VIEW MENU Acquisition Manager Region Settings and Filename Region positions can be carried between different protocols within a panel regions can for example be set on a control sample then carried through all the tests in a panel Note In order for a region position to be carried into a new protocol the source and target region must satisfy all the following criteria e Regions must have the same name e Regions must be of the same type e Regions must be drawn on the same parameters All regions satisfying these criteria are carried from one protocol to another Region Settings There are three options for the region settings This option loads the region positions from the Re E P gion p gion Region Filename current protocol Settings hy This option carries the region positions from those of the pre
157. for this purpose IsoFlow sheath fluid has the following characteristics e Filtered to 0 2 pm e Transparent and nonfluorescent to 488 nm laser light e Low background e Compatible with the characteristics of the sample being measured such as pH osmolality conductivity A 10 L container of sheath fluid is connected to the instrument and sheath fluid is pumped into the internal sheath fluid container as needed The internal sheath container has a working capacity of about 2 L The amount of sheath fluid the container holds beyond the working capacity is for pressurization and level sensing Cleaning Agent When the Cytometer is in the Cleanse mode COULTER CLENZ cleaning agent flushes sample tubing and helps to reduce protein buildup and particles in the instrument Each cleanse cycle uses about 15 mL of cleaning agent The cleaning agent container has a working capacity of about 500 mL That is enough cleaning agent to use the Cleanse mode once a day for 1 month The amount of cleaning agent the container holds beyond the working capacity is for pressurization and level sensing Read the container s label for more information on the cleaning agent PN 177579BD USE AND FUNCTION MATERIAL SAFETY DATA SHEETS MSDS Quality Control Materials The quality control materials available from Beckman Coulter are Flow Check Fluorospheres Flow Set Fluorospheres Cyto Trol Control Cells Immuno Trol Cells Immuno Trol Low Cel
158. g box 4 to display all of the versions of the selected file Note This step is optional Unless you specify a specific version the system opens the most recent version of the file you select B 24 PN 177579BD USING MXP SOFTWARE FILE MENU 5 D M Show Only Signed Files if you only want to view files that have at least one signed version 6 Highlight a file and all of its versions appear below Note Only the signed versions of the file appear if you selected Show Only Signed Files Open LMD File Look in 5 color 2 laser autosetup tdf_20040930_115330 x le 06_5c 21 as pe comp Imd le 02_5c 21 as fite comp lmd be 10_5c 21 as verify 8 19 3 45 4 Imd e7 03_5c 21 as pe comp Imd Bad 04_5c 21 as ecd comp Imd Bad 05_5c 21 as ape comp Imd File name 01_5c 21 as flow setlmd Files of type LMD Files Imd fcs ja Cancel Versions gt gt IV Open Runtime Protocol I Show Archived Files Checked In Versions Version User_ Changed Signed Comments delete 06 Oct 2004 17 04 09 File imported into BCAP I Show Only Signed Files 7 If Yes Click here appears in the Signed field of a version you can O Yes Click here to view the signature information for that version J OK after you review the signatures es Signed Yes Click Here Yes Click Here Filename 4c 1l pro Version 3 Number of Signatures 3 Signature Approved Signed by Mr Anderson Signed on 06 O
159. ge Duration s Sets the maximum Duration of the Acquisition in seconds for each sample When this time limit has been reached acquisition stops However if the maximum number of events is reached before the duration limit acquisition ceases at this point Max Events This is the maximum number of events which may be acquired on the next Acquisition To change this enter the new number required in the input field However if the Duration is reached before the maximum number of events acquisition ceases at this point Drive Space MB Shows how much free data storage space is available on the currently selected storage device in megabytes MB Live Gate This displays all Gates that are available for the Live Gating of data during acquisition Data outside the selected gate is not displayed and is NOT STORED to disk to reduce the size of the listmode files by gating out all unnecessary events PN 177579BD B 165 USING MXP SOFTWARE CYTOMETER MENU B 166 IMPORTANT Improper use of the Live Gate region may generate erroneous but credible results If a Live Gate region is modified during acquisition the data set will be a combination of data from the original and the modifed gate positions If this is not desired then a Restart must be invoked manually To avoid capturing a mixed set of data when using the Live Gate region e Ensure that the Live Gate is correctly positioned on the populations of interest prior to the ac
160. he defaults for the workspace and plot options according to your own preference Some of the options are global so they immediately apply the specific changes required to all plots Other options only apply the changes to the plots created after the option has been selected Use Plots Format Plot to modify the Default Plot options on a per plot basis See also Workspace Preferences Gating Workspace Preferences Publish Workspace Preferences Plot Display Workspace Preferences LMD File Name Workspace Preferences User Info Workspace Preferences Acquisition Options B 44 PN 177579BD USING MXP SOFTWARE FILE MENU Workspace Preferences LMD File Name PN 177579BD This tab allows you to specify a compound listmode file name including components such as the date and the time that the sample was run as well as sample ID fields The File Name displayed in Acquisition Manager is only a provisional file name Some available options for example Run Number and Time can only be defined at the time of acquisition If a printout of the Worklist is required with the final assigned file name it is only correct once acquisition of samples is complete Well Loc is not available The well number is automatically added to the start of the file name for easy identification A file name can have up to 200 alphanumeric characters including the path However the following characters cannot be included forward slash backslash gre
161. he system will not run a sample until the error message is no longer displayed Follow the instructions in the Troubleshooting section for handling this error In order to extend the life of the argon laser when the FC 500 flow cytometer is in idle mode the laser power will be less than 8 mW Once the instrument is taken out of idle mode the laser power will return to 20 mW Thus while operational and out of idle mode the laser power specifications are 20 1 mW Beam Shaping Optics Cross cylindrical lenses 10 mm by 80 mm Argon Laser Beam Spot Size An elliptical spot 10 ym high by 80 ym wide Solid State Laser Beam Spot Size It approximates the beam spot size for the Argon laser Optical Filters For the filters used in the single laser filter block and the dual laser filter block configurations see Figure 5 6 and Figure 5 7 in the Instructions For Use manual e 500 nm long pass LP filter e 530 nm and 620 nm for dual laser configuration short pass SP filter e 488 nm 550 nm 600 nm and 710 nm dichroic long pass DLP filters e 615 nm or 645 nm dichroic short pass DSP filter PN 177579BD PN 177579BD SPECIFICATIONS INSTRUMENT SPECIFICATIONS 525 nm 575 nm 675 nm and 620 nm for single laser configuration band pass BP filters 755 nm long pass LP filter Note The 755 LP filter is functioning effectively as a BP filter for FL5 Sensors The FS sensor and the SS sensor are photodiodes The five FL senso
162. ick the right mouse button on any object on the FlowPAGE to select additional formatting options You can Cut Copy and Paste items that are selected d Cut Copy amp Paste Order k Grouping Properties Align Plot Stats You can change the order of items that are selected db Cut Copy al Paste Grouping r Send to Back Properties Bring Forward Send Backward Align Plot Stats You can group and ungroup items that are selected db Cut Copy l Paste Order gt sro Properties Ungroup B 157 B USING MXP SOFTWARE FLOWPAGE MENU You can edit and change the alignment of text Component Properties gt word Break aa You can change how lines are displayed Component Properties Dashed Dash Dot I Dash Dot Dot You can change the fill properties of items Component Properties that are selected a a hd dizi iv BB NoHatch Downward Diagonal Crosshatch Diagonal Crosshatch Upward Diagonal B 158 PN 177579BD USING MXP SOFTWARE FLOWPAGE MENU You can change the font properties of items that are selected Edit Text Component Properties x Text Line Fil Font Georgia Tp Impact Fp Lucida Console Fp Lucida Sans Unicode Tp Marlett Tp Microsoft Sans Serif T Palatino Linotype Sample Tr Symbol Fp Tah
163. ined from a Protocol other than the current Protocol in memory The Get Cytosettings from LMD file option allows Cytometer Instrument Settings to be obtained from a listmode file The Controls for Acquisition are also found within the Cytometer menu See also Flow Rate Status Bar Cytometer Control The Cytometer Control button show hide dialog box opens the Cytometer Control screen This screen is a See also Cytometer Control Acquisition Setup Tab Cytometer Control Detector Tab Cytometer Control Compensation Tab Flow Rate B 164 PN 177579BD USING MXP SOFTWARE B CYTOMETER MENU Cytometer Control Acquisition Setup Tab This screen is used to set the Acquisition Mode and Acquisition Limits Additionally the Discriminator values for the selected Parameters can be set Duration s x Max Events Acq Setup Detectors Compensation Drive Space MB Acquisition limits Live Gate pame _ Setup Mode Max events o QuickCOMP Mode foo a QuickSET Mode Drive Space MB 1603 4 rs cr Baseline Offset Paseo FLS OFF Live Gate AUX OFF Dots xk gf Ciera Discriminators e z Parameter Button IV QuickCOMP I QuickSET Parameters Red Laser Shutter Qui Parameters M i Red Laser Shutter C Laser oil Laser Enabled See also Cytometer Control Cytometer Control Detector Tab Cytometer Control Compensation Tab Cytometer Control Parameter Setup Flow Rate Status Pa
164. ing your choices Tab Moves cursor over different choices if there are multiple options see Windows manuals for Windows operation via keyboard ait Tab When you have more than one application Window open use Alt Tab to switch between tasks Dialog Box Dialog boxes receive commands or information for example a file name dialog box receives information about a file name Accepts the information you have selected or typed Stops the operation ignoring your choices Description of Reporting Units Unless otherwise stated all parameter units are shown in the US unit format cells pL throughout the manuals GRAPHICS All graphics including screens and printouts are for illustration purposes only and must not be used for any other purpose COMPUTER PROGRAM STATEMENT WARNING This computer program is protected by international copyright laws and unauthorized copying use distribution transfer or sale is a violation of those laws that may result in civil or criminal penalties This computer program may also be subject to additional restrictions contained in a license granted by Beckman Coulter Inc to the authorized user of this computer program or to the authorized owner or other authorized user of the system onto which this computer program is installed Any violation of the license provisions may result in additional civil penalties including an injunction and damages Please refer to the computer program or sy
165. it the dialog box without saving the changes MXP software then updates the display B 104 PN 177579BD PN 177579BD USING MXP SOFTWARE PLOTS MENU Histogram Scaling Clip Channels xi Automatic Maximum Value Labeling StopandSave Font Set Maximum Data Source Histogram Scaling Clip Channels IV Automatic Maximum Value Set Maximum r Cancel Help Clip Channels If Automatic Maximum Value calculation is used then toggling the Clip Channels on ensures that the bottom and top channels are excluded from the calculation of the plots full scale value Automatic Maximum Value This option searches the histogram for the maximum value and then sets the value found to be the full scale for the histogram Set Maximum If the Automatic Maximum Value button is cleared then any value entered here is the full scale for the histogram Histogram Labeling Use the options here to override the default plot display options for the current plot This is useful when capturing images for publication Show Plot Title on Output xi Show Axis Tick Marks Data Source Histogram Scaling Show Axis Labels Labeling Stop and Save Font X and Y Axis Labels IV Show plot title on output IV Show X axis tick marks V Show axis labels IV Show Y axis tick marks V Show Y axis labels X axis label ssTain Pick Y aris label STAN Pick Cancel Help B 105 B USING MXP SOFTWARE PLOTS MENU Co
166. ix Cytometer Control B 170 See also color compensation compensation protocols 3 7 3 8 components See system components computer electrical input requirements 2 2 specifications 3 4 configuration density plot B 107 automatic maximum threshold B 108 density level color B 108 PN 177579BD INDEX logarithmic threshold calculation B 108 maximum density level B 108 smooth B 109 use gate color B 109 use single color range B 109 histogram plot B 101 freeze frame B 102 smooth B 102 use gate color B 102 connections cables location 2 3 power 2 3 signal 2 3 connections tubing location 2 5 pneumatic 2 5 pressure 2 5 vacuum 2 5 waste container 2 5 container waste capacity 2 2 waste connections 2 5 contour plot creating B 106 controls and indicators definition GLOSSARY 2 Cyto Comp cells 1 5 Cyto Trol control cells 1 5 definition GLOSSARY 2 Flow Check fluorospheres 1 5 Flow Set fluorospheres 1 5 fluorospheres 1 5 Immuno Brite fluorospheres 1 5 Immuno Trol cells 1 5 Immuno Trol Low cells 1 5 list 1 5 quality control materials 1 5 QuickCOMP 2 kit 1 5 QuickCOMP 4 kit 1 5 conventions used in the manual xxii copy edit menu B 59 plot data color dot plot B 96 plot image color dot plot B 95 COULTER CLENZ See cleaning agent creating analysis protocol B 12 autogating regions B 135 INDEX 5 INDEX color dot plot B 95 contour plot B 106 density plot B 106 gates B 122
167. kbook used in conjunction with the Tools Publish Results Now menu item The MXP software automatically creates a new workbook when data is published to Excel You cannot overwrite or append an existing workbook The workbook file name is saved to the User s Results directory If you have the 21 CFR Part 11 option when Excel is opened it is under the control of the DaCS software supplied with MXP software DaCS software provides a means of compliance with Regulation 21 CFR Part 11 for data files exported to Microsoft Excel from the MXP software The workbook is under the control of the DaCS software and audit trail information is supplied These Excel worksheets are random password protected and can only be modified when under DaCS software control PN 177579BD USING MXP SOFTWARE B FILE MENU Switch to MS Excel on Publish MS Excel starts and displays the published data When enabled Excel moves to the front of Windows desktop as the active application with each export If left unchecked Excel runs in the background with each subsequent export Publish to MS Excel Format Select the appropriate radio button for the format you want Column Format gt Ifall the tests in the worklist use the same protocol then this format creates one worksheet containing the statistic information of all of the listmode files from the worklist gt If the tests in the worklist use more than one protocol then this format creates a separa
168. ks to be 0 1024 1 024 or 1 10 000 Cytometer Control Translucency Use this slider to fade the Cytometer Controls dialog so it appears translucent ungated SStinfeS FTE rcs Information MEE B rtowpace2 3 Pn 5C 2L AS Verity 8 19 3 45 4 LMD Cytometer Control Acq Setup Detectors Compensation r Acquisition limits Duration s 300 Fs Max events 100000 Drive Space MB 1695 7 r Acquisition mode Live Gate K 7 I Setup Mode I QuickCOMP I QuickSET T Baseline Offset Dots Laser Disabled 1000 G Laser Enabled B 49 B USING MXP SOFTWARE FILE MENU Workspace Preferences User Info When selected this option allows defined information and sample project information to be stored as part of the FCS header in listmode files This can be printed out as required via the Legend plot or with the exported data All of these are optional and can be filled in according to your preference Institute x Institute Street Plot Display Gating Publish Institute Town LMD FileName Acquisition Options User Info Sample Source Institute Facility Project E ell Typ Institute Street Experiment Institute Town Cytometer Serial Number Currently Logged On User Sample Source petae o ooo Project Cell Type Experiment Cytometer Serial Number Currently logged on user AA 1234 Sharlene Cancel Undo See al
169. l In this case the result is reported as 0 00 and should not be considered a valid CV measurement This can happen for all LOG and LIN parameters Note Standard Deviation calculations are only valid between markers that have a Normal or Gaussian distribution CV is not strictly valid for any other shape distribution The derivation of CV can be found in any standard statistic text Median Median is used to indicate the point on the scale of measures where the population is centered The median of a population is the point that divides the distribution of scores in half Numerically half of the scores in a population have values that are equal to or larger than the median and half have values that are equal to or smaller than the median HPCV HP X CV and HP Y CV The Half Peak Coefficient of Variation HPCV is derived by the fixed mathematical relationship between the Standard Deviation SD and the Full Width Half Max FWHM value of a Normal or Gaussian peak The relationship between them is defined as SD FWHM 2 354 This is an approximation to the real value of CV The advantage of calculation HPCV is that the value remains fairly independent of the position of a pair of region markers provided that the markers are set either side of a peak and the marker channel counts are less than half the value of the peak channel count In general the HPCV approximation is usually smaller than the CV calculation See standard statistics texts f
170. l in a cell IQAP Abbreviation for Beckman Coulter s Interlaboratory Quality Assurance Program A service for all worldwide users of Immuno Trol cells and CYTO TROL control cells the IQAP statistically compares your control data with that of other laboratories IVD Abbreviation for For In Vitro Diagnostic Use GLOSSARY 3 GLOSSARY Laser Abbreviation for light amplification by stimulated emission of radiation The standard laser is in the flow cell for analyzing cells Linear amplification See gain Listmode data A list of measurements from each cell Listmode playback tool A tool used 1 to replay 20 bit linear listmode data though new compensation settings 2 for panel playback of listmode files and 3 to replay AutoSetup listmode files to generate a new compensation file LiveGate Listmode Gate used as a live gate in acquisition and for listmode archival LMD Abbreviation for listmode Lock out If you have the 21 CFR Part 11 option you can be manually or automatically locked out of the MXP software You can manually lock the software whenever the instrument will be out of your control but you do not want to log out of the software An automatic lock out occurs when no mouse or keyboard activity has occurred for the time limit set by the System Administrator When the workstation is locked automated activities acquisition listmode playback batch AutoMATOR AutoSetup continue without interruption Log
171. l plots if you have the standard MXP software installed without the 21 CFR Part 11 option In Analysis software this option updates all Plots relevant to one particular sample Before any analysis can be done a listmode file must be read into MXP software Most buttons on the Button Bar are disabled until this has been done To open a listmode file for the first time 1 W File gt Open Listmode File 2 Select the desired listmode file vandv Open LMD File E zix Look in Sy Caspace plus assay B tdf_20030610_113123 vy ek ES 02_caspace pl Well 02 Specimen R_1 Undefined vandv 00000869 2003 06 10 1130 456 LMD 22 04_caspace pl Well 04 Specimen R_1 Undefined vandv 00000870 2003 06 10 1132 457 LMD 06_caspace pl Well 06 Specimen R_1 Undefined vandv 00000871 2003 06 10 1133 458 LMD 08_caspace pl Well 08 Specimen R_1 Undefined vandv 00000872 2003 06 10 1135 459 LMD 10_caspace pl Well 10 Specimen R_1 Undefined vandy 00000873 2003 06 10 1136 460 LMD 12_caspace pl Well 12 Specimen R_1 Undefined vandy 00000874 2003 06 10 1138 461 LMD a the desired directory in the Look in list box The Trays directory has tdf files which contain multiple LMD files 14_caspace pl Well 14 Specimen R_1 Undefined vandv 00000875 2003 06 10 1139 462 LMD 16_caspace pl Well 16 Specimen R_1 Undefined vandv 00000876 2003 06 10 1141 463 LMD 18_caspace pl Well 18 Specimen R_1 Undefined vandv 00000877 2003 06
172. layed Print statistics only Prints all FlowPAGEs If these are not selected the Page icon appears crossed out and a report is not printed from the test Publish Plots or FlowPAGES in PDF format Publish Results Allows the automatic publishing of results to a text file or Microsoft Excel spreadsheet at the completion of each sample depending on the Workspace Preferences selection If publishing to Excel a maximum of 96 files can be replayed in one run The default workbook file name is the current protocol file name with a time and date stamp added The file is saved to the user s Results directory The number of worksheets produced depends on whether you selected Column Format or Experiment Table Format on the Workspace Preferences Publish page B 89 B USING MXP SOFTWARE TOOLS MENU AutoMATOR Status Bar This shows the current file being run and the current state of that file AutoMATOR Status Paused uto start in 15 secs The AutoMATOR Status Bar which displays the function that the test is performing can be removed from the screen Click on the Close button featured on the Status Bar while the AutoMATOR is running This hides the bar from view It automatically reappears again the next time the AutoMATOR is run The Status Bar cannot be brought back onto the screen once closed down without reopening AutoMATOR e Processing This is when the file is being analyzed by the software e Auto Pause When
173. lended colors Consider three gates Gate A set on FL1 positive cells Gate B set on FL2 positive cells Gate C set on FL3 positive cells Any event may be in either none one two or all three of these Gates and therefore combinations of the gates may be used to define eight different subpopulations phenotypes of events as follows Each of these subpopulations may be Default Name Primary A SecondaryB Tertiary C represented by a different color Thus up to Susi FL1 FL2 FL3 eight differently colored subpopulations Group 1 may be visualized on a Color Gated Dot Peh Plot and each group can be given a user Group 4 Group 5 defined name Group amp h 7 Group F Where ve data is inside a Gate and ve data is outside a Gate When in color blend mode ONLY color statistics from the blended colors are displayed in the Legend plot Use to view Region statistics See also Modify Color Blend PN 177579BD B 127 ANALYSIS MENU USING MXP SOFTWARE B 128 Modify Color Blend To use Modify Color Blend 1 W Analysis Modify Color Blend to view the Modify Color Blend dialog box 2 J the Gate you wish to modify 3 Make the desired changes Gate Names Blend Color Enable Name Logic Color Create Gate Modify Color Blend x m Blend Gate 1 Select Population 7 BlendColor IV Enable m Blend Gate 2 Gate Names New Gate 2 7 Blend Color M Enable
174. les AutoMATOR performs batch analysis of stored data using a multi file protocol AutoMATOR allows listmode files to be read and analyzed in a sequential or a predetermined batch order AutoMATOR Setup bx Automator Setup Analysis Files B ilename Modifie 1 C to display the set of files last ar a used If these files are not required el Load Plat aidie highlight them and W Remove Files Reordering Files Eas Load Plate panes Add File or Add Files Remove Files F stn deus ie on Sup Add Blank Soe ne k Panekap P nse E Aatteet Load Queue F Pint FwPAGEs 7 Puish Rests Save Queue Print Restore Analysis Files on Startup Show full pathname Output Options 2 W __AddFie _LoadPlate or Load Queue to select the required file files 3 Navigate to the correct directory and locate the relevant file files PN 177579BD B 85 USING MXP SOFTWARE TOOLS MENU 4 Select the relevant file by highlighting with the mouse If you do not have the 21 CFR Part 11 option you can select more than one LMD file at a time To select more than one file hold down and click the relevant files with the mouse button to deselect click again while is held down 5 O Open The files are listed in the AutoMATOR Setup dialog box Note For the 21 CFR Part 11 option you can repeat steps 4 and 5 as often as needed until all the files you need are listed
175. libration particles FITC amp PE Molecular Probes LinearFlow Intensity Calibration particles APC and IsoFlow sheath fluid Stability Day To Day The mean channel value of the integral signal intensity of Flow Check fluorospheres from the 488 nm laser for FS FL1 FL2 FL3 and FL4 does not vary more than 5 over a period of 7 days when the temperature does not vary more than 5 F from temperature at alignment Refer to the Flow Check fluorospheres package insert for details Stability for FL5 PC7 mean channel value of the integral signal intensity from the 488 nm laser using Flow Check 770 beads does not vary more than 5 from the average of the integral signal intensity mean channel values obtained over a 7 day period when room temperature does not vary more than 5 F from temperature at alignment Refer to the Flow Check 770 fluorospheres package insert for details Stability for FL4 APC mean channel value of the integral signal intensity from the red laser using Flow Check 675 beads does not vary more than 5 from the average of the integral signal intensity mean channel values obtained over a 7 day period when room temperature does not vary more than 5 F from temperature at alignment Refer to the Flow Check 675 fluorospheres package insert for details Within Day The mean channel value of the integral signal intensity of Flow Check fluorospheres from the 488 nm laser for FS FL1 FL2 FL3 FL4 and FL5 does not vary mo
176. light that excited it Fluorescent light FL1 FL2 FL3 FL4 and FL5 sensors PMTs Collect the fluorescent light and generate voltage pulse signals The 1 refers to the first fluorescence sensor 2 the second and so forth Forward scatter FS The laser light scattered at narrow angles to the axis of the laser beam The amount of forward scatter is proportional to the size of the cell that scattered the laser light Forward scatter FS sensor Collects the forward scatter and generates voltage pulse signals Gain The amount of amplification applied to a signal In linear amplification all of a sensor s signals are increased by the same amount Contrast with logarithmic amplification Gating The use of criteria that must be met before an event is included in a histogram GB The abbreviation for gigabyte High voltage Can be adjusted to change the sensitivity of a fluorescent light sensor Histogram A graph showing the relative number and distribution of events HPCYV Half peak coefficient of variation is primarily indicated for bead alignment purposes which is performed using linear scaled data Hydrodynamic focusing A process that focuses the sample stream through the flow cell It ensures that cells move through the laser beam one at a time along the same path Indicators See Controls and Indicators Integral signal A voltage pulse with height and area proportional to the total amount of fluorescent materia
177. lor dot plot B 98 density plot B 109 scaling histogram plot B 105 overlay plot B 110 PRISM plot B 111 PMT definition GLOSSARY 5 FL sensors 3 3 specifications 3 3 PN 177579BD INDEX pneumatic power supply description 1 2 location 2 5 pneumatic tubing connections location 2 5 PNL extension for panel file 3 6 polygonal region creating B 132 interactive information B 134 properties B 132 format tab B 134 See also regions pop up window definition GLOSSARY 5 positives analysis B 136 definition GLOSSARY 5 power cable connections 2 3 electrical input 2 2 power cables connection 2 3 Power Supply definition GLOSSARY 5 description 1 2 dimensions 3 1 electrical input requirements 2 2 See also universal or voltage specific power supply PPP listmode compensation panel extension 3 6 precision reproducibility definition GLOSSARY 5 surface markers performance specifications 3 11 prime Cytometer B 173 region B 136 Prime region definition GLOSSARY 5 print B 43 dots black plot display B 52 FlowPAGE B 43 plot frames plot display B 52 plots B 43 statistics B 43 to PDE B 44 Printer definition GLOSSARY 5 option 1 3 INDEX 17 INDEX PRISM histogram See histogram PRISM plot compatibility B 115 creating B 110 dividers B 113 dual parameter PRISM dividers B 113 B 143 single parameter PRISM divider B 113 B 145 data source B 111 definition GLOSSARY 5 deleting dividers B
178. lorer lets you scroll through the entire list of protocol names Select To position the mouse cursor on an item and then press and release a mouse button to choose that item Sensitivity The ability of the instrument to distinguish very low levels of light scatter and fluorescence from background light or electronic noise Sheath fluid A balanced electrolyte solution Side scatter The amount of laser light scattered at about a 90 angle to the axis of the laser beam The amount of side scatter is proportional to the granularity of the cell that scattered the laser light Side scatter SS sensor Collects the side scatter and generates voltage pulse signals Slider bars They appear on dual fluorescence plots when you enable QuickCOMP mode on the Cytometer Control dialog box These sliders are used to intuitively adjust the compensation coefficients and update the compensation values in the Cytometer Control Compensation Tab SQL Standard query language Standard deviation SD A measure of difference from the mean A measure of precision Toolbar A series of software buttons with related functions USPTO United States Patent and Trademark Office Version number If you have the 21 CFR Part 11 option listmode histogram protocol panel and PDF files receive version numbers The version number is an automatically assigned number incremented by one that the system places at the end of a file as part of the save proc
179. lot This option creates an exact duplicate of the currently selected Plot ctr D may be used as a keyboard shortcut for the same function B 94 PN 177579BD PN 177579BD Color Dot Plot USING MXP SOFTWARE PLOTS MENU The Color Dot Plot option provides two parameter displays of data and allows any combination of the parameters to be selected Each dot in a Color Dot Plot represents one event cell or particle The dot location gives the parameter values for each event Each parameter can have a channel number from 0 to 255 for 256 channel data or 0 to 1023 for 1024 channel data The colors of the dots reflect the Gate or combination of Gates used to create the plot To Create a Color Dot Plot 1 Ow 2 From the dialog box displayed select the required File then select desired Gates Parameters and Regions for display Dot Plot Data Source Dot Plot Events Dot Plot Resolution Dot Plot Labeling Font Stop and Save acquisition only Dotplot Properties 7 Labeling Font Stop and Save Data Source Events Resolution File Gate 5C 2L AS Verify 8 19 3 2 Y x Parameter FL3 Log FL3 Log x I Apply gate to all plots Y Parameter FL4 Log FL4 Log x Regions N Cancel Help 3 When a Plot is displayed click the right mouse button within the plot to display further menu options Copy Plot Image Copy Plot Data Format Plot Copy Plot Image Copy Plot Image Copy Plot
180. lower resolution allows faster redrawing of that image Cancel Help Density Plot Labeling Use the options here to override the default plot display options for the current plot This is useful when capturing images for publication epee T ee Ga T tpat x Show Axis Tick Marks Data Source Density Resolution Show Axis Labels Labeling Font Stop and Save X and Y Axis Labels IV Show X axis tick marks V Show axis labels IV Show Y axis tick marks V Show Y axis labels X axis label ssTaIn Pick Y axis label STAN Pick R Cancel Help B 109 B USING MXP SOFTWARE PLOTS MENU Overlay Histogram Plots The Overlay Plot feature is not available in MXP Software Tomogram Plot The Tomogram Plot feature is not available in MXP Software Surface Plot The Surface Plot feature is not available in MXP Software Prism Plot A Prism Plot is a way of displaying all the subpopulations or phenotype percentages of a multi color immunocytometry analysis protocol The Plot represents the relative percentages of each subpopulation phenotype and the number of phenotypes depends on the number of color parameters being simultaneously analyzed Prism Plots can be customized for example Gate for Prism Scaling and so on To create a Prism plot 1 Og 2 An empty Prism plot window is displayed with the Data Source dialog box overlaying it 3 Make the desired selections Data Source P
181. ls Immuno Brite Fluorospheres Cyto Comp Cell Kit QuickCOMP 2 Kit QuickCOMP 4 Kit Fluorospheres used to check the stability of the optical and fluidic systems Fluorospheres used to standardize light scatter and fluorescence intensity for cell surface marker applications Lyophilized lymphocytes with assay values for specific surface antigens Used to assess monoclonal antibody function and verify proper flow cytometer setup Stabilized erythrocytes and leukocytes with a known quantity of surface antigens Used to verify monoclonal antibody performance as well as verify the process of sample staining lysing and analysis Stabilized erythrocytes and leukocytes with a known quantity of surface antigens Used to verify monoclonal antibody performance as well as verify the process of sample staining lysing and analysis Uniform size fluorospheres with varying fluorescence intensities that are used to monitor instrument linearity Cyto Comp Cells stained with a single color such as CD45 are used to adjust color compensation settings for multicolor analysis using monoclonal antibodies Two single color antibody reagents FITC and PE that can be used to adjust color compensation on a flow cytometer Four single color antibody reagents FITC PE ECD and PC5 that can be used to adjust color compensation on a flow cytometer Additional quality control reagents are available contact your local Beckman Coulter Representative or ac
182. lter block optional 1 3 single laser filter kit optional 1 3 finish button signing on to MXP software B 3 FITC definition GLOSSARY 2 flow cell definition GLOSSARY 2 sensing area 3 1 specifications 3 1 flow cytometry definition GLOSSARY 3 flow cytometry standard definition GLOSSARY 3 flow rate related to pressure 3 2 specifications 3 2 status bar B 174 Flow Check 675 fluorospheres day to day stability 3 12 fluorescence resolution 3 11 log sheets A 1 within day stability 3 12 Flow Check 770 fluorospheres day to day stability 3 12 fluorescence resolution 3 11 log sheets A 1 within day stability 3 12 Flow Check fluorospheres day to day stability 3 12 description 1 5 fluorescence resolution 3 11 log sheets A 1 INDEX 10 within day stability 3 12 See also controls FlowPAGE analysis B 83 analysis results B 83 copy objects B 157 cut objects B 157 data printouts 3 5 definition GLOSSARY 3 deleting objects B 156 edit align bottom B 162 align left B 161 align right B 161 align top B 162 change plot B 160 fill B 157 font B 63 B 157 grow to largest B 160 line B 157 save as B 163 shrink to smallest B 161 space evenly across B 162 space evenly down B 163 text B 159 example B 155 file extension 3 6 insert blank B 74 drag amp drop plot B 78 from file B 75 line B 80 picture B 81 plot B 78 rectangle B 79 statistics table B 76 text file B 82 textbox B 77 introd
183. m Within FlowPAGE B 155 Move a Selected Item Within FlowPAGE B 155 Select a Group of Items Within FlowPAGE B 155 Deleting Objects in FlowPAGE B 156 Resizing Objects in FlowPAGE B 156 Additional FlowPAGE Formatting Options B 157 Edit Text B 159 Change Plot B 160 Grow to Largest B 160 Shrink to Smallest B 161 Align Left B 161 Align Right B 161 Align Top B 162 Align Bottom B 162 Space Evenly Across B 162 Space Evenly Down B 163 Save FlowPAGE As B 163 B 22 CYTOMETER MENU B 164 Cytometer Control B 164 Cytometer Control Acquisition Setup Tab B 165 Duration s B 165 Max Events B 165 Drive Space MB B 165 Live Gate B 165 Setup Mode B 166 QuickCOMP Mode B 167 QuickSET Mode B 167 Baseline Offset B 167 Dots B 168 Discriminators B 168 Parameter Button B 168 Red Laser Shutter B 168 Cytometer Control Parameter Setup B 168 Cytometer Control Detector Tab B 169 Name B 169 Volts B 170 Gain B 170 AUX B 170 Cytometer Control Compensation Tab B 170 Compensation Matrix B 170 Clear All Button B 170 xvi PN 177579BD B 23 B 24 C 1 C 2 PN 177579BD Status Page B 171 Cytometer Log B 171 Get Cytosettings from Protocol B 171 Get Cytosettings from LMD file B 171 Start B 171 Stop B 172 Abort B 172 Abort the Well B 172 Abort the Panel B 172 Abort the Plate B 172 Repeat the Well B 172 Repeat the Panel B 173 Restart B 173 Idle Mode B 173 Prime B 173 Cleanse B
184. me B 103 Delete a Histogram Freeze Frame Overlay B 104 Histogram Scaling B 105 Clip Channels B 105 Automatic Maximum Value B 105 PN 177579BD CONTENTS Set Maximum B 105 Histogram Labeling B 105 Contour Plot B 106 Density Plot B 106 Density Plot Data Source B 107 Density Level Configuration B 107 Density Level Colors B 108 Automatic Maximum Threshold B 108 Maximum Density Level B 108 Logarithmic Threshold Calculation B 108 Smooth Density Plot B 109 Use Gate Color B 109 Use Single Color Range B 109 Density Plot Resolution B 109 Density Plot Labeling B 109 Overlay Histogram Plots B 110 Tomogram Plot B 110 Surface Plot B 110 Prism Plot B 110 Data Source Prism B 111 Prism Scaling Control B 111 What is Prism B 112 How is Prism Created B 112 Prism Dividers B 113 Setting a Single Parameter Prism Divider B 113 Dual Parameter Prism B 113 Editing Prism Dividers B 114 Removing Prism Dividers B 115 MXP Software Prism Compatibility B 115 Legend B 116 Data Source Legend B 117 Legend Options Tab B 117 Show Phenotype Logic Columns B 118 Gated B 118 Total B 118 Number B 118 Absolute Count B 118 Legend FCS Tab B 118 Acquisition Stop and Save B 119 Setting a Stop Counter B 120 Use Stop Condition B 121 Maximum Events B 121 Save Histogram Data in FCS Format B 121 Format Plot B 121 B 20 ANALYSIS MENU B 122 Create Modify Gates B 122 Gate and Logical Expression B 122 Ne
185. ministrator s Guide 2 the CD ROM containing the System Administrator s Guide should be protected from access by non administrator personnel ABOUT THIS MANUAL Your Cytomics FC 500 MPL Reference manual provides instrument specifications and information on installation safety and software features This information is organized as follows Chapter 1 USE AND FUNCTION Contains a short description of the major instrument components and options and the reagents and quality control materials used Chapter 2 INSTALLATION Contains instrument requirements and diagrams of the interunit cable connections Chapter 3 SPECIFICATIONS Details the instrument and performance specifications Appendices The appendices provide reference material on the following topics gt LOG SHEETS gt USING MXP SOFTWARE gt 21 CFR PART 11 OPTION COMPLIANCE INFORMATION PN 177579BD xxi INTRODUCTION CONVENTIONS GLOSSARY The glossary has definitions for many of the words and terms used in flow cytometry INDEX Use the Index to easily locate specific information in this manual CONVENTIONS This manual uses the following conventions xxii Throughout this manual your FC 500 MPL is also referred to as the system or instrument Bold font indicates a software option such as Cytometer Italics font indicates screen text displayed on the instrument such as Preparing Samples Courier font indicates text you have to type using th
186. n Acquisition Manager ensure that you select Use Plots and Gates from Previous Test in the Panel Wizard for the panel 1 Read ina file or acquire data to be assigned as an overlay for example a negative control 2 Highlight the chosen single parameter Histogram to be used as an overlay plot click on the title bar 3 Click the right mouse button on the plot and O E Format Plt then C the Histogram tab See also Histogram Formatting 4 Check a Freeze Frame box to assign the current Histogram The top button creates a dotted overlay and the bottom button creates a dashed Histogram 5 The current Histogram is assigned to whichever overlay button is selected and its name appears in the edit box adjacent to the button this legend may be modified if desired 6 O or press to accept any changes 7 D or press to exit the dialog box without saving the changes Now you can open another file or acquire a positive sample The new data is displayed as the base histogram and the previous sample is seen as an overlay B 103 B USING MXP SOFTWARE PLOTS MENU Delete a Histogram Freeze Frame Overlay 1 Highlight the plot containing the unwanted overlay click on the title bar 2 Click the right mouse button on the plot and O E Format Plot then W the Histogram tab 3 Clear the appropriate Overlay Histogram checkbox to delete an overlay 4 J or press to accept any changes 5 or press to ex
187. n be used for each Sample ID However if you open this plate definition file tdf again in the Plate Wizard the edited values for Sample IDs 2 3 and 4 return to their original values By default the Sample ID fields in the Acquisition Manager correspond to the following fields in the Plate Wizard e Sample ID 1 corresponds to the Sample ID entered in the Plate Wizard e Sample ID 2 corresponds to Well Type e Sample ID 3 corresponds to the Replicate number e Sample ID 4 corresponds to Dilution value entered in the Plate Wizard only applies to calibrators and controls See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST PN 177579BD USING MXP SOFTWARE B VIEW MENU Acquisition Manager Cal Factor Cal Factor Displays the user defined Cal Factor entered in the Plate Wizard when using absolute count calibration particles with an assay count Cal Factor See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST Absolute Count Calibration Acquisition Manager LMD File Name LMD File Name The File Name displayed in this column is only a provisional file name Some available options for example Run Number and Time can only be defined at the time of acquisition If these elements are included in the file name and a printout of the Worklist with the final assigned file name is re
188. n there should be no effect at all by using Baseline Offset on Statistics data from that region This function allows log data in very low level signals to be moved up from low numbered channels by adding a randomized Gaussian positive offset This allows easier comparison of data from other Cytometers Baseline Offset includes the following three options e Auto This option displays the listmode data according to a keyword in the FCS file header If the Baseline Offset keyword value is ON then data is automatically displayed with Baseline Offset switched on If it is OFF then data is displayed with no Baseline Offset e On This reads a file and modifies the displayed data only by adding a randomized Gaussian offset to the low level signals Data on disk is not modified and is left as it was when acquired from the Cytometer e Off Reads a file exactly as it is without modification during display Note Changing Baseline Offset settings on the Cytometer Control dialog does not affect the currently loaded listmode file Only changes made on the Workspace Preferences dialog affect the currently loaded listmode file If the desired Baseline Offset option is already selected in Workspace Preferences select another Baseline Offset option apply and then select the desired Baseline Offset option again Default Overlay Mode This feature is not available in MXP software Default 3D Plot Mode This feature is not available in MXP software
189. n to delete and O Bere Use ma to select multiple regions to delete Use Cti A to select all of the regions 3 D or press to delete the Regions C or press to exit the dialog box without deleting any selected Regions When you exit the View Modify Regions dialog box MXP software updates all the Windows If a deleted Region was also specified in any statistics or Gates then MXP software updates to reflect these changes Re Calculate All AutoGates Use this function to re calculate multiple autogates in a protocol rather than re calculating them individually B 142 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Create Prism Divider Dual Prism Divider The Analysis gt gt Create Prism Divider gt gt Dual Prism Divider menu option allows you simultaneously set two Prism dividers in a quadrant fashion on a dual parameter plot Any displayed Prism Plot is automatically updated with the new Prism statistics If the number of Prism dividers is changed during the divider creation the corresponding Prism plot also reflects this by changing its format according to the number of active Prism dividers Note MXP Cytometer software only allows one Prism divider per parameter and the dividers are GLOBAL across the application As a consequence modifying the Prism divider of any parameter on any plot automatically updates the divider of that parameter if it is displayed on any other plot This includes both dual and single
190. ncluding flow rate data rate number of parameters collected gated events data plots FlowPAGE reports and printouts generated As an example a maximum cycle time is 45 seconds well or tube when aspirating 100 pL of Flow Check fluorospheres using the QC 1L Flow Check standard protocol Changing these variables such as decreasing the number of events collected increasing sample concentration or increasing the flow rate can increase your acquisition throughput while increasing the number of events collected or running at a lower data rate can decrease your acquisition throughput Precision for Surface Markers See reagent package insert for precision specifications of other surface markers Resolution PN 177579BD Forward Scatter The HPCV of the integral signal intensity values using Flow Check fluorospheres is lt 2 Fluorescence The HPCY of the integral signal intensity values using Flow Check fluorospheres is lt 2 for FL1 FL4 from the Argon laser The HPCV of the integral signal intensity values using the Flow Check 675 fluorospheres is lt 2 5 for FL4 from the red laser The HPCV of the integral signal intensity values using Flow Check 770 fluorospheres is lt 4 for FL5 3 SPECIFICATIONS PERFORMANCE SPECIFICATIONS 3 12 Sensitivity Scatter Detects 0 5 pm 40 pm diameter particles Fluorescence 600 FITC 300 PE and 600 APC molecules of equivalent fluorochrome when measured with Spherotech Rainbow Ca
191. nt Statistics er PM Save Protocol Print FlowPAGES Note This step only applies if you are IF Publish I Pin FlowPAGES To POF using one of the Analysis Add Ins to T Print Plots To PDF export data files to Excel during acquisition PN 177579BD B 17 USING MXP SOFTWARE FILE MENU 6 C the Publish tab 7 C Iw Publish Data to MS Excel 8 O Experiment Table Format o box Iv Switch to MS Excel on publish Optional If this option is chosen the system displays the Excel spreadsheet after each sample is run during acquisition LMD FileName Acquisition Options Userinfo Plot Display Gating Publish Export Bitmap Size Small 400x400 Medium 600x600 C Large 800x800 File References in Data F Full Path amp Name Name Only I Publish Data to Text File IV Publish Data to MS Excel IZ Switch to MS Excel on publish Publish to MS Excel Format 10 Only files using the same protocol should be sent to Excel when using either of the Analysis Add Ins B 18 For creation of Excel files during data acquisition assign the same protocol to all wells of a sample plate This is done in the MXP Plate Wizard For LMD Playback Tool you must select the same protocol to analyze all of the files For AutoMATOR or an open LMD file on your desktop only one protocol applies automatically PN 177579BD PN 177579BD USING MXP SOFTWARE FILE MENU
192. ntour Plot The Contour Plot feature is not available in MXP Software Density Plot The Density Plot option provides two parameter data display any combination of parameters can be selected Each colored area in a Density Plot represents an ISO count of the number of events cells or particles The color shows the number of events at any X and Y parameter value To create a Density Plot 2 From the Data Source dialog box x displayed select the required File then Eo EER E select desired Gates Parameters and DataSource Density Resolution Regions for display File Gate Density Plot Data Source Density Level Configuration Lal Malia i FS Lin FS Li 7 SS Lin SS Li z Density Plot Resolution uses in SS Lin Density Plot Labeling J Apply gate to all plots ss Stop and Save acquisition only Cabal Help 3 When a Plot is displayed click the Copy Plot Image right mouse button to display further Copy Plat Data menu options Format Copy Plot Image Copy Plot Data Format Plot B 106 PN 177579BD USING MXP SOFTWARE B PLOTS MENU Density Plot Data Source Choose the X and Y parameters for the current plot Gating equation and regions to be displayed can be selected File x Gate Selection Labeling Font Stop and Save X and Y Parameter DataSource Densty Resoltion Apply Gate to All Plots Fie ee Regions 5C 2L AS Verity 8419 34 gt x x Parameter Y Parameter
193. o Brite fluorospheres description 1 5 See also controls Immuno Trol cells description 1 5 See controls Immuno Trol Low cells description 1 5 See controls import settings from CMP file B 42 indicators definition GLOSSARY 3 input electrical special requirements 2 2 INDEX 12 insert FlowPAGE blank page B 74 drag amp drop plot B 78 from file B 75 line B 80 picture B 81 plot B 78 rectangle B 79 statistics table B 76 text file B 82 textbox B 77 menu B 74 overlay histogram B 83 overlay plot annotation B 83 overlay plot auto text B 83 installation category 3 1 MXP software 2 6 institute address B 50 user info B 50 instrument accessibility 2 1 delivery inspection 2 1 dimensions 3 1 electrical input requirements 2 2 installation special requirements 2 1 layout 2 1 location 2 1 monitor linearity 1 5 performance specifications 3 11 software specifications 3 5 space needed 2 1 unpacking 2 1 view l 1 instrument settings definition GLOSSARY 2 integral signal definition GLOSSARY 3 See also signals intensity calibration particles used in fluorescence performance specifications 3 12 introduction to your manuals xxi IQAP definition GLOSSARY 3 IsoFlow sheath fluid See sheath fluid PN 177579BD IVD definition GLOSSARY 3 K keyboard cable connection 2 3 Kit dual laser filter 1 3 file extension 3 6 filter 1 3 single laser filter 1 3 L labeling color dot plot
194. o not need the negative control pro or Flow Set pro protocols These ppp panels allow you to replay the associated LMD files to create cmp files Runtime panels may be selected as these contain Flow Set fluorospheres protocols which have no use in this dialog this protocol is removed from the displayed panel and a warning is displayed The dialog is resizable by dragging the corners to the required size The settings file name defaults to the name of the panel but with a new cmp extension The Auto save checkbox allows settings to be automatically written to the settings file that has the name of the panel selected Panels can be created within this dialog but cannot be used for acquisition These listmode panels are saved with a ppp extension The Parameter field indicates the Compensation parameter within the protocol The Status field allows a pause or stop between each file playback Note Listmode Compensation must be performed using the runtime protocol or a protocol with equivalent parameters as the runtime protocol PN 177579BD PN 177579BD USING MXP SOFTWARE B TOOLS MENU Options Tab Ol RAPA XK gt n u i S Single File Mode List Comp Setup Options Output _ Preferences T Minimise when running I Print Stats I Show File Paths JP Print FlowPAGEs I Show Region Filename I Publish Results File Suffix I Publish FlowPAGE as PDF modified JT Publish Plots as PDF This tab allows s
195. oma AaBbCcD zl I Underline J Strike Dut Text Color N 1 W Textbox in the FlowPAGE that you wish to edit The edit handles appear 2 Right mouse click on the textbox and C Properties db Cut Copy l Paste Order Grouping gt 3 Type in the new text 4 x See Edit Font FlowPAGE PN 177579BD Component Properties xj Test Line Fil Font Position and Size Text Edit FlowPAGE tex IV Multiple Lines I Word Break Horizontal Alignment Vertical Alignment C Left Top Center Center Right Bottom B 159 B USING MXP SOFTWARE FLOWPAGE MENU Change Plot Use Change Plot to change which Plot is displayed from those available on the MXP desktop 1 On the FlowPAGE J the Plot you want to replace The edit handles appear Select Plot to Use xj Lymphs FL3 Log FL5 Log Lymphs FL2 Log FL3 Log Lymphs FL1 Log FL5 Log Ungated SS Lin FS Lin P Ungated Legend Lymphs FL4 Log FL5 Log Lymphs FL3 Log FL4 Log Lymphs FL5 Log FL4 Log Lymphs FL2 Log FL4 Log Lymphs FL1 Log FL4 Log Lymphs FL1 Log FL3 Log 5000 Lymphs FL1 Log FL2 Log LA funmnhs FL an xl 2 W FlowPAGE gt gt Change Plot 3 Select the plot you want to use Cancel Help 4 Oma Grow to Largest 1 Select the items to be changed See Select a Group of Items Within FlowPAGE 2 al All i
196. on Setup Tab The incoming data is not saved Acquisition During data acquisition the plots are updated in real time When one plot is displayed with statistics underneath the statistics are also updated in real time Unlimited one or two parameter plots are available for any given sample One parameter plots have 1 024 channel resolution Two parameter plots have up to 512 x 512 channel resolution Regions Up to 32 gates and 256 analysis regions are available for gating analysis and autogating per protocol Up to 8 of those regions can be used to create any one gate The following types of regions are available for gating and analysis Linear Rectangular Quadrant Polygonal Elliptical Contour 3 5 3 SPECIFICATIONS SOFTWARE SPECIFICATIONS 3 6 Listmode Analysis The instrument can store up to 16 parameters including AUX TIME and RATIO as listmode data The amount of available random access memory RAM in the computer determines the maximum size of a listmode file MXP Default File Extensions MXP has a variety of standard file extensions Many of these file extensions are not necessary for MXP itself but are required for compatibility with other programs Panel Protocol and Worklist files are written as pure ASCII text files and can be read and edited by any ASCII text editor such as Windows Notepad or the MS DOS Edit command The advantage of using ASCII text to describe the set up information means that
197. on Linked Marker B 137 View Modify Regions B 138 Regions B 138 Region Points B 139 Resizing a Region B 139 Moving a Region B 139 Editing and Moving a Region Name B 140 Editing a Quadrant Region Name B 141 Deleting Regions B 141 From Within a Plot B 141 Using the Delete Button in the View Modify Regions Dialog Box B 142 Re Calculate All AutoGates B 142 Create Prism Divider B 143 Dual Prism Divider B 143 Setting a New Dual Prism Divider B 143 Editing a Dual Prism Divider B 144 Single Prism Divider B 145 Setting a New Single Prism Divider B 145 Editing a Single Prism Divider B 146 View Modify Prism B 147 Select Results B 148 Statistic Type B 149 Current File Name B 149 Mean Calculation Method B 149 FCS Information B 149 Number B 149 Total B 149 Gated B 149 Percentile B 149 Mean B 149 CV B 150 Median B 150 HPCY B 150 Mode B 150 Cells yL B 150 Min and Max B 150 Advanced Statistics Configuration B 151 Differences Between Log Log and Log Lin Statistical Calculations B 151 Log Log B 151 Log Lin B 151 Automatic Statistic Percentiles B 151 Absolute Count Calibration B 152 Setting a CAL Factor for Absolute Counts B 152 Advanced Button B 153 Setting a CAL Region B 153 CONTENTS XV CONTENTS Calculate Results B 153 LMD QuickCOMP B 153 Show LMD QuickCOMP Matrix B 154 B 21 FLOWPAGE MENU B 154 FlowPAGE Introduction B 154 Editing FlowPAGEs B 155 Select a Single Ite
198. ons with Voltage Specific Power Supply 2 5 Waste and Pneumatic Tubing Connections with Universal Power Supply 2 6 xix CONTENTS XX PN 177579BD INTRODUCTION D This introductory section contains the following topics e USING YOUR Cytomics FC 500 MPL MANUALS e ABOUT THIS MANUAL e CONVENTIONS and e GRAPHICS USING YOUR Cytomics FC 500 MPL MANUALS The manuals listed below are available as PDF files in the FC 500 MPL MXP software Printed versions of these manuals are also available by order Use the Reference manual for instrument specifications and information on installation and system options Use the Instructions For Use manual for the day to day running of your instrument Go through the detailed step by step procedures of startup quality control QC running samples analyzing data printing reports reviewing QC data and shutdown It contains safety and troubleshooting information error messages as well as in depth information on the principles of flow cytometry information about what your instrument does and the methods it uses Use the Special Procedures manual to clean replace or adjust a component of the instrument Use the System Administrators Guide to add and edit user profiles including their User IDs and privileges If you have the 21 CFR Part 11 optional software as part of your laboratory s responsibility for compliance 1 only the System Administrator should have access to and use the System Ad
199. operties B 134 quadrant region B 135 properties B 135 PN 177579BD INDEX re name B 141 rectangular region B 134 interactive information B 135 properties B 134 setting a CAL region B 153 specifications 3 5 toolbar B 6 types available for gating and analysis 3 5 view modify B 138 delete region B 141 move region B 139 region name B 140 region points B 139 resize region B 139 select region B 138 report See FlowPAGE reproducibility precision definition GLOSSARY 5 surface markers performance specifications 3 11 requirements installation instrument accessibility 2 1 space needed 2 1 requirements sample 3 1 resolution color dot plot B 98 density plot B 109 performance specifications 3 11 resource explorer B 70 restart Cytometer B 173 Result Excel worksheet C 5 results advanced statistics configuration automatic statistic percentiles B 151 differences between log log and log lin statistical calculations B 151 log lin B 151 log log B 151 select B 148 advanced statistics configuration B 151 current file name B 149 FCS information B 149 statistic type B 149 romlock definition GLOSSARY 6 room temperature ambient operating 2 2 INDEX 19 INDEX special requirements 2 2 specifications 2 2 run number LMD file name B 46 runtime protocol See protocols S sample ID 1 2 3 4 LMD file name B 46 info display in acquisition manager B 68 measurable cell sizes 3 1 optim
200. or further explanation of HPCV and CV differences Mode The mode channel is the channel with the largest number of counts between region markers Gate colors can be customized using the drop down color box and selecting the required color Cells pL This displays the absolute count of a population based on the calibration factor CAL Factor and the number of particles within a user defined CAL region Min and Max Displays linear region boundaries B 150 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Advanced Statistics Configuration Selecting the Advanced button offers further Advanced Statistics CONANA E options to select the way that the mean is r Calculation Preference calculated a lies OK i z 5 Elite Lysys Cancel Log Log Help Log Lin m Automatic Statistic Percentiles Automatic Statistic Percentiles X Axis Y Axis joo Boo Differences Between Log Log and Log Lin Statistical Calculations For most practical applications users may never notice the difference between the two In general Log Log mean appears to have its value skewed further to the right than a Log Lin mean calculation on the same data This skew is more pronounced when the distribution of data extends greater than half a decade from the mean value however calculated Log Log Selecting Log Log calculates the arithmetic mean values of Log data using conversion of each individual channel before a mean
201. ort or uncheck the Setup Mode checkbox and then perform the acquisition PN 177579BD USING MXP SOFTWARE B CYTOMETER MENU QuickCOMP Mode Enabling QuickCOMP on the Cytometer Control Acquisition Setup Tab dialog box displays red slider bars on all dual fluorescence plots P Lymphs FL3 Log FES eee m Es These sliders are used to intuitively adjust the compensation coefficients and update the compensation values in the Cytometer Control Compensation Tab Clicking the arrows adjusts the settings by 0 1 clicking either side of the slider adjusts by 1 0 or drag the slider to place the cells in the required position When QuickCOMP is enabled QuickSET mode is not available Note Listmode Compensation must be performed using the runtime protocol or a protocol with equivalent parameters as the runtime protocol QuickSET Mode Enabling QuickSET on the Cytometer Control Acquisition Setup Tab dialog box a displays slider bars on all plots These sliders are used to adjust voltages at the data plots and update the high voltages in the Cytometer Control Detector Tab Clicking the arrows adjusts the voltage clicking either side of the slider adjusts the voltage by a higher increment Or drag the slider to place the cells in the required position When QuickCOMP is enabled QuickSET mode is not available Baseline Offset Enable this checkbox to switch on Baseline Offset Further information
202. orts the method of calculating the mean as part of a Published Excel spreadsheet FCS Information These options are either generated automatically by MXP software or obtained from sample information entered by you Desired items can be selected and exported during the publishing of an Excel Spreadsheet The selected FCS information is only published and does not print with the plots within MXP software Number Total number of events in a Region Total Total number of events in region as a percentage of total events in the file Gated Total number of events in region as a percentage of events in a gated display Percentile This is the channel which divides the events in a region by the percentile value The default percentile is 50 the median but this can be set to request any desired value Mean The Mean channel is the arithmetic mean channel defined as the sum of the region channel numbers times counts divided by the integral of the region The Mean or the average is found when all selected events are added together and then divide by the total number of events The Advanced button allows for the calculation of a Geometric mean PN 177579BD B 149 USING MXP SOFTWARE ANALYSIS MENU CV The coefficient of variation or CV is computed by dividing the standard deviation by the mean of the data between the markers An accurate CV cannot be computed when there are insufficient events to make the value meaningfu
203. ou want to view PN 177579BD B 34 PN 177579BD USING MXP SOFTWARE FILE MENU 3 to display all of the versions of the selected file Note This step is optional Unless you specify a specific version the system opens the most recent version of the file you select 4 I Show Only Signed Files if you only want to view files that have at least one signed version 5 Highlight a file and all of its versions appear below Note Only the signed versions of the file appear if you selected Show Only Signed Files Open Protocol Look in AcquisitionProtocol x 2 AS 2C Settings PRO BAS AC IL BAS FLIC HH as 3c 11 settings pro He as 3c 11 flow set pro a 5c 2 pro SSS 1 File name eteo o Open Files of type Protocol Files pro Cancel Checked In Versions Version User_ Changed Signed Commens delete 06 Oct 2004 17 14 48 Yes Click Here updated flow pages delete 06 Oct 2004 17 14 39 Yes Click Here added plots delete 06 Oct 2004 17 13 59 File imported into BCAP I Show Archived Files I Show Only Signed Files 6 If Yes Click here appears in the Signed field of a version you can W Yes Click here to view the signature information for that version D OK after you review the signatures Signed Yes Click Here Yes Click Here Filename 4c 1l pro Version 3 Number of Signatures 3 Signature Aporoved Signed by Mr Anderson Signed on 06 Oct
204. ower Supply Cytometer Flow restrictor box T Pressure Flow cell fluid Vacuum 2 i el evel Waste La min ISOFLOW SHEATH FLUID WASTE Waste Waste vent 20L Waste level 10 L sheath fluid waste container container 7579003B PN 177579BD 2 5 INSTALLATION INSTALLING MXP SOFTWARE Figure 2 4 Waste and Pneumatic Tubing Connections with Universal Power Supply Cytometer back cover Cytometer TANGER __ _ Flow restrictor box ce Sheath d fluid level Flow cell a waste Waste ee k Sheath T fluid Waste vent J Waste level 10 L sheath fluid waste container container 7579047B 2 4 INSTALLING MXP SOFTWARE To install the MXP software see the System Administrators Guide 2 6 PN 177579BD SPECIFICATIONS 3 1 SAMPLE REQUIREMENTS See SAMPLE REQUIREMENTS in the RUNNING SAMPLES chapter in the Instructions For Use manual for details 3 2 INSTRUMENT SPECIFICATIONS Dimensions Component Height Width Depth Weight Computer 15 75 cm 53 34 cm 42 55 cm 18 14 kg 6 2 in 21 in 16 75 in 40 1b Cytometer 61 0 cm 97 8 cm 105 4 cm 93 9 kg and MPL 24 in 38 5 in 41 5 in 207 lb only for units with Universal Power Supply Cytometer 61 0 cm 97 8 cm 88 9 cm 84 8 kg and MPL 24 in 38 5 in 35 in 187 lb Monitor 42 7 cm 40 5 cm 43 8 cm 22 kg typical 16 81 in
205. panels are not opened closed removed and or replaced with care e Improper tools are used for troubleshooting To avoid injury e Keep doors covers and panels closed and secured in place while the instrument is in use e Take full advantage of the safety features of the instrument Do not defeat safety interlocks and sensors e Acknowledge and act upon instrument alarms and error messages e Keep away from moving parts Report any broken parts to your Beckman Coulter Representative Open remove and close replace doors covers and panels with care Use the proper tools when troubleshooting CAUTION System integrity might be compromised and operational failures might occur if e This equipment is used ina manner other than specified Operate the instrument as instructed in the Product Manuals e You introduce software that is not authorized by Beckman Coulter into your computer Only operate your system s computer with software authorized by Beckman Coulter e You install software that is not an original copyrighted version Only use software that is an original copyrighted version to prevent virus contamination IMPORTANT If you purchased this product from anyone other than Beckman Coulter or an authorized Beckman Coulter distributor and if it is not presently under a Beckman Coulter service maintenance agreement Beckman Coulter cannot guarantee that the product is fitted with the most current mandatory engineering revisions or that yo
206. parameter plots See also Setting a New Dual Prism Divider Editing a Dual Prism Divider Setting a New Dual Prism Divider 1 Highlight the chosen dual parameter plot current window 2 W Analysis gt Create Prism Divider gt Dual Prism Divider 3 A Dual Prism cursor is displayed in the current window 4 Position the cursor anywhere on the window click to show the prism divider lines If is pressed at this point the operation is abandoned 9 Move cursor to position crosshair then click to set position PN 177579BD B 143 USING MXP SOFTWARE ANALYSIS MENU Editing a Dual Prism Divider Editing an existing Dual Parameter plot with a set of dual Prism dividers displayed essentially consists of creating a new set of dual Prism dividers When the second set of dual Prism dividers is defined the first set is deleted 1 Highlight the required dual parameter plot 2 Mouse over the Prism divider to get the tor cursor 3 hold to set the new Dual Prism crosshair When the new dividers are set they replace the previously set dividers Any associated Prism plot is updated with the new Prism statistics Any associated Prism plot changes its format and has its statistics updated to reflect a smaller number of Prism dividers and a smaller number of displayed phenotypes 4 To delete Prism divider on the divider to make it active and press Delete The Prism divider for the associated parame
207. peat the panel B 173 repeat the tube B 173 about this manual xxi absolute calibration particles in acquisition manager B 69 setting a Cal factor B 152 setting a CAL region B 153 absolute counts calibration B 152 setting a Cal factor B 152 calibration advanced button B 153 accessibility instrument installation requirements 2 1 accuracy definition GLOSSARY 1 acquisition stop and save B 119 stop counter maximum events B 121 minimum count B 136 save histogram data in FCS format B 121 use stop condition B 121 acquisition manager acquisition options B 49 Cal factor B 69 instrument file name B 67 instrument settings B 67 introduction B 64 LMD filename B 69 panel B 65 parameter names B 67 protocol B 65 region sources B 66 reorder worklist columns B 65 sample info B 68 toolbar B 7 well info B 68 worklist columns available B 65 See also worklist acquisition options acquisition manager B 49 cytometer control translucency B 49 default log scaling B 49 output options B 49 workspace preferences B 48 PN 177579BD acquisition setup Cytometer Control B 165 baseline offset B 167 discriminator B 168 dots B 168 drive space B 165 duration B 165 live gate B 165 max events B 165 red laser button B 168 setup mode B 166 Action log sheet A 1 activities automated See automated activities Add In See MPL DaCS Analysis Add In and MPL Analysis Add In admin button signing on to MXP software
208. quired they only print correctly once acquisition of samples is complete The well number is always placed at the start of each listmode file name LMD Filename 01 _00000023 017 LMD 03_00000025 019 LMD 04_00000026 020 LMD See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST PN 177579BD B 69 USING MXP SOFTWARE VIEW MENU Resource Explorer The Resource Explorer occupies a definable area within the Workspace It allows you to browse files and can be used to Drag and Drop items into MXP software Tabs for Listmode files and Protocols allow files to be dropped into the Workspace Listmode file tab displays the Listmode directories for the logged in user s Trays directory Tabs for Panels and Histograms allow you to view all the panels and histograms for the selected directory Resource Explorer With 21 CFR Part 11 Option 01_5c 2l as flow set Imd 02_5Sc 2l as Fite comp md 03_5c 2 as pe comp md 04_5c 21 as ecd comp Imd 05_5Sc 2 as ape comp Imd 06_5c 2l as pc comp md 07_5Sc 2l as verify 8 19 3 45 4 5c 11 16 56 3 19 45 00000114 1 B 5C 2L Color Worklist tdf HH 07_5c 21 as verify 8 19 3 45 4 PRO v 2 H fg Plots bf Regions J Gates y LMD Files y 07_5c 2l as verify 8 19 3 45 4 Imd v 5 Resource Explorer Standard MXP Software A user Acquistion File Type Date Time E user Analysis
209. quisition of data Do not make changes to the Live Gate region or assignment during acquisition e Make changes to Live Gate regions only after activating the Setup Mode option in the Cytometer Control screen resulting in a refresh of the data If the MXP software is not in Setup Mode when changes are made to a Live Gate region a manual Restart is required Review all data plots and results prior to reporting results Setup Mode Enable this checkbox to allow the continuous acquisition of data from the Cytometer In this mode Stops are disabled and listmode files are not stored Setup Mode allows continuous acquisition of data from the Cytometer giving continuous real time feedback of instrument performance on the displayed plots Any Stop amp Save values set are ignored in this mode Display of data continues until Abort is selected to end data acquisition or the Setup Mode box is unchecked to begin normal data acquisition In Setup Mode the plots are updated live time with only the most recently acquired events displayed The incoming data is not saved During Setup Mode older events are cleared from the screen and from memory allowing the Cytometer settings to be adjusted in real time without reaching a set stop condition Data is displayed on a first in first out basis The number of events to be displayed can be changed by entering the required value into the Dots input box Once you have finished using Setup Mode click Ab
210. r Region Properties Region Name eionrroneries Lower Limit and Upper Limit Region Name Neus Prime Target Region for Automatic Setup Lower limit 0 1024 Upper limit 0 87881 Automatic Region Name Positioning J Prime J Target region for automatic setup Positives Analysis IV Automatic region name positioning Interactive Linear Region Editing Positives Analysis I Active Percent Positive p Minimum Count a T Active Count fp Cancel Help Lower Limit and Upper Limit Set the region to defined limits by entering upper and lower limits for the region These limits can be viewed in the statistics box for each plot by selecting Analysis Select Results and select Min Max Target Region for Automatic Setup Specify the region as the Flow Set fluorospheres target region for AutoSetup when multiple regions are present on a FS plot Positives Analysis This option moves the region to the selected percent result Specify a positive percentage value from 0 1 to 99 9 If the positive analysis option is not selected the percentile is unavailable Use this option to automatically set the region position for a negative control sample Interactive Linear Region Editing When a Region s position is changed any Gates that use the Region are automatically updated and each plot is redrawn to reflect the changes in the Gate When the Region is deleted any Gate which used the Region is also automatically deleted When a
211. r worksheets DaCS software provides the controls to help you bring your laboratory into 21 CFR Part 11 compliance for data files exported to Microsoft Excel from the MXP software When you log into MXP software e A DaCS progress bar screen briefly appears as the DaCS software is started e No additional actions are needed after you log into the MXP software to be automatically logged into the DaCS software when data is exported to MS Excel PN 177579BD 21 CFR PART 11 OPTION COMPLIANCE INFORMATION DaCS SOFTWARE When you log out of MXP software e All open Excel workbooks are saved and closed e You are automatically logged out of the DaCS software The MXP DaCs Interface Workbook is opened when data is exported to MS Excel from the MXP software A_SYSTEM and an _AUDIT worksheet and tab always appear in a DaCS Waa software controlled workbook 2YSTEM AUDIT Result 1 the Result worksheet s of the workbook Your results appear in User s Manual Refer to the DaCS User s manual which can New DaCs Workbook Ctrl N be accessed from the Support Tab in the a Resource Explorer for detailed information Load Template how this application interfaces with MS Open Workbook Ctrl O Excel Close Workbook Ctrl This additional DaCS drop down menu Save Workbook Ctrl 5 appears in MS Excel Some of the other Excel ne Save Workbook s Ctrl 4 drop down menus are also modified when Excel is opened in the MXP DaCS
212. rameter that lets you ignore events below the setting This lets you eliminate signals caused by debris DL filter A dichroic long pass optical filter that directs light in different spectral regions to different detectors Event A particle passing through the laser beam Export file XLS or TXT File containing selectable statistics and other sample information from each sample run FDA Abbreviation for fluorescein diacetate dye FITC Abbreviation for fluorescein isothiocyanate dye Flow cell A device through which particles pass in a stream of fluid one at a time through a laser beam GLOSSARY 2 PN 177579BD PN 177579BD GLOSSARY Flow cytometry A process for measuring the characteristics of cells or other biological particles as they pass through a measuring apparatus in a fluid stream Flow Cytometry Standard FCS The flow cytometry data file standard that provides the specifications needed to completely describe flow cytometry data sets within the confines of the file containing the data FlowPAGE A template for creating a report It can include data plots statistics and user attached text and or graphics Fluorescent light The emission of electromagnetic radiation that occurs when the emitting body absorbs radiation from some other source For example when a fluorescent dye is excited absorbs radiation it emits fluorescent light at a wavelength that is different from the wavelength of the
213. re than 5 from the mean integral signal channel number obtained within a period of 24 hours when the temperature does not vary more than 5 F from temperature at alignment Refer to the Flow Check fluorospheres package insert for details Stability for FL5 PC7 mean channel value of the integral signal intensity from the 488 nm laser using Flow Check 770 beads does not vary more than 5 from the average of the integral signal intensity mean channel values obtained within a period of 24 hours when room temperature does not vary more than 5 F from temperature at alignment Refer to the Flow Check 770 fluorospheres package insert for details Stability for FL4 APC mean channel value of the integral signal intensity from the red laser using Flow Check 675 beads does not vary more than 5 from the average of the integral signal intensity mean channel values obtained within a period of 24 hours when room temperature does not vary more than 5 F from temperature at alignment Refer to the Flow Check 675 fluorospheres package insert for details PN 177579BD LOG SHEETS WAN This appendix contains the following log sheets e Reagent Log e Action Log e Flow Set Fluorospheres Charts gt Daily Log for Instrument Standardization gt Establishing HV Total Gain Ranges e Flow Check Fluorospheres Charts gt Daily Log for Instrument Verification of Alignment and Fluidics gt Establishing Peak Position and HPCV Target Ranges Make photo
214. red from the FlowPAGE PN 177579BD USING MXP SOFTWARE B FLOWPAGE MENU Editing FlowPAGEs See also Creating FlowPAGES in the System Overview chapter in the Instructions For Use manual Select a Single Item Within FlowPAGE 1 Mouse over the item until you see the gt cursor 2 D the item and handles appear around the item Move a Selected Item Within FlowPAGE 1 Mouse over the item until you see the cursor and O hold and drag the selected item to its new position Select a Group of Items Within FlowPAGE 1 Click and hold the mouse button selecting just outside the group of items required 2 Drag the cursor until the marquee dotted outline completely encompasses the items you want to select 3 Release the mouse button Each item has handles around it PN 177579BD B 155 USING MXP SOFTWARE FLOWPAGE MENU 4 Mouse over the selected items til you see the P cursor and W hold and drag the selected items to move them 5 Click the right mouse button on the selected items to edit them Deleting Objects in FlowPAGE 1 W the object that is to be deleted Handles appear around the object 2 Press Delete Resizing Objects in FlowPAGE 1 D the object that is to be resized Handles appear around the object 2 Draga handle to the size required and release the mouse button B 156 PN 177579BD Additional FlowPAGE Formatting Options USING MXP SOFTWARE FLOWPAGE MENU Cl
215. red laser B 168 discriminator definition GLOSSARY 2 specifications 3 4 dissipation heat special requirements 2 2 dividers PRISM plot B 113 deleting B 115 dual parameter B 113 B 143 B 144 editing B 114 editing single parameter B 146 removing B 115 single parameter B 113 B 145 DL filters definition GLOSSARY 2 documentation for your instrument about your reference manual xxi conventions used xxii introduction to your manuals xxi using the manuals xxi dot plot coloring mode B 55 copy plot data B 96 copy plot image B 95 creating B 95 data source B 96 events B 97 apply to all plots B 98 no of events to show B 98 font B 99 labeling B 98 show axis labels B 99 show axis tick marks B 99 show plot title on output B 99 x and y axis labels B 99 drainage requirements 2 2 Duplicate Plot B 94 INDEX 8 edit copy B 59 cut B 59 delete B 61 FCS header attributes B 41 FlowPAGE align bottom B 162 align left B 161 align right B 161 align top B 162 change plot B 160 copy B 157 cut B 157 grow to largest B 160 paste B 157 reorder B 157 resize objects B 156 save as B 163 shrink to smallest B 161 space evenly across B 162 space evenly down B 163 text B 159 FlowPAGE fill B 157 FlowPAGE font B 157 FlowPAGE line B 157 font B 62 font FlowPAGE B 63 menu B 59 paste B 60 paste special B 61 PRISM plot dual parameter PRISM dividers B 144 single parameter PRISM divider B
216. rism Prism Scaling Control Format Plot The plot type with the current Gate is displayed in the plot window Ungated is the default B 110 PN 177579BD USING MXP SOFTWARE PLOTS MENU Data Source Prism Choose the File and Gate to be used for the Prism plot File Browse Button Gate Selection Apply Gate to All Plots Prism Scaling Control Use this option to set the Prism plot scaling Clip Channels Prism Plot Properties Automatic Maximum Value Set Maximum PN 177579BD B 111 B USING MXP SOFTWARE PLOTS MENU What is Prism Prism is a method of summarizing SOGOU oI x multi parameter surface marker data as a one dimensional Prism plot representing all possible subpopulations or phenotypes The Prism plot is a simple bar graph The height of each bar represents the count of each phenotype The count as a percentage j of all events or gated events is written adjacent or above each bar The prism plot J can be gated in which case the values 616 represent the percentage of gated prism events and the gate is indicated in the title bar How is Prism Created In single parameter analysis of surface marker data one or two regions are normally set to measure both Negative and Positive cells We can then look at a second parameter and calculate the 2 phenotypes from parameter 1 positive populations and the two phenotypes from parameter 1 negative populations four phenotypes in tot
217. rithmic and uncalibrated the values vary between 0 1024 1024 and span four decades For most other manufacturers the data varies between 1 10 000 spanning four decades B 23 21 CFR 11 MENU The 21 CFR 11 drop down menu only appears if you have the 21 CFR Part 11 option installed on your system The functions on this menu help you to be in compliance with the 21 CFR Part 11 Regulation Lock Software Use D Lock Software to manually lock the MXP software until you log back into the system This grays out all functions in the software except Log In Use File gt gt Log In to re enter the MXP software Only the current user or the System Administrator can log into the MXP software when it has be manually locked You need to lock the software whenever the instrument will be out of your control and you do not want to log out of the software These automated activities continue without interruption even when the software is locked e Acquisition e Listmode playback e Batch AutoMATOR e AutoSetup Change Password Use the Change Password feature to change the Password for your User ID Note After you change your password the System Administrator cannot view it The change password feature is not available during any of these automated activities Acquisition Listmode playback Batch AutoMATOR and AutoSetup PN 177579BD B 175 USING MXP SOFTWARE 21 CFR 11 MENU B 176 Procedure To change your password 1 21 CFR 11
218. ro Well Type Sample ld Dilution Replicates 6789 101112 1011112 DO as 5c 1 verify pro fm a T a e as 5c 2 settings pro as Sc 2l flow set pro 5 Se 2I neg contol pro Pae Pamen aaaea as 5c 2I verily pro Pine Tipe Volume pL Sample Requited pL efil compro s6wetNoma 5 z Cal Factor Mix Iterations Well priority Row O Column None 2 Z Ok Cancel Cleat Plate Help When you program a plate in the Plate Wizard and save the plate definition file tdf in the Trays directory a Worklist and a Plate Display are created and displayed Plate Display TSA S ot or Sy Oi 12 Ci M Me 3 eC LL VL GP Ya P Worklist 2C Cell Activation PRO FS Lin SS Lin FL1 Log FL2Log FL3Log FL4 Log A4 Cel Activation NEWtar heros Calibrator RA p s2 04 _00000003 004 LMD See also Acquisition Manager B 14 PN 177579BD USING MXP SOFTWARE B FILE MENU Sample Plate Analysis with MPL_DaCSAnalysis Add In or MPLAnalysis Add In Overview There are two Add Ins for use in Microsoft Excel supplied with MXP software e MPL_DaCSAnalysis Add In e MPLAnalysis Add In See the System Administrators Guide for instructions to install the MPL_DaCSAnalysis Add In and the MPLAnalysis Add In The Add In you use depends upon whether or not your system has the 21 CFR Part 11 option installed on it e Use the MPL_DaCSAnalysis Add In if you have the 21 CFR Part
219. rocedure Standard MXP Software Installed Note MXP software only saves one Histogram Plot in a single FCS histogram file Save Histogram File As Procedure With the 21 CFR Part 11 Option Installed 1 Highlight the Histogram Plot Current window 2 D File Save Histogram File As 3 Specify the name of the file Save HST File Save in HST hd i L 5c 11 as verify 45 56 19 3 8 Lymph FL1 Log v FL2 Log HST la 5c 11 as verify 45 56 19 3 8 Lymph 1 1 1 L 5c 11 as verify 45 56 19 3 8 Lymph FL1 Log v FL3 Log HST La 5c 11 as verify 45 56 19 3 8 Lymph a the desired directory m the L 5c 11 as verify 45 56 19 3 8 Lymph FL1 Log v FL5 Log HST La 5c 11 as verify 45 56 19 3 8 Ungate Save in list bo x L 5c 11 as verify 45 56 19 3 8 Lymph FL2 Log v FL4 Log HST L 5c 11 as verify 45 56 19 3 8 Lymph FL2 Log v FL5 Log HST g Filename 5c 11 as verify 45 56 19 3 8 Lymph FL2 Log v FL4 Log HST Save b the type of file in the Save as lt O ce type list box Saving As Version 2 Default Comments ERTE CHET E EE SEES c Inthe File name text box type the ERRET IN Eea desired File Name adjusted added regions gates 4 You must enter a comment before you can save a new version e Select an appropriate comment from the list of Default Comments OR e Type in a comment in the Comments for this version field Note Once a comment is entered the version number automatically appears in the Saving As Version field an
220. rosoft Windows documentation Font Eont farial E Arial Black T Arial Narrow T Book Antiqua T Bookman Old Style T Bookshelf Symbol 1 E Bookshelf Symbol 2 Effects _ Sample T Strikeout T Underline Ayy Color L Black X Script Westem jE This is a TrueType font This same font will be used on both your printer and your screen For True Type fonts you can generate both horizontal and vertical fonts for the Plots For non True Type fonts that do not have point sizes for both horizontal and vertical portrait and landscape Windows attempts to use a horizontal font for the vertical direction Under these circumstances choose another font or reload the font of your choice making sure the font sizes required are available in both portrait and landscape types Please see the Microsoft Windows documentation on loading new fonts See also Edit Font FlowPAGE Font PN 177579BD USING MXP SOFTWARE B EDIT MENU Edit Font FlowPAGE To change the fonts in FlowPAGE 1 Double click on a plot or text box to display the grap handles 2 Click the right mouse button on the plot or text box to display the properties dialog box 3 the Font tab 4 C the desired font font style size and color Underlined or strikethrough text can be shown C to set your new values 5 J once outside the plot or text box The cursor reverts back to the Chooser cursor ls and the handles
221. rs are photo multiplier tubes PMTs that have a 185 nm to 900 nm spectral range Signal Processing High voltage amplification minimum 250 up to 1 100 in increments of 1 for FLI FL2 FL3 FL4 FL5 Vernier gain fine amplification up to 1 000 labeled volts in increments of 1 for the following A change of 1 to 750 represents a 1 to 4 change in gain FS SS AUX Linear amplification gain by 1 0 2 0 5 0 7 5 10 20 50 75 100 200 500 or 750 for FS SS Linear amplification gain by 1 0 2 0 5 0 7 5 10 20 50 or 75 for FL1 FL2 FL3 FL4 FL5 Linear amplification gain by 1 0 2 0 5 0 7 5 10 20 50 or 75 for AUX Four decade digital logarithmic transformation of FS SS FL1 FL2 FL3 FL4 FL5 Note A scale of 0 1 to 1 000 is displayed on the plot axis for logarithmic parameters but the default statistics are based on an actual scale of 0 1024 to 1024 The displayed scale can be changed to 1 to 10 000 3 3 3 SPECIFICATIONS INSTRUMENT SPECIFICATIONS 3 4 e Fluorescence color compensation is available in 0 1 increments from 0 to 100 for FL1 FL2 FL3 FL4 FL5 e A discriminator maximum value of 1 023 is available for one of the following signals Only one discriminator can be specified for any one sample acquisition FS SS FL1 FL2 FL3 FL4 FL5 AUX Workstation The descriptions below are approved configurations Contact your Beckman Coulter Representative on current
222. rt 11 option installed on your system see the heading Resource Explorer for instructions how to check in these files for your use Panel Name Protocols Cleanse PNL Cleanse Bleach pro Cleanse Water pro Protocols 2c 1L pro 4c 2L pro QC 1L Flow Check pro 3c 1L pro 5c 1L pro QC 2L Flow Check pro 4c 1L pro 5c 2L pro HeNe Alignment pro 3 8 PN 177579BD SPECIFICATIONS SOFTWARE SPECIFICATIONS FCS Header Keyword Reference The Following FCS keyword information is included in MXP listmode and histogram file header section Those prefixed by a are FCS Standard defined keywords those prefixed by an or nothing are MXP defined keywords FCS Keyword Key value DATATYPE Single character defining base data type of binary data l 16 bit integer F IEEE 32 bit floating point D double precision 64 bit floating point A ASCII PAR Number of parameters MODE Single character defining mode of data L listmode C dual parameter correlated U single parameter uncorrelated PnB Number of bits allocated to store data for parameter n 8 means 256 channel data 16 means up to 64 k channel data PnR Channel resolution of parameter n for example 256 1024 and so on BYTEORD INTEL platform 1 2 Motorola platform 2 1 SNEXTDATA Byte offset into binary data for next data item FCS files can be saved as a pseudo linked lis
223. s or using quadrant markers four rectangular Regions set with only one point To speed up the analysis of data when a Single Parameter Histogram Region or a Rectangular or Polygonal Region is created it is also assigned to a Gate This assignment can be accepted or edited later The automatic assignment of Regions to Gate can be switched on or off if desired using Gates gt Automatic Gate Creation PN 177579BD B 131 USING MXP SOFTWARE ANALYSIS MENU B 132 Region Properties C on a region to make it active and then click the right mouse button on it to display the Region Properties Region Properties dialog box Re Calculate Autogate The Re Calculate Autogate option allows a return to AutoGating after manual manipulation of an Auto region See also Linear Region Linked Marker Create Multiple Linear Regions New Non Linked Marker Polygonal Region Quadrant Region Rectangular Region Create AutoGate Linear Region Create Multiple Linear Regions Linked Marker New Non Linked Marker Polygonal Region The Polygonal Region allows you to draw a free form Region with up to 100 points and can include horizontal acute angles on a dual parameter plot To draw a Polygonal Region in a dual parameter display see Create Polygonal Regions in the Instructions For Use manual Polygonal Region Properties Region Name Prime Automatic Region Name Positioning None Elliptical Contour Sensitivity Travel Intera
224. s connected to a 20 L waste container Dispose of the waste in accordance with your local environmental regulations and acceptable laboratory procedures WARNING Risk of biohazardous contamination if you have skin contact with the waste container its contents and its associated tubing The waste container and its associated tubing might contain residual biological material and must be handled with care Clean up spills immediately Dispose of the contents of the waste container in accordance with your local regulations and acceptable laboratory procedures The waste line supplied with the instrument can be connected to an open drain If you use an open drain mechanically secure the waste tube into the drain so the tube cannot accidentally come out of the drain This prevents spillage PN 177579BD INSTALLATION 2 SYSTEM CONNECTIONS 2 3 SYSTEM CONNECTIONS Power and Signal Cables Figure 2 1 or Figure 2 2 shows the interunit connections of the power and signal cables depending upon your Power Supply configuration Computer configuration may vary based on current configuration availability Note If the Cytometer is not connected to the Workstation when the Acquisition software is started the Help About screen displays an incorrect version number for the Cytometer Firmware Figure 2 1 Power and Signal Cable Connections with Voltage Specific Power Supply Cytometer penon FIBER OPTIC REC TO le XMIT
225. s the listmode file name extension Next File Name Displays the next listmode file name Once assigned in the Plate Wizard the well number of the sample is added to the front of this file name PN 177579BD B 47 USING MXP SOFTWARE FILE MENU Workspace Preferences Acquisition Options Sets the default Acquisition Options for MXP software during Acquisition allowing automatic Saving Printing and Exporting of Data B 48 Output Options Acquisition Manager Plot Display Gating Publish Default Log Scaling LMD File Name Acquisition Options User Info Cytometer Control Translucency Output Options VV Save LMD IT Save Histograms VV Save Protocol gt Publish J Print Plots T Print Statistics T Print FlowPAGES I Print FlowPAGES To PDF Print Plots To PDF Acquisition Manager TT Use Spaces Between Panels IT Continue AutoSetup on FlowSet HV Setup Failure Default Log Scaling Display as 1 10 000 Display as 0 1024 1 024 Cytometer Control Translucency Cancel Undo Apply Help See also Workspace Preferences Introduction Workspace Preferences LMD File Name Workspace Preferences User Info Workspace Preferences Plot Display Workspace Preferences Gating Workspace Preferences Publish PN 177579BD PN 177579BD USING MXP SOFTWARE FILE MENU Output Options The Output actions occur automatically at the completion of the acquisition of each sample
226. s you through creating or editing a panel PC7 Abbreviation for phycoerythrin cyanine 7 tandem dye pg picogram A unit of mass equal to 10 12 gram Photo multiplier tube PMT A light sensitive sensor that converts light energy into electrical current and generates a voltage pulse signal Pickup lens spatial filter assembly Collects side scatter and fluorescent light from only the sensing area of the flow cell and collimates it Plate Definition File A tdf file is created when you define and save sample plate information in the Plate Wizard The file contains the plate type and all the well assignments Plate Wizard A software screen that guides you through creating or editing a tdf file for sample plate assignments and worklist creation Pop up window A rectangular area that appears on top of the current screen displayed on the Workstation You must close the window before you can use the current screen again Positives analysis Analysis performed on the negative control to set regions automatically to exclude the negative population from the positives statistics Power Supply The system component that provides direct current power pressure and vacuum to the Cytometer Precision Ability of an instrument to reproduce similar results when a sample is run repeatedly Precision shows the closeness of test results when repeated analyses of the same material are performed Also known as reproducibility Contrast with
227. se manual Dual Laser Filter Block An assembled filter block configured as shown in Figure 5 6 in the Instructions For Use manual Single Laser Filter Kit The single laser filter kit includes two filters a 620 BP filter and a 645 DSP filter Dual Laser Filter Kit The dual laser filter kit includes two filters a 620 SP filter and a 615 DSP filter Software Options PN 177579BD 21 CFR Part 11 Option By providing audit trails and electronic signatures for files this option provides the tools for your data files produced by the MXP software to be in compliance with the requirements of the FDAs Electronic Records and Electronic Signatures Rule 21 CFR Part 11 DaCS software is supplied with the 21 CFR Part 11 option DaCS software provides the controls to help you bring your laboratory into 21 CFR Part 11 compliance for data files exported to Microsoft Excel from the MXP software 1 3 i USE AND FUNCTION REAGENTS AND QUALITY CONTROL MATERIALS 1 3 1 4 REAGENTS AND QUALITY CONTROL MATERIALS Beckman Coulter recommends these reagents or their equivalents All stated performance characteristics in this manual are based on the use of the Cytomics FC 500 MPL system with the following reagents Sheath Fluid In the Cytometer the sample is guided into a stream of sheath fluid to make the sample cells flow single file through the laser beam IsoFlow sheath fluid a nonfluorescent balanced electrolyte solution is made
228. setup If the sample is from a drug treatment protocol this could be a problem A 5 000 cell type A count may not be achieved before the sample well or tube is empty or RAM memory is full It is normal under these circumstances to set two counters one for the stop count and one for the maximum permissible count that is the maximum total event count Ifa stop count is used it is recommended that the stop count be set to a large number for example 100 000 events Thus with a drug treatment protocol if cell type A is less than 5 of the white cell count an acquisition would terminate on the total event count of 100 000 before a 5 000 cell cell type A count could be achieved PN 177579BD B 119 USING MXP SOFTWARE PLOTS MENU B 120 Setting a Stop Counter Note If a stop count is used on a plot that contains an AutoGate region the stop count is not exact 1 Highlight the plot on which you wish to set a stop counter 2 Plots Acquisition Save and Stops Data Source Histogram Scaling Stop and Save Font command to view the Acquisition Plot Acquisition Stop Condition Set up dialog box V Use Stop Condition Use Stop Condition Maximum Events 3000 Maximum Events Save Histogram Data in FCS Format I Save Histogram Data in FCS Format Cancel Help 3 J V Use Stop Condition and enter Maximum Events 4 To enable an FCS Histogram to be saved from this plot check the Save Histogram Data in F
229. sition Options User Info Opaque Region Labels Plot Display Gating Publish Print Plot Frames m General View Options m Baseline Offset Print Dots Black J Show in Region on New Plots Auto Show Overlay Legend Pane Tl Black Plot Backgrounds C On Show Overlay Statistics Pane F onsa Regon leidt eo Log Scale Density Plots Smooth Histograms Default Overlay Plot Background Color Default of Events on Dot Plot Baseline Offset Default Overlay Mode Default 3D Plot Mode Default Plot Size J Print Plot Frames J Print Dots Black Plot Creation Options T Show Overlay Legend Pane I Show Overlay Statistics Pane IV Log Scale Density Plots IV Smooth Histograms Default Overlay Plot Background Color H r Default Overlay h Mode Default of Events on Dot Plot Default Plot Sizee 0 50 100 C Small 180x180 E Or OscaLe Medium 225x225 4 C Large 300x300 Cancel Undo Apply Help See also Workspace Preferences Introduction Workspace Preferences LMD File Name Workspace Preferences Acquisition Options Workspace Preferences User Info Workspace Preferences Gating Workspace Preferences Publish B 51 B USING MXP SOFTWARE FILE MENU B 52 Show in Region When selected this automatically displays the percentage of events in a particular region All regions that have been created display this percentage Black
230. so Workspace Preferences Introduction Workspace Preferences LMD File Name Workspace Preferences Acquisition Options Workspace Preferences Plot Display Workspace Preferences Gating Workspace Preferences Publish Institute Enter the Institute name with a minumum of 3 characters It can be printed out as part of the statistics output Institute Street Enter the Institute street address It can be printed out as part of the statistics output Institute Town Enter the Institute town It can be printed out as part of the statistics output Sample Source This option allows you to enter information regarding where the sample came from B 50 PN 177579BD USING MXP SOFTWARE FILE MENU Project The name for the project is entered here Cell Type This labels the sample with the specific cell type being analyzed in that sample Experiment The name of the experiment being conducted is entered into this section Cytometer Serial Number The serial number from the specific Cytometer being used is displayed here and recorded in the listmode file as more than one Cytometer may be utilized in the same experiment You must be logged on as Administrator to enter the serial number Currently Logged On User The User ID of the currently logged on user is displayed here and recorded in the listmode file Workspace Preferences Plot Display PN 177579BD aes A Show in Region z Black Plot Backgrounds LMD File Name Acqui
231. software Sign Worksheets environment System Administration For use of the DaCS features such as Change Password electronic signatures separately set up a Change Passphrase User in the DaCS software for each MXP User ID About Dacs Disable Dacs Log Off Quit Dacs Audit Trails DaCS Software PN 177579BD When working in MS Excel files with DaCS software an Audit Trail is provided as a separate worksheet _AUDIT within the workbook See the DaCS User s manual for details of what information is audited These MXP software related activities are also audited The cell based auditing functions are disabled while these activities are in process e Workbook creation e Workbook naming e Statistics data insertion e Workbook saving e Workbook closing C 5 C 21 CFR PART 11 OPTION COMPLIANCE INFORMATION DaCS SOFTWARE Electronic Signatures DaCS Software See the DaCS Users manual for details of the electronic signature feature for workbooks Setting Up User IDs MXP Software User IDs e The MXP software System Administrator sets up the User IDs and passwords for MXP software e When you are logged into the MXP software and data is exported to MS Excel you are automatically logged into the DaCS software e Your MXP User ID appears at the top of the Excel screen Ed Microsoft Excel DaCs File Fi_20040115_180055 xls File Edit view Insert Format Tools Data Window DaCS Help OG E SR SBBS O o Qs Mom gt
232. stem agreement or to the computer program or system documentation for the terms and conditions of that license PN 177579BD xxiii INTRODUCTION COMPUTER PROGRAM STATEMENT xxiv PN 177579BD USE AND FUNCTION fl 1 1 SYSTEM COMPONENTS The system components are shown in Figure 1 1 Figure 1 1 Cytomics FC 500 MPL Flow Cytometry System 7579118B 1 MPL Mouse 2 Cytometer 6J USB box Keyboard 7 Computer 4 Monitor Power Supply PN 177579BD 1 1 USE AND FUNCTION SYSTEM COMPONENTS Cytometer This unit analyzes the sample It contains the internal sheath fluid and cleaning agent containers Power Supply This unit provides DC power pressure and vacuum to the Cytometer A separate 20 L waste container is also provided Workstation The Workstation runs the software that enables you to control the instrument It displays sample results and other information It consists of e A monitor e A computer with data storage devices e A USB box e A keyboard and a mouse Multi Plate Loader MPL The MPL is an automated sample loader for the instrument It accepts these sample plate types e 96 well plate normal or deep round or square wells e 24 well plate or e 40 tube holder supplied by Beckman Coulter This holds 12 x 75 mm plastic test tubes MXP Cytometer Software This contains the MXP Acquisition and Analysis software and is used when running the Cytometer This appli
233. t Passwords PN 177579BD All user accounts are password protected MEP Cytometer Startup Wizard Ej Follow your laboratory procedures for DD A R boni IE Administrator No creating and maintaining passwords m ii F Diane No 3E Theresa No QUO SR MXP Software MXP Software is supplied for General Use with FC 500 MPL Cytometers 1993 2004 Applied Cytometry Systems This program is protected by U S and International copyright laws as described in the About Screen and or product manual 2004 Beckman Coulter Inc dn Cancel Est Help Be sure to include the following criteria in your laboratory procedures concerning passwords and User IDs C 1 21 CFR PART 11 OPTION COMPLIANCE INFORMATION INTEGRITY OF ELECTRONIC DATA C 2 Administrator password It is vital that the Administrator Password is changed from the default password as soon as possible See the System Administrator Guide for additional details Note Be sure to maintain the Administrator password in a secure location If you lose or forget your Administrator password call your Beckman Coulter Representative Unique User ID password combination The System Administrator sets up each User ID and assigns an initial password The MXP software makes the User create a new password before logging in whenever the System Administrator assigns a password Once the User creates a new Password it cannot be viewed by the System Administrator
234. t of events on dot plot B 53 default 3d plot mode B 53 default overlay mode B 53 default overlay plot background color B 53 default plot size B 53 opaque region labels B 52 print dots black B 52 print plot frames B 52 show in region B 52 show overlay legend pane B 52 show overlay statistics pane B 52 smooth histograms B 52 workspace preferences B 51 plot options toolbar B 5 plots contour plot configuration B 101 creating contour plots B 106 density plots B 106 dot plots B 95 dual parameter PRISM dividers B 113 B 143 histogram plots B 100 legend plots B 116 PRISM plot dividers B 113 PRISM plot phenotypes B 112 PN 177579BD PRISM plots B 110 single parameter PRISM dividers B 113 B 145 data source color dot plot B 96 density plot B 107 histogram plot B 101 legend plot B 117 PRISM plot B 111 deleting PRISM plot dividers B 115 density plot configuration B 107 editing dual parameter PRISM dividers B 144 PRISM plot dividers B 114 single parameter PRISM dividers B 146 format options B 121 labeling color dot plot B 98 density plot B 109 histogram plot B 105 legend plot FCS information B 118 legend plot options B 117 gated B 118 total B 118 number B 118 show phenotype logic B 118 menu B 94 overlay plot linked marker B 137 non linked marker B 137 print B 43 PRISM view modify B 147 PRISM plot compatibility B 115 explained B 112 properties B 121 resolution co
235. t most often 0 DATE DD MM YY EXP Experiment name from Workspace Preferences PROJ Project name from Workspace Preferences OP Operator ID that is user name INST Institute name from Workspace Preferences FIL File name excluding path CYT Type of flow cytometer for example FC 500 MPL SMNO System defined run number that is a number which is incremented each time the flow cytometer is used to acquire data SRC SAMPLEID1 SYS MXP Intel Windows CELLS Cells name from Workspace Preferences BTIM Start time HH MM SS 24 hour clock delimiters ETIM End time HH MM SS 24 hour clock delimiters TOT Total number of events acquired PnV Voltage of sensor n PnGAIN Amplifier Gain setting of a sensor PnN Name of parameter n for example Manufacturer assigned name that is FS or FL1 LOG PnS User assigned name of parameter PnE Lin Log parameter calibration 0 0 means linear decades offset for example 4 0 1 means log PN 177579BD 3 9 3 SPECIFICATIONS SOFTWARE SPECIFICATIONS FCS Keyword Key value DFCiToj of FLi to subtract from FLj Y2KDATE Date in format YYYYMMDD SAMPLEID1 User defined string SAMPLEID2 User defined string SAMPLEID3 User defined string SAMPLEID4 User defined string PnZ Calibration for parameter n is active when this key value is ON PnX Linear calibration values in xx yy order for example xx ch
236. t to be displayed on the following row If an existing d is clicked the icon marker is removed and New Row is not forced In addition if Ctl is depressed while the File Gate or Parameters selection is made plots can be sorted by Gates within File and Parameters within Gates Files When is selected in the example Tile Special screen shown above the resulting plot display is shown to the right Arrange Icons This option is used to arrange minimized plot windows Plot windows can be minimized and placed anywhere on screen Window Arrange Icons to place all icons along the bottom of the screen Note This option is very useful when minimized plots have been lost on the desktop Choosing Window gt Arrange Icons places all the icons along the bottom of the MXP software Window Force Square Each window is squared so that horizontal and vertical sides are the same length That is aspect ration is forced to 1 1 This is used mainly for dual parameter plots Close All Choosing this option closes all MXP software windows PN 177579BD B 181 USING MXP SOFTWARE WINDOW MENU B 182 PN 177579BD 21 CFR PART 11 OPTION COMPLIANCE INFORMATION C 1 OVERVIEW The FDAs Electronic Records and Electronic Signatures Rule 21 CFR Part 11 outlines the requirements for submitting documentation in electronic form and the criteria for approved electronic signatures It defines the standards by which an organization can
237. t to save the protocols to your folder you must use the Save Protocol As Procedure With the 21 CFR Part 11 Option Installed feature and navigate to your folder and save the protocol B 37 B USING MXP SOFTWARE FILE MENU Save Protocol Procedure Standard MXP Software Installed Selecting this feature saves the Protocol in its current form Ctr S is the shortcut option as is the Save Protocol button Note If you open protocols from other Users folders and you want to save the protocols to your folder you must use the Save Protocol As Procedure Standard MXP Software Installed feature and navigate to your folder and save the protocol Save Protocol As Use this feature to save a protocol file under a different name or to a different folder Use the appropriate procedure depending upon whether you have the standard MXP software or also have the 21 CFR Part 11 option installed Select the appropriate link e Save Protocol As Procedure With the 21 CFR Part 11 Option Installed e Save Protocol As Procedure Standard MXP Software Installed Save Protocol As Procedure With the 21 CFR Part 11 Option Installed Use this procedure if you have the 21 CFR Part 11 option installed on your system 1 D File Save Protocol As 2 Specify the name of the file Save Protocol Save in AcquisitionProtocol Ad C Oa B AS 2C Settings PRO Boas de lly i i i Oe R AS 2C Flow SetPRO BP AS ACIL a the desired directory in the H
238. te count J Cells pL from the Analysis Select Results menu option Setting a CAL Factor for Absolute Counts The Absolute Count dialog box allows you to store the absolute count calibration particles Lot Number and Expiration Date information for reference use only Enter the CAL Factor to be used in the Plate Wizard when setting up the Worklist Note The number of events in the CAL region must be greater than 1000 otherwise absolute counts are not determined Absolute Count Calibration a xi Available Absolute Count Calibrators Lot Number ID pi 757 802 1027 20 Feb 2003 1000 21 Mar 2004 r Absolute Count Calibration Lot Number ID 756 802 Cal Factor 1000 Expiry Date far March z 2004 Day Month Year Add Repar Delete Manual Absolute Count Calibration Value 0 T Enable OK Cancel Help En PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Advanced Button If your System Administrator has assigned you access rights the Advanced button is selectable and you can enter and modify batch and count information Expiry Date Lot Number ID and CAL Factor can be entered here Add allows the addition of a new Lot Replace allows you to replace the selected information with updated values Delete removes the selected Lot from the list Setting a CAL Region Create a Region around the absolute count calibration particles the region MUST be named CAL Set the calibration
239. te worksheet containing the statistic information for each listmode file from the worklist e Experiment Table Format gt Creates one worksheet containing the statistic information of all of the listmode files from the worklist Export Bitmap Size Allows you to select the size of the Bitmap used in when exporting Log In This feature is only available if you have the 21 CFR Part 11 option installed on your system Use this feature to log into the MXP software if you are manually or automaticaly locked out of the system e See the heading Lock Software for details about manually locking the software e The system automatically locks you out of the software after a time interval of mouse or keyboard inactivity as defined by the System Administrator The System Administrator can also use the Log In feature to log into the MXP software when the system is manually or automatically locked However the User ID that was logged in before the lockout occurred is listed as the User ID for any activity the System Administrator performs while in the MXP software To ensure accurate Audit Trail information the System Administrator should log out as soon as possible after performing any needed actions in the MXP software Note If you reach the limit of password attempts allowed you are not able to log into the MXP software You must ask the System Administrator to unlock you and or reset your password before you can log into MXP software PN 1775
240. tegral B 137 x coordinate B 137 system status B 173 data rate B 174 elapsed time B 174 flow rate B 174 total events B 174 status page Cytometer settings B 171 stop acquisition B 119 counter maximum events B 121 minimum count B 136 save histogram data in FCS format B 121 use stop condition B 121 Cytometer B 172 surface markers precision 3 11 throughput 3 11 surface plot introduction B 110 System Excel worksheet C 5 system components l 1 connections 2 3 system components description Cytometer 1 2 MPL Multi Plate Loader 1 2 Power Supply 1 2 Workstation 1 2 system status status bar B 173 data rate B 174 elapsed time B 174 flow rate B 174 total events B 174 T tag number LMD file name B 47 TDF file extension 3 6 temperature ambient operating 2 2 test tube size 3 1 TESTFILE key value 3 10 INDEX 21 INDEX TESTNAME key value 3 10 threshold value displayed on Excel graphs B 15 throughput performance specifications 3 11 Tile Windows B 179 Tile Special Windows B 180 time LMD file name B 46 tomogram plot introduction B 110 toolbar acquisition manager B 7 AutoMATOR B 8 buttons B 4 customize command tab B 73 customize toolbars tab B 72 Cytometer B 9 definition GLOSSARY 6 file options B 5 FlowPAGE B 7 gate color options B 6 gate options B 6 help B 6 listmode playback B 10 plot options B 5 region options B 6 statistic options B 6 tools listmode
241. tems in the group grows to the size of the item which has the largest area B 160 PN 177579BD USING MXP SOFTWARE B FLOWPAGE MENU Shrink to Smallest 1 Select the items to be changed See Select a Group of Items Within FlowPAGE 2 E the button All items in the group shrink to the size of the item which has the smallest area Align Left 1 Select the items to be aligned See Select a Group of Items Within FlowPAGE 2 2S All items are now aligned to the item that was the furthest left in the objects selected Align Right 1 Select the items you wish to align See Select a Group of Items Within FlowPAGE 2 D Bal All items are now aligned to the item that was the furthest right of the objects selected PN 177579BD B 161 USING MXP SOFTWARE FLOWPAGE MENU Align Top 1 Select the items to be changed See Select a Group of Items Within FlowPAGE oo 2 J Edi All items are now aligned to the item that was the topmost in the section that was selected Align Bottom 1 Select the items to be changed See Select a Group of Items Within FlowPAGE 4 2 J E All items are now aligned to the item that was nearest the bottom of the objects that were selected Space Evenly Across 1 Select the items to be changed See Select a Group of Items Within FlowPAGE 2 iM All items are now horizontally spaced equally B 162 PN 177579BD Space Evenly Down USING MXP SOFTWARE FLOWPAGE
242. ter is deleted altogether from this plot and from any other plot that has this divider set Any associated Prism plot changes its format and has its statistics updated to reflect a smaller number of Prism divider s B 144 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Single Prism Divider The Analysis gt Create Prism Divider gt Single Prism Divider menu option allows a single Prism divider to be set on a single parameter plot Any displayed Prism plots is automatically updated with the current Prism statistics If the number of Prism dividers is changed during the divider creation the corresponding Prism plot also reflects this by changing its format according to the number of active Prism dividers Please refer to your Cytometer system manuals for further explanation on Prism parameter Note MXP Cytometer software only allows one Prism divider per parameter As a consequence modifying the Prism divider of any parameter on any plot automatically updates the divider of that parameter if it is displayed on any other plot This includes both single and single parameter plots See also Setting a New Single Prism Divider Editing a Single Prism Divider Setting a New Single Prism Divider 1 Highlight the required single parameter plot 2 W Analysis Create Prism Divider gt Single Prism Divider 3A Single Prism cursor is displayed in the current window 4 Position the cursor anywhere on the window click to sho
243. terest either for plot display or for sorting Gate Status Logic Editing 1 i Color Name Logic Expression 2 0 tne os oon Ungated events color AND oR XOR M Delet Gate and Logical Expression m De oe nor _ t _i ndo Gate Color Ca ea ee u New Delete Operands 3 New Gate 4 is highlighted so you can modify the name if desired 4 Create new logical expressions See also Gate Selection Gate Color Boolean Gating Gate and Logical Expression Shows the Gate name and the logical combination of Gates New Allows you to create a new Gate with a new combination of Regions Delete Delete erases any selected gates highlighted in the Create Modify Gates dialog box B 122 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Operands The Operands function allows you to insert a combination of regions into the gating equation using the Operands buttons Once a Region has been entered next to the new file name the Operands have to be utilized to enable a new combination of cells to be viewed The Operands work according to Boolean Gating Gate Selection The Gate option can be used once Gates have been created To apply a Gate to a plot 1 Click the right mouse button on the plot and O ES Format Plot 2 the Gate you wish to apply from the Gate drop down list box 3 J or press to accept the changes C or press to exit the dialog box without saving the changes P
244. ters The first ten characters of the protocol filename are used to generate the listmode filename for acquisition If the protocol name is longer then the name is truncated to 10 characters Sample ID 1 2 3 4 The Sample ID columns allow you to enter information about the particular sample being run When this box is checked in Workspace Preferences the information in any of these columns is included as part of the listmode file name Each Sample ID column corresponds to information entered in the Plate Wizard about the Sample Id Well Type Replicates and Dilution Note Be sure to verify manually entered sample IDs Well Loc This option is not available User ID When checked this includes the User ID as part of the listmode file name Run Number The system automatically generates an incrementing run number every time that a sample is run When this option is selected the run number is included as part of the listmode file name The easiest way to ensure unique file names are always generated If not selected the run number is still generated saved and can be accessed in the FCS Header section of the file Y2K Date When selected this option includes the date as part of the listmode file name The format used is the standard Year 2000 format YYYY MM DD showing when the sample was analyzed Time When this option is selected the time HH MM SS that the sample was analyzed is included as part of the listmode fil
245. the AutoGate algorithm fails during acquisition Pause on Error This option is not available in MXP software Abort Plate On Error The system automatically aborts the sample plate when an AutoGating error occurs Ignore Errors The system continues processing the Worklist when an AutoGating error occurs PN 177579BD B 55 USING MXP SOFTWARE FILE MENU B 56 Workspace Preferences Publish File References in Data Publish Data to Text File LMD FileName Acquisition Options Userinfo Publish Data to MS Excel men ea ia Switch to MS Excel on Publish File References in Data Export Bitmap Size Full Path amp Name Small 400x400 Publish to MS Excel Format great ener a Export Bitm iz c 800x800 port Bitmap Size I Publish Data to Text File Lae IV Publish Data to MS Excel IV Switch to MS Excel on publish Publish to MS Excel Format Cancel Undo Apply Help See also Workspace Preferences Introduction Workspace Preferences LMD File Name Workspace Preferences Acquisition Options Workspace Preferences User Info Workspace Preferences Plot Display Workspace Preferences Gating File References in Data Choose to reference the file name by Full Path and Name or by Name Only Publish Data to Text File Enable this checkbox to publish data to an ASCI text file with a file extension of TXT Publish Data to MS Excel This function enables you to publish data to a Microsoft Excel Wor
246. this command is active it informs you as to the length of time left in pause before the analysis resumes If this length of time is not required selecting the Run button again can restart the analysis e Continuous Pause This informs you that the processing is paused and can only continue if you restart the analysis manually by selecting the Run e button again Listmode Playback B 90 The Listmode Playback option allows for listmode playback with the runtime protocol panel or with a new protocol panel This option also allows listmode compensation of 20 bit data files in a fully automated manner replicating the procedure used during analysis Compensation settings may be verified and manually adjusted if required The Options tab allows setting of default options during listmode playback Note If you have the 21 CFR Part 11 option installed the Open Playback Panel Files dialog box accessed by contains the Show Archived checkbox and the Versions button The Save Playback Panel Files dialog box accessed by contains the Comments fields See also Open Protocol Procedure With the 21 CFR Part 11 Option Installed Save Protocol As Procedure With the 21 CFR Part 11 Option Installed PN 177579BD USING MXP SOFTWARE B TOOLS MENU Single File Mode DBW Ux n ti Seo ale Single File Mode List Comp Setup Options Panel Filename Compensation Filename ___Protocots Ron ____LmDFie Crp status Save This
247. to the MXP softwares BCAGP database A file can be checked into the BCAGP database by simply opening the file within the MXP software Once an uncontrolled file is checked in the file receives a version number and becomes a controlled file See Controlled Files in the Using MXP Software chapter for additional details The LMD file is an unchangeable record Each time you open an LMD file embedded with the runtime protocol for reanalysis it displays the original analysis regions and results as when collected along with the User ID and date and time stamp If you edit and display new results for an LMD file on the screen and then you save these changes an incremental version number of the file is saved with the changes made The LMD file that was initially acquired retains its own version number and is not changed Whenever a FlowPAGE or Workspace print is saved as a PDF file for a subsequent analysis the PDF file is saved with an incremental version number as well See also Save Listmode File As Procedure With 21 CFR Part 11 Option Installed Save Histogram File As Procedure With the 21 CFR Part 11 Option Installed Save Protocol Feature With the 21 CFR Part 11 Option Installed Save Protocol As Procedure With the 21 CFR Part 11 Option Installed 21 CFR Part 11 Option Uncontrolled and Controlled Files Windows Operating System The System Administrator should use the security and access features provided by Windows Operating System
248. tware MXP Software is supplied for General Use with FC 500 MPL Cytometers 1993 2004 Applied Cytometry Systems This program is protected by U S and International copyright laws as described in the About Screen and or product manual mou 2004 Beckman Coulter Inc Password 1 Admin Cancel Exit Help Note The System Administrator should refer to the System Administrator s Guide for detailed instructions about this screen and the System Administrator tools After the default Plate Display appears the Startup Wizard screen appears allowing Password entry and access to the MXP software On the initial Startup after installation only the Administrator User ID is available Some User IDs may appear on the initial Startup after installation if the System Administrator migrated these User IDs from a previously installed version of the MXP software All User IDs must first be set up by the System Administrator before they can log into the MXP software The Startup Wizard screen displays the User IDs of all those who have access rights to MXP software and whether they are logged in to the system or not If you have been previously set up on the system highlight your User ID and enter your Password The first time you log in after you were set up by the System Administrator the system prompts you to change your Password at the Password Change dialog box that appears The System Administrator cannot view any password you create
249. u will receive the most current information bulletins concerning the product If you purchased this product from a third party and would like further information concerning this topic call your Beckman Coulter Representative PN 177579BD REVISION STATUS n Issue A Initial Issue 8 03 MXP Software Version 1 0 Initial issue for customer distribution Issue B 11 04 MXP Software Version 2 0 Complete Revision Updated and new information was added for variable sample aspiration and mixing cycles in the Plate Wizard The Auto Startup function was deleted The optional 21 CFR Part 11 feature was added Issue BA 05 08 MXP Software Version 2 2 Updated or new information was added on pages 2 2 2 3 3 2 3 4 C 2 and C 3 Issue BB 03 09 MXP Software Version 2 2 Updates were made to pages 3 3 B 167 B 167 B 167 and C 2 Issue BC 05 10 Software Version 2 2 Updates were made to the company corporate address Issue BD 10 11 MXP Software Version 2 2 Analysis or Cytometer Additions changes or deletions were made at these locations xxi 3 3 B 1 B 37 B 53 B 150 B 152 B 166 C 3 and GLOSSARY 3 Note Changes that are part of the most recent revision are indicated in text by a bar in the margin of the amended page This document applies to the latest software listed and higher versions When a subsequent software version changes the information in this document a new issue will be released to the Beckman Coulter
250. uction B 154 menu B 154 move selected item B 155 paste objects B 157 print B 43 reorder objects B 157 resize objects B 156 select a group of items B 155 select single item B 155 toolbar B 7 PN 177579BD Flow Set 675 fluorospheres log sheets A 1 protocols 3 7 3 8 Flow Set 770 fluorospheres log sheets A 1 protocols 3 7 3 8 Flow Set fluorospheres description 1 5 log sheets A 1 protocols 3 7 3 8 See also controls fluidics systems check stability 1 5 fluorescence color compensation specifications 3 4 HPCV specification 3 11 resolution performance specifications 3 11 sensitivity performance specifications 3 12 standardize intensity 1 5 fluorescent light FL definition GLOSSARY 3 sensor definition GLOSSARY 3 fluorospheres protocols 3 7 3 8 font color dot plot B 99 edit B 62 edit FlowPAGE font B 63 force square Windows B 181 format FCS 3 5 overlay plots axis and tick mark labels B 110 plot options B 110 x axis labels B 110 PDE 3 5 plot B 121 forward scatter FS definition GLOSSARY 3 HPCY specification 3 11 resolution performance specifications 3 11 sensor definition GLOSSARY 3 freeze frame creating B 103 deleting overlay B 104 histogram plot configuration B 102 introduction B 102 FS See forward scatter FS PN 177579BD INDEX G gain definition GLOSSARY 3 settings Cytometer Control B 170 gate automatic creation B 54 color options toolbar B 6 create
251. um concentration 3 1 preparation 3 1 requirements 3 1 source user info B 50 Sample ID definition GLOSSARY 6 do not edit when using Add In B 20 edit in acquisition manager B 68 sample tube See test tube save all FCS format files B 31 histogram file as B 32 listmode file as B 28 protocol B 37 protocol as B 38 scaling histogram plot B 105 automatic maximum value B 105 clip channels B 105 set maximum B 105 overlay plots B 110 PRISM plot B 111 scatter sensitivity performance specifications 3 12 scroll bar definition GLOSSARY 6 SD standard deviation definition GLOSSARY 6 select definition GLOSSARY 6 select results B 148 advanced statistics configuration B 151 automatic statistic percentiles B 151 differences between log log and log lin statistical calculations B 151 log lin B 151 INDEX 20 log log B 151 current file name B 149 FCS information B 149 statistic type B 149 sensitivity 3 12 definition GLOSSARY 6 performance specifications 3 12 sensors fluorescent light definition GLOSSARY 3 forward scatter FS definition GLOSSARY 3 specifications 3 3 types 3 3 set maximum scaling histogram plot B 105 setting a Cal factor B 152 advanced button B 153 settings get cytosettings from LMD file B 171 get cytosettings from protocol B 171 import CMP file settings B 42 protocols 3 7 3 8 setup mode Cytometer Control B 166 show in region B 52 dot plot axis labels B 99 dot plot
252. umatic Tubing 2 5 INSTALLING MXP SOFTWARE 2 6 SPECIFICATIONS 3 1 3 1 Ad 3 3 SAMPLE REQUIREMENTS 3 1 INSTRUMENT SPECIFICATIONS 3 1 Dimensions 3 1 Installation Category 3 1 Noise 3 1 Cytometer 3 1 Flow Cell 3 1 Flow Rate 3 2 Lasers 3 2 Argon Laser Power 3 2 Beam Shaping Optics 3 2 Argon Laser Beam Spot Size 3 2 Solid State Laser Beam Spot Size 3 2 Optical Filters 3 2 Sensors 3 3 Signal Processing 3 3 Workstation 3 4 Computer 3 4 Data Storage 3 4 Interfaces 3 4 Input Devices 3 4 Monitor Options 3 4 SOFTWARE SPECIFICATIONS 3 5 Data Output and Compatibility 3 5 Setup Mode 3 5 Acquisition 3 5 Regions 3 5 Listmode Analysis 3 6 MXP Default File Extensions 3 6 Standard Default Files and File Extensions 3 6 Default AutoSetup Protocols and Panels Administrator 3 7 Default AutoSetup Protocols and Panels All Users 3 8 Default Generic Panels and Protocols All Users 3 8 FCS Header Keyword Reference 3 9 PN 177579BD 34 PERFORMANCE SPECIFICATIONS 3 11 Carryover 3 11 Data Acquisition Throughput 3 11 Precision for Surface Markers 3 11 Resolution 3 11 Forward Scatter 3 11 Fluorescence 3 11 Sensitivity 3 12 Scatter 3 12 Fluorescence 3 12 Stability 3 12 Day To Day 3 12 Within Day 3 12 A LOG SHEETS A 1 B USING MXP SOFTWARE B 1 B 1 B2 B 3 BA B 5 B 6 Ey B 8 PN 177579BD WELCOME TO MXP SOFTWARE B 1 CHECK FOR BCAG P DATABASE ICON B
253. utomatically assigned an alphabetical region name and a number for each quadrant See also Create Quadrant Regions and Repositioning a Quadrant Region in the Instructions For Use manual Quadrant Region Properties Region Name x X and X Coordinate PTE Positives Analysis X Coordinate 24 502 Y Coordinate jpa 59 Positives Analysis I Active Percent Positive E Cancel Help X and Y Coordinate Set the region to defined position by entering X and lower Y coordinates for the region Interactive Quadrant Region Editing When a Region s position is changed any Gates and statistics derived from the Region are automatically updated and each plot is redrawn to reflect the changes in the Gate When a Region is deleted any Gate or statistics that used the Region are also automatically deleted When a Gate has been deleted any plot which used the deleted Gate is reset to Ungated MXP software allows you to reposition an already created Region Elliptical AutoGate Create an Elliptical AutoGate around the selected population on a plot See also Create AutoGate in the Instructions For Use manual Contour AutoGate Create a Contour AutoGate around the selected population on a plot See also Create AutoGate in the Instructions For Use manual PN 177579BD B 135 USING MXP SOFTWARE ANALYSIS MENU B 136 Linear Region Insert a Linear Region See also Create Linear Regions in the Instructions For Use manual Linea
254. ve information B 137 Multi User Sign On B 2 MXP software defined keywords 3 9 file extensions 3 6 files written in ASCII 3 6 installation 2 6 negative control protocol to use in AutoSetup 3 7 network server option description 1 3 neutral density ND1 filter See filters new acquisition protocol B 12 panel B 13 plate B 14 protocol B 11 new panel B 13 noise specification 3 1 PN 177579BD INDEX non linked marker creating B 137 Not a Valid Worklist B 1 0 opaque region labels B 52 open drain 2 2 histogram B 30 listmode file B 23 protocol B 34 optical beam shaping 3 2 systems check stability 1 5 options dual laser filter block 1 3 dual laser filter kit 1 3 filter kit 1 3 hardware 1 3 legend plot B 117 gated B 118 total B 118 number B 118 show phenotype logic B 118 listmode playback B 93 Printer 1 3 second red laser 1 3 single laser filter block 1 3 single laser filter kit 1 3 software 1 3 Workstation network server 1 3 output options acquisition options B 49 AutoMATOR B 89 output txt file plate date display 3 6 overlay plot background color B 53 general axis and tick mark labels B 110 plot options B 110 x axis labels B 110 insert auto text B 83 insert histogram B 83 insert plot annotation B 83 linked marker B 137 non linked marker B 137 scaling B 110 INDEX 15 INDEX P panel wizard definition GLOSSARY 5 description B 13 panels
255. vious well or tube in a panel This option loads region positions from the protocol selected in the Use regions from this protocol field of the Panel Wizard E Region Filename Displays the name of the protocol selected in the Use regions from this protocol field of the Panel Wizard See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST B 66 PN 177579BD Acquisition Manager Parameter Names Parameter Names USING MXP SOFTWARE B VIEW MENU Column headings displaying the default parameter names from the protocol or panel See Edit FCS Header Attributes for a way of saving parameter names in a protocol prior to acquisition The Parameter Names can be customized and edited in the Panel Wizard In this example the columns are numbered P1 to P6 The actual number present is determined by the number of parameters selected in the protocol the maximum number you can have is 16 See also Acquisition Manager Worklist Columns Available ACQUISITION MANAGER TOOLBAR CREATING NEW PLATE WORKLIST Acquisition Manager Instrument Settings and Filename Instrument Settings CD10 FITC CD19 PE CD45 ECD Three state toggle displayed as icons or bitmaps These states being Loads the instrument cytosettings from the protocol 4 Carries the instrument cytosettings through from the previous acquisition Note When using the Instrument Settings column of
256. w B 122 Delete B 122 PN 177579BD xiii CONTENTS Operands B 123 Gate Selection B 123 Gate Color B 124 Boolean Gating B 125 Gate Logic B 125 Color Blend Mode Introduction B 127 Modify Color Blend B 128 Gate Names B 129 Enable B 129 Blend Color B 129 Name B 129 Color B 129 Logic B 129 Create Gate B 129 Modify Color Precedence B 130 Advanced Precedence B 131 Regions Introduction B 131 Region Properties B 132 Polygonal Region B 132 Polygonal Region Properties B 132 Region Name B 133 Prime B 133 Automatic Region Name Positioning B 133 AutoGating B 133 None B 133 Elliptical B 133 Contour B 133 Sensitivity B 133 Travel B 133 Minimum Count B 133 Polygonal Region Format Tab B 134 Interactive Polygonal Region Editing B 134 Rectangular Region B 134 Rectangular Region Properties B 134 Interactive Rectangular Region Editing B 135 Quadrant Region B 135 Quadrant Region Properties B 135 X and Y Coordinate B 135 Interactive Quadrant Region Editing B 135 Elliptical AutoGate B 135 Contour AutoGate B 135 Linear Region B 136 Linear Region Properties B 136 Lower Limit and Upper Limit B 136 Target Region for Automatic Setup B 136 Positives Analysis B 136 Interactive Linear Region Editing B 136 Status Bar Linear B 137 X Coordinate B 137 xiv PN 177579BD Integral B 137 Create Multiple Linear Regions B 137 Interactive Multi Linear Region Editing B 137 Linked Marker B 137 New N
257. w the prism divider lines If is pressed at this point the operation is abandoned 9 Move cursor to position crosshair then click to set position PN 177579BD B 145 USING MXP SOFTWARE ANALYSIS MENU Editing a Single Prism Divider Editing an existing Single Parameter plot with a set of single Prism dividers displayed essentially consists of creating a new single Prism dividers When the second single Prism dividers are defined the original is deleted 1 Highlight the chosen single parameter plot click on the title bar 2 Mouse over the Prism divider to get the tor cursor 3 hold to set the new divider The old divider is deleted and the display is once again updated to show only one single Prism divider Any associated Prism plot is updated with the new Prism statistics 4 To delete Prism divider on the divider to make it active and press Delete The Prism divider for the associated parameter is deleted altogether from this plot and from any other plot that has this divider set Any associated Prism plot changse its format and have its statistics updated to reflect a smaller number of Prism dividers B 146 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU View Modify Prism Under most circumstances Prism is modified graphically by using Dual or Single Prism Divider cursor operations Occasionally it is necessary to modify the Prism dividers numerically to reproduce exactly the setup of another e
258. website For labeling updates go to www beckmancoulter com and download the most recent manual or system help for your instrument REVISION STATUS iv PN 177579BD WARNINGS AND PRECAUTIONS REVISION STATUS iii INTRODUCTION xxi USING YOUR Cytomics FC 500 MPL MANUALS xxi ABOUT THIS MANUAL xxi CONVENTIONS xxii To Choose A Command With The Keyboard xxiii Dialog Box xxiii Description of Reporting Units xxiii GRAPHICS xxiii COMPUTER PROGRAM STATEMENT xxiii USE AND FUNCTION 1 1 1 1 SYSTEM COMPONENTS 1 1 Cytometer 1 2 Power Supply 1 2 Workstation 1 2 Multi Plate Loader MPL 1 2 MXP Cytometer Software 1 2 Microsoft Excel Software 1 2 1 2 OPTIONS 1 3 Hardware Options 1 3 Printer 1 3 Workstation Network Server 1 3 Second Laser 1 3 Filter Kit 1 3 Single Laser Filter Block 1 3 Dual Laser Filter Block 1 3 Single Laser Filter Kit 1 3 Dual Laser Filter Kit 1 3 Software Options 1 3 21 CFR Part 11 Option 1 3 1 3 REAGENTS AND QUALITY CONTROL MATERIALS 1 4 Sheath Fluid 1 4 Cleaning Agent 1 4 Quality Control Materials 1 5 1 4 MATERIAL SAFETY DATA SHEETS MSDS 1 5 INSTALLATION 2 1 2 1 DELIVERY INSPECTION 2 1 PN 177579BD CONTENTS vi pa 23 p SPECIAL REQUIREMENTS 2 1 Space and Accessibility 2 1 Electrical Input 2 2 Ambient Temperature and Humidity 2 2 Heat Dissipation 2 2 Drainage 2 2 SYSTEM CONNECTIONS 2 3 Power and Signal Cables 2 3 Waste and Pne
259. when two or more events are to be displayed at the same location 1 D Analysis Modify Color Precedence to view the Color Precedence Gates tab dialog box C Color Precedence 6 6 hCl i Gates A 2 the required gate so as to ates Advanced highlight the gate name ONLY Cancel Apply Help 3 Drag and Drop individual Gates into the required order of precedence Those with the highest precedence are at the top of the list See also Workspace Preferences Gating Advanced Precedence B 130 PN 177579BD USING MXP SOFTWARE B ANALYSIS MENU Advanced Precedence Advanced Precedence allows you to assign xi particular Gate colors to specific plots Gates Advanced Plots Precedence Gates This is of particular interest if different plots gt r AANA A LOG CD4 AD1 Include O C are gated on completely different criteria FL2LOGCD4ADI v SS 55 Excludes g and the color from one gate therefore has no relevance to another plot or file Regions Introduction A Region identifies a specific population of cells Up to 256 regions total can be set on any combination of Parameters 32 of these are usable as Gates Regions are drawn on a Single Parameter Histogram by setting the position of an upper and lower boundary Region markers Regions are drawn on a 2 Dimensional Dot Plot by enclosing a subpopulation of events with rectangles polygons nonrectangular Region
260. x allows the full file path for each file gt 01_Well 01 Specimen R_1 Undefined vandy 262 LMD 06 05 03 15 19 46 03_Well 03 Specimen R_1 Undefined vandy 263 LMD 06 05 03 15 21 14 05 Well 05 Specimen R_1 Undefined vandy 264 LMD 06 05 03 15 22 42 O7_Well 07 Specimen R_1 Undefined vandy 265 LMD 06 05 03 15 24 10 039_Well 09 Specimen R_1 Undefined vandy 266 LMD 06 05 03 15 25 36 11_Well 11 Specimen R_1 Undefined vandy 267 LMD 06 05 03 15 27 04 13_Well 13 Specimen R_1 Undefined vandy 268 LMD 06 05 03 15 28 32 15_Well 15 Specimen R_1 Undefined vandy 269 LMD 06 05 03 15 30 02 t 17_Well 17 Specimen R_1 Undefined vandy 270 LMD 06 05 03 15 31 28 19_Well 19 Specimen R_1 Undefined vandy 271 LMD 06 05 03 15 32 54 iJ 21_Well 21 Specimen R_1 Undefined vandy 272 LMD 06 05 03 15 34 20 23 Well 23 Specimen R_1 Undefined vandy 273 LMD 06 05 03 15 35 48 Cancel Help I Show file path AutoSetup Application Definition This option is not available with MXP software AutoSetup Scheduler This option is not available with MXP software B 19 PLOTS MENU Plots Introduction MXP software can display eight Plot types and the Legend for Color Gated Plots When a Plot is displayed click the right mouse button to display further menu options Color Dot Plot Histogram Plot Contour Plot not available Density Plot Overlay Histogram Plots not available Tomogram Plot not available Surface Plot not available Prism Plot Legend Duplicate P
261. x64 e Levels 2 3 and 4 0 11 0 33 and 1 1 at a resolution of 128x128 e Level 5 1 1 processed peak at a resolution of 256x256 Selecting 0 11 creates a larger region whereas selecting 1 1 creates a smaller radius region around the target population Travel Input a maximum value the contour AutoGate region travels from the mean The Minimum value is 25 Maximum is 250 channels Use this option to slightly adjust the AutoGate from sample to sample if slight changes in lysis cause the target population to move slightly This option is not available for elliptical AutoGates Minimum Count Check this option and enter a number in the Count field to enable the minimum count function This overrides an existing stop count to ensure a minimum number of events within a particular region are collected The actual stop count in this case is approximate but is always in excess of the minimum count value This is limited by the Duration and Total Events B 133 B USING MXP SOFTWARE ANALYSIS MENU B 134 Polygonal Region Format Tab Specify the Color and line Weight for the xj Polygonal Region boundary Region Properties Fomat Line Automatic Color ae Custom Weight Cancel Help Interactive Polygonal Region Editing When a Region s position is changed any Gates and statistics that use the Region are automatically updated and each plot is redrawn to reflect the changes in that Region When the
262. xperiment View Modify Prism allows you to do this 1 O Analysis View Modify Prism The Prism Setup dialog box is displayed C View Modify Prism xX 2 the parameter whose divider you List Mode Parameter wish to edit CD3ECD CD4 RD1 CDBFITC FS s Cancel Help 3 Place a check in the box next to the Listmode Parameter value you wish to enable Note If no check is in the checkbox the particular value is automatically set to OFE This switches off the prism divider and deletes it from any displays that were using it If a prism divider is switched off any active prism plots and plots with prism divider displays also update them to reflect the modified number of prism parameters in use 4 O the value that you wish to change and enter the required value within the range 0 to 1023 5 D to accept all changes to abandon all changes PN 177579BD B 147 USING MXP SOFTWARE ANALYSIS MENU Select Results B 148 MXP software gives the flexibility of generating many different statistics and these are produced using combinations of Regions and Gates A Gate is used to select events that may potentially be counted while Regions mark subpopulations and determine whether an event is actually counted All Regions and Gates can be named according to your preference allowing easy recognition of your results The statistics calculated and displayed depend upon those selected in the current protocol If
263. xt FlowPAGE enables custom formatting of multiple pages of flow cytometer information without the need to swap between other third party applications The automatic header information appears as XXXXs on the analysis FlowPAGE reports when viewed in the MXP software These automatic header fields display the Listmode file Keyword information only when the FlowPAGE reports are printed Once a FlowPAGE is created the contents of its plots and statistics windows are automatically updated to reflect new listmode data or configurations made in the MXP software analysis Listmode file names can become very long depending upon the naming options you have selected If these long Listmode file names result in the statistics boxes overlapping you may need to reformat the FlowPAGE to view all of the results All FlowPAGEs are saved with a Protocol this retains the link between the Plots you display and how you want to format the output to the Printer Note FlowPAGEs are not updated during Data Acquisition in live time FlowPAGEs are updated at the conclusion of acquisition or after reading a new data file from disk or editing Regions Gates or Statistics or resizing FlowPAGEs Note Occasionally when using a FlowPAGE one or more items cannot be deleted from a page If this occurs the items can be cleared from the FlowPAGE by selecting Files Save As to save the protocol with a new name If you reload this new protocol the items are now clea
264. y any provision of any purchase order receipt acceptance confirmation correspondence or otherwise unless Beckman Coulter specifically agrees to the provision in writing This agreement is governed by the laws of the State of California i B COULTER TRADEMARKS co The Beckman Coulter stylized logo and COULTER CLENZ are trademarks of Beckman Coulter Inc and are registered in the USPTO All other trademarks service marks products or services are trademarks or registered trademarks of their respective holders PN 177579BD Cytomics FC 500 MPL with MXP Software Documentation Reference PN 177579 Special Procedures PN 177580 Instructions For Use PN 624813 System Administrator s Guide PN 628899 Copyright Beckman Coulter Inc 2008 2011 All Rights Reserved Use and Function Installation Specifications Log Sheets Using MXP Software 21 CFR Part 11 Option Compliance Information e Glossary Index Cleaning Procedures Replace Adjust Procedures Index System Overview Daily Routine Quality Control e Running Samples Operation Principles Troubleshooting How To e Sample Plate Sizes Index Installation Administrator Tools With 21 CFR Part 11 Option e Administrator Tools Standard MXP Software Index Come visit us at www beckmancoulter com COULTER
265. ys Runtime Protocol if the current file is exactly as at the end of acquisition if any changes are made post acquisition and the LMD file is resaved the new file displays New Protocol RATIONUMERATORMUX Internal Mux address of the parameters used for the Ratio Parameter Numerator 3 10 PN 177579BD SPECIFICATIONS PERFORMANCE SPECIFICATIONS FCS Keyword Key value RATIODENOMINATORMUX Internal Mux address of the parameters used for the Ratio Parameter Denominator PANEL Name of the panel file used for acquisition of the sample ACQUISITIONPROTOCOLOFFSET Defines where in the LMD file the Runtime Protocol is stored the value displayed is number of bytes from start of file to the start of the protocol section ABSCALFACTOR Defines the calibration factor of the beads used with this sample if absolute calibration particles are used TRAYID Plate ID Name tdf file name TRAYROWCOL Well location designated by both a row letter and a column number Example A1 E40 Corresponds to the well location displayed on the Plate Wizard TRAYANALYSIS Sequential analysis code number assigned by the instrument to the plate 3 4 PERFORMANCE SPECIFICATIONS Carryover Scatter and fluorescence carryover is less than 1 from one specimen to another when the number of gated events is 10 000 or greater Data Acquisition Throughput Acquisition throughput is dependant on many variables i

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