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FluorChem E System User Guide

1. Select the Copy button from the dialog box When the copy is complete the following screen will display Copy to USB flash drive x Copy complete now safe to remove drive Sirs NOTE Do not remove the USB drive from the USB port until the copy process has completed or is cancelled The Copy command will automatically copy a folder to the USB drive The folder name will be the same as the fcz file and will contain the following files FluorChem E system fcz file This file can be used for image analysis in AlohaView software 16 bit png image file s These full bitmap files are ideal for quantitation and use in print publica tions A 16 bit ong file is provided for each channel 8 bit per channel png image file This smaller sized file is ideal for use in documentation and pre sentations as well as for e mailing This ong file is the overlaid multi channel image or single channel image based on the selection in the Exposures page The Title information entered by the user in the Image Info box will be used for both png file names Example png file names for an image with 2 channels would be New_Prep_Method_8bit ong New_Prep_Method_1_16bit ong and New_Prep_Method_2_16bit ong Channel numbers are appended to 16 bit ong file names FluorChem E System User Guide Copying Printing and Deleting Images page 69 NOTE If Title information is not entered the fcz file name will be used as the png file names
2. Tite 2011 01 19_17 03 35 User Project Tags Notes MAT je RR Tey pe pall fee PP Prot Se DA pe eS AY ee od eke shift Zal eel Get We B fey IM del 123 return The image title default is the date and time of acquisition and can be edited by the user Enter text in the title user project tags and notes fields as desired using the touchscreen keyboard FluorChem E System User Guide page 60 Chapter 6 Image Review When text entry is complete select Save Information entered will be saved with the final image and will also be searchable in the Browse page of the Images Screen Select Cancel to exit the Image Info page Channel Image Display and Overlay In the Review Screen image data for all channels acquired can be viewed individually or overlaid Channel overlay is ideal for visualization of colorimetric markers on chemiluminescent blots and chemiluminescent blots at various exposure times stacking Channel image data can be displayed or hidden by selecting the Image Show Hide button next to the Channel Settings box The button can be toggled between Show arrow only and Hide red line through arrow Image Show lt q Channel l Image Hide _ gt gf gt Channel2 NOTE When multiple channels are acquired channel image data will be displayed overlaid by default when the Review Screen is presented at the completion of acquisition Viewing Image Data for One Channel To v
3. page 130 Renaming Lanes Chapter 11 Analysis in Digital Darkroom Lanes can be renamed by selecting the lane number in the table and typing into the text box Lane Lane 1 73883 red Select Lane number Position Intensity 33 3 166738 30 0 62470 33 8 547217 37 4 366166 39 8 43586 48 9 32317 52 5 37722 29 7 39135 35 0 474025 39 3 19454 32 1 21663 8 2 83183 15 8 82285 25 4 75950 34 3 176731 52 8 236280 67 6 148718 89 4 191877 94 2 206699 Adding Molecular Weight Ladders To add a molecular weight ladder check the box next to Ladder Lane in the Analysis section Select the lane number your ladder is located in from the drop down menu FluorChem E System User Guide Total 100 0 5 7 Assigning Molecular Weight Ladders page 131 Analysis Define Lanes W Ladder Lane rs oO me w M Assigning Molecular Weight Ladders Once a ladder lane has been selected the molecular weight can be assigned by entering the molecular weight values directly into the table To do this click on the cell under MW that corresponds to the correct molecular weight and type After the values are entered the remainder of the table will automatically update FluorChem E System User Guide page 132 Chapter 11 Analysis in Digital Darkroom Lane MW Intensity Total 7 Lane 1 36 166738 100 0 Lane 2 39 73883 100 0 3 58 62470 5 7 33 547217 50 2 g 365166 33 6 6 43586 4 0 66
4. 920ms fim Standard When channel acquisition is in progress an acquisition indicator will display in the Channel Settings box The Image Show Hide button to the left of each Channel Settings box indicates the channel image data currently displayed In the example below Channel 1 image data is displayed and Channel 2 acquisition is in progress Please see Changing the Channel Image Display on page 51 for more information on viewing channel image data lt q gt Channel 1 Image Show Hide Buttons Channel Acquisition Indicator Image Information Selecting the Info box will display the initial image information By default the Title entry will be the date and time that the image was acquired During or after acquisition is complete the Title can be edited and the user project tags and notes for the image file can also be entered FluorChem E System User Guide Image Acquisition page 51 Info Edit 2011 01 1 3_15 18 00 Stop and Stop All Buttons The Stop button halts image acquisition only for the channel that is currently being acquired It will not halt acquisition for any subsequent channels If the protocol contains more than one channel the following prog ress bar and Stop All button will display Stopping exposure E StoP All Select the Stop All button to halt acquisition for all channels Otherwise acquisition will continue for the remaining channels When a protocol uses Auto expos
5. Add Channel Continue adding channels and changing settings as needed A total of up to four channels can be added Once a fourth channel is added the Add Channel button will no longer display FluorChem E System User Guide Overview of the Darkroom Screen page 47 Chemiluminescence j Channel 1 o p Channel 2 o pD Channel 3 oO Channel 4 op NOTE Channels that were added manually will be deleted automatically if a new protocol is selected in the Protocol Menu box Deleting Channels To delete a channel select the Channel Settings Menu box then select the Delete Channel button in the upper right corner vr Chemiluminescence X Channel 1 op Channel 2 o D Channel 3 x 4 Delete Channel Button No Light J No Filter gt b gt 7 5u Standard Channel 4 o D After a channel has been deleted the remaining channels will automatically renumber accordingly NOTE Imaging protocols must contain atleastone channel Channel 1 ina single channel protocol willnot be available for deletion FluorChem E System User Guide page 48 Chapter 5 Image Acquisition Image Acquisition The Acquire page will display in the Darkroom Screen once image acquisition has started Users can monitor and stop image acquisition view protocol and channel settings and channel image data in the Acquire page Darkroom Protocol lt 4 gt Channeli ImageDisplay Chann
6. Component Minimum Recommended Operating System Windows 7 Vista or XP Table 2 4 AlphaView software computer requirements when used on local workstations Networking the FluorChem E System The FluorChem E system can be connected directly to a local network for remote access using a web browser on any Mac or PC based network computer or Wi Fi enabled device such as a smartphone or iPad Image acquisition can be controlled and monitored remotely and stored image files can be accessed for copying printing and post acquisition desktop analysis using AlohaView software For more information on accessing the FluorCchem E system from a remote computer or device please refer to Chapter 9 Networking and Remote Access FluorChem E System User Guide System Installation page 11 System Installation The Fluorchem E system is easily installed by the user Once the space electrical and environmental require ments have been met 1 Unbox the system and place it on a lab bench IMPORTANT The FluorChem E system weighs 75 Ibs 34 kg Two people are required to lift the system from the shipping container onto the laboratory bench Lift the system by grasping the bottom of the instru ment at the midsection on either side Connect the power cord to the rear panel of the system and plug the cord into a power receptacle Network access optional To access Fluorchem E via a local network connect an Ethernet LAN cable to the 10
7. The top and bottom buttons double arrows will adjust the focus up or down in major increments Alternatively users can also enter a setting directly in the Focus Setting box Focus Position 3850 lt Focus Setting Box KP h proteinsimple Focus Adjustment Buttons y a Q j Expose Zoom Button Button As adjustments are made the value in the Focus Setting box will update automatically A new white light image will be taken and displayed each time the focus is adjusted Selecting the Expose button will take an additional epi white light image The Zoom button can be used to magnify white light images as desired When the desired focus is reached select Save to save the new setting The Focus Screen will close and the System Settings Screen will be displayed Select the Darkroom Tab A new white light image will be taken and displayed using the revised focus setting FluorChem E System User Guide page 102 Chapter 8 System Settings System Log To view or copy a Fluorchem E system log file select the Log button The following screen will be displayed View Log 03 01 13 30 54 999 INFO cellbio control imagingTask exposure time 10 0 bin 4 4 16 bit 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 03 01 13 31 07 346 IN
8. 100 0 32 9 73883 100 0 Channel 1 u 30 0 62470 7 34 47217 0 2 37 4 366166 33 6 lt Analysis 59 8 45556 4 0 48 9 32317 3 0 Define Lanes 35 0 474025 9 0 93 19454 37 E 32 1 21663 100 0 6 82 R3183 69 15 8 2285 6s 24 4 75950 6 3 343 176731 47 24 230260 19 7 676 148715 12 4 Boe 191877 16 0 94 2 200099 17 2 Ladder Lane 5 of 72 E oer tee Epa 4 Older Newer p Band Selection Individual bands can be selected and de selected by double clicking at the desired position on the image Bands can be added or removed from the analysis using the threshold slider or by clicking on individual bands For a multi channel image use the round channel buttons to limit analysis to a single channel and to select the bands to be analyzed To hide all the band indicators on the image select a channel and toggle the bands off Select All to analyze all the channels Move the threshold slider to raise or lower the intensity threshold for included bands The bands included are updated on the image as the slider is moved the table updates after the slider is released FluorChem E System User Guide Lane Profiling page 127 Adding and Removing Individual Bands To add a band to analysis 1 Move over the band on the image until the band indicator is shown 2 Click on the band To remove a band from analysis 1 Click on the band to highlight it in the table 2 Click again to remove the band FluorC
9. 32317 3 0 89 37722 3 5 ra 59 39135 7 3 25 474025 89 0 3 19454 3 7 5 44 21663 100 0 6 r 200 83183 6 9 s 150 22285 6 3 Too soso NEST 7 176731 14 7 7 236280 19 7 148718 12 4 j 191877 16 0 206699 17 2 FluorChem E System User Guide Exporting page 133 Exporting The analyzed image will reflect what is onscreen Band identifiers may be hidden or only shown for a single channel The tick marks may be percent or MW if entered in the table From the image Review results of the analysis can be exported to a computer in two ways foot AlphaView Open in AlphaView fcz files Images Images png files Results Results table csv file Cancel When exported as Images png files there will be two additional files included in the zipped folder that will have the following added to the file name _analyzed lanes The _analyzed png will consist of the image cropped to the region of interest and any identified bands and named lanes 207 40 60 80 100 The _lanes png will consist only of the cropped image FluorChem E System User Guide page 134 Chapter 11 Analysis in Digital Darkroom Printing The image printed will always reflect what is onscreen If analysis is enabled the printed image will be cropped to the region of interest To print the whole image analysis must be hidden FluorChem E System User Guide
10. User Guide Overview of the Darkroom Screen page 39 Protocol Channel Light Filter Sample Screen Chemi with Markers Nolight No filter 50 Epi White Orange 590 nm Table 5 1 Fluorchem E system pre programmed protocols The protocols available in the Protocol Menu depend on the filters installed in the FluorCchem E system Only protocols for the installed filters will be available for selection in the drop down menu Changing Channel Settings Light filter exposure and resolution settings for each channel used in a pre programmed protocol can be viewed and changed by selecting the Channel Settings Menu box These settings can be changed prior to acquiring an image NOTE Changes to default channel settings are not permanently saved When settings are modified the new selections are used for image acquisition until the Fluorchem E system door is opened At that time all channel settings will return to the default selections specified by the protocol FluorChem E System User Guide page 40 Chapter 5 Image Acquisition 7 Channel 1 No Light Light Settings F No Filter a __ Filter Settings O Auto a Exposure Settings su Standard a Resolution Settings Light Settings To change the light source select the Light Settings box and select the desired option Channel 1 No Light Ae O Epi White HUL Trans UV 4 5u Standard Filter Settings To change the
11. a web browser Stored image files can also be accessed remotely for reviewing copying printing and desktop image analysis using AlohaView software Connecting to FluorChem E Fluorchem E can be accessed remotely via a permanent URL address or a dynamic IP address NOTE The FluorChem E URL address is a unique and permanent address and will not change The IP address however isnot permanent and can change if the system is ever disconnected and reconnected to the local network FluorChem remote access via URL requires Apple Bonjour zero configuration networking to be installed on Windows based computers and devices Bonjour can be downloaded at http www apple com support bonjour Download and install Bonjour Print Services for Windows NOTES Once installed it is not necessary to launch Bonjour prior to accessing the Fluorchem E system remotely Bonjour capability comes pre installed on Mac iPad and iPhone and does not need to be installed sepa rately for remote web access of the Fluorchem E system To connect to a networked FluorChem E system 1 Open a web browser on a computer or device that is on the same network as the Fluorchem E system 2 Enter the URL or the current IP address for the FluorChem E system in the browser window The sys tem s URL and IP address information can be found in the System Settings Screen FluorChem E System User Guide Connecting to FluorChem E page 105 mm 1 6 FluorChem E FE023
12. aan bie 2 Introduction to the FluorChem E System 2 DOCU ENAS EIAS EE E A EE TAT 3 User Attention Notifications 000 00 ee 3 Perunin aroy La 0E vie ae rater vegans 3 Customer Service and Technical Support 5 OO OOM GG teat ght sco taipivat SEE aes 6 Fluorchem E System Disclaimer of Warranty 6 Chapter 2 INSTAN ATION ico scsi hha ba hon oenaretawiwk uae 7 MTOTO atts ins aae Ea kiana les 8 E exp OID IL ge a ae ne E eee een Sree 8 EECH OLR GUNT E os acd aa udtastnaes 8 Environmental Requirements 0 cece cece 10 Software and Computer Requirements 10 Networking the FluorChem E System 10 V HISTOMONON r T aban ete putts costar cuss 11 Chapter 3 System Hardware sssssssssssscsseee 13 Overview of FluorChem E System Components 14 PUOC RETENE UOTE anean anaE 14 STES TEE TE sa teste E aT 17 C e a E tah oenauesnatas 19 FluorChem E System User Guide page i EA T a E A E E Sesleeeres 19 UV Trais HUNG LON ig 2625 dices re oy BERR 20 General Guidelines and Information 20 System Operation Notification of Intended Use 20 Lifting and Moving the System Use Proper Lift MHO FEC COUO errre E e AEE NA 2al E E EE a E T EA adil 21 Chapter 4 Getting Starte sesiis rinni inie iiia 23 POWCTING ONT TES I e Ea 24 Overview of FluorChem E User Interface 29 USING the TOUSEN briand NAN 25 Uernedo Tee aia i 27 MPVOAGING SCR o E E E EE cits 31 SONI
13. cause permanent damage to the eyes and skin UV Interlock Switch The FluorChem E system is equipped with a safety interlock switch which automatically cuts off the power to the transilluminator when the door is open This eliminates any dangerous user exposure to UV light As a safety precaution the UV light will automatically turn off after three minutes when using the interlock switch To use the transilluminator for other purposes a key is required to override the safety interlock In this case be sure to use the following safety precautions Always wear UV protected eyewear specified by the manufacturer as providing protection at the wave length s used making sure the eyewear protects any areas where radiation may come through UV sunglasses may not prevent UV radiation from coming in through the sides or around the lenses Always cover all skin that may be exposed to UV light in particular the face neck hands and arms Always make sure that any UV protection devices such as the safety interlock switch on the transillu minator are working properly If not discontinue use until the device s are properly repaired Use only UV lamps in the transilluminator FluorChem E System User Guide Customer Service and Technical Support page 5 Hot Surface CAUTION Under normal conditions the temperature of the glass surface of UV transilluminator is safe to touch However if the system malfunctions it is possible that the gl
14. display in the Review Tab For more information on the Review Screen and making image adjustments please refer to Chapter 6 Image Review FluorChem E System User Guide page 78 Chapter 7 Managing Stored Images Searching for Images The Search box allows users to search for specific images in the Image Album To search for an image select the Search box A keyboard will display m la i A a KE Today EH EWV pE BR folie peg ie pe ee Gy pe AN ESA VB AEA DGA KEN PG PR Wi ai Yesterday shitt Z A Ga A B N M del 7 ES 7123 i a return SOI yL E v inga man m yy Aaa SSS SRT 1 LE FOLGE PS Ta 1e SOU eT _ E i ee l SoS Se 15 2 SRE ID eS POLE Leh ee LS Type in the text search information The following information can be used User entered information title user project and tags e Image file information date protocol exposure and resolution When text entry is complete select the Return button Search results will be returned in the Album page To clear the text in the Search box and return to a view of all files select the Clear button ch Chemi Clear Button FluorChem E System User Guide Editing the Image Album page 79 Editing the Image Album To delete or copy images in the Image Album select the Edit button This will display the Album Edit page The page will automatically scroll to the image currently displayed in the Review Screen and this image will be highlighted in
15. screen Chemiluminescent and epi white light imaging Non reflective and blocks residual UV light FluorChem E System User Guide page 18 Chapter 3 System Hardware e Clear screen UV imaging Transmits UV light e White screen Colorimetric gel imaging Converts transilluminating UV light to white light for imaging of visible stains Blue screen optional Converts transilluminating UV light to blue light for fluorescent imaging of blue excited stains An imaging area at the center of each screen assists users with sample positioning Prior to imaging samples should be placed on the appropriate sample screen and centered in the 16 cm x 12 cm imaging area The screen is then placed directly on the UV transilluminator proteinsimple Imaging Area When not in use sample screens should be stored in the screen holder provided with the FluorChem E sys tem FluorChem E System User Guide Compliance page 19 Compliance The FluorChem E system complies with UL 61010 1 2004 R10 08 Safety requirements for electrical equipment for measurement control and laboratory use Part 1 General requirements US EN 61010 1 2001 Safety requirements for electrical equipment for measurement control and labo ratory use Part 1 General requirements EU CAN CSA 22 2 No 61010 1 04 Safety requirements for electrical equipment for measurement control and laboratory use Part 1 General requirements CA EN 61326 1 2006
16. selected images Delete aticet Select the Delete button in the dialog box NOTE Images deleted on the Album Edit page are moved to the Trash page and are not permanently deleted FluorChem E System User Guide Trash Page page 81 Copying Images from the Album To copy one or multiple images from the Image Album select the image s in the Image List The copy function in the Album Edit page is the same as described in Copying Images on page 67 except that mul tiple folders are copied to the USB drive Exiting the Album Edit Page To exit the Album Edit page select the Cancel button Trash Page The Trash page contains images that have been deleted from the Review Screen and Album Edit pages Users can search and browse through all stored image files in the this page To view the Trash page select it from the Page Menu Page Edit Menu Button View Toggle Buttons gt sem Trash Edi earch J Search Box Today Last Week ii m en eee a y SOVIETS PDS Te SO IENE DONA See POE MATON AA BI ee EN PES PaS E en Fi E heal zi rs SOP D 28 12 3 7 DIG i A SOR ED GONA N D 2i aa HiT Two Weeks Ago T i 1 a SMALL POEL ATL AA aL T JON eA Trash page options are the same as the Album page Please refer to Overview of the Images Screen on page 74 for details on how to use this page Paging Buttons Image List Editing the Trash To delete copy or move image
17. the top of the list The Table view will automatically scroll to the image currently displayed in the Review Screen and this image will be highlighted in dark blue Paging Buttons The Paging buttons allow the user to page through data in the Image List The Page Up and Down but tons will scroll the Image List to the next or previous page of files The First and Last Page buttons will scroll the Image List to the first or last page of files NOTE When the Image List is at the first or last page of stored image files the Paging buttons will become inactive FluorChem E System User Guide Overview of the Images Screen page 77 Aa lt _ First Page a lt Page Up Ww lt Page Down wT Last Page Page Menu This menu allows users to switch between the Album and Trash pages All images are stored in the Album page by default When an image file is deleted in the Review Screen or from the Album Edit page it is moved to the Trash page To view image files in either page select the Page menu Album Album Trash Search Box The Search box allows users to enter text details to search for specific images Edit Button The Edit button displays the Edit pages where image files can be copied deleted or moved from the Trash page back to the Album page Selecting Images for Review or Enhancement To select an image for review or enhancement select a thumbnail or row in the Image List The selected image will then
18. 100 BASE T RJ 45 connector port Turn FluorChem E on using the power switch on the rear panel Let FluorChem E equilibrate for 30 minutes after the initial bower up before imaging samples This allows the system optics to reach the optimum acquisition temperature of 25 C FluorChem E System User Guide page 12 Chapter 2 Installation FluorChem E System User Guide page 13 Chapter 3 System Hardware Chapter Overview Overview of FluorCchem E System Components Compliance Safety Guidelines General Guidelines and Information System Specifications FluorChem E System User Guide page 14 Chapter 3 System Hardware Overview of FluorChem E System Components FluorChem Eis a high performance imaging system for Western blot and gel imaging and provides detection of chemiluminescent colorimetric and UV excited gels and blots The individual hardware components of the Fluorchem E system are described in this section FluorChem E Instrument Top USB Port LCD Touchscreen CCD Optics Motorized 4 Position Filter Wheel _ Epi White Lights Network and Side USB Ports System Door gt UV Transilluminator Door Latch UV Interlock Switch Integrated CCD Optics The FluorChem E system uses a scientific grade 8 3 megapixel CCD detector cooled to 25 C and a motor ized lens for optimum resolution and acquisition speed The optics are fully automated and require no manual user a
19. 14 Chapter 10 Maintenance and Troubleshooting 4 Next unscrew the four screws on the UV transilluminator inner front panel and remove the panel i 5 Slowly slide out the bulb tray NOTE Remove the bulb tray gently The bulb tray electrical cord is hard wired directly to the UV transillumi nator and the cord cannot be disconnected Pulling or tugging on the bulb tray may damage the cord and lead to malfunction in the UV transilluminator FluorChem E System User Guide Maintenance page 115 6 To remove a bulb rotate it until the pins align with the slot in the bulb socket Next lift the bulb up and out of the socket Bulb Tray Cord 7 Toinsert a bulb slide the pins into the bulb socket and then rotate the bulb a quarter turn A faint click will be heard when the bulb is seated correctly Middle Notch 8 Repeat steps 6 and 7 for the remaining five bulbs 9 Align the bulb tray on the middle notch of the UV transilluminator housing and slide the tray all the way back into the housing 10 Reinstall the inner front panel and tighten the four screws until snug 11 Reinstall the outer front panel and tighten the four screws until snug FluorChem E System User Guide page 116 Chapter 10 Maintenance and Troubleshooting 12 Push the UV transilluminator back into the system 13 Plug the FluorCchem E power cord back into the wall socket and turn the system power on Installing Emission Filters 1 Power
20. 2 bears Virila 220 Laboratory N FEO232 Mttp fe0232 local jiworh Network with DHCP 101239 lt lt ____ P Address 00 18 70 07 25 bs es nperature 25 0 c O Name Time Filters Update Test Dark Flat Focus Protocols Log Backup 3 The browser will expand the entered URL or IP address to http feXXXX local run_viewer browse_runs or http xx x x xxx run_viewer browse_runs and display the Images Screen mozilla Firefox File Edit View History Bookmarks Tools Help eo xX A E ete 0 1 1 185 run_viewer browse_runs r gt RE Geog ajl http 10 1 1 1 browse_runs ea Darkroom W Review E images ae Album Edit 5 Pal ti Today i gt a z eS eo DOUAR FEIDE JOLAIS ENa ee cm eee ee mee cer sD es ee Poh et oe ee ed eo Last Week a Bee Oe SO l a Aa gt alt 7 _ Boge ede Fr 307 too niai Two Weeks Ago a anpi 4 The full FluorChem E Digital Darkroom remote interface will now be accessible FluorChem E System User Guide page 106 Chapter 9 Networking and Remote Access NOTE If FluorChem E is executing a calibration or self test when accessed remotely an Instrument Test message will display in the Darkroom Tab If this occurs data can still be accessed from the Images and Review tabs Image acquisition and monitoring in the Darkroom Tab will remain unavailable until the calibration or test is complete Using the remote access interface is si
21. 3 06 3 38 13 678 INFO cellbio control imagingTask expose protocol Chemiluminescence id chemiluminescence 3 38 14 233 INFO cellbio control imagingTask exposure time 1 0 bin 4 4 16 bit 3 38 17 585 INFO cellbio control imagingTask save image dark_nofilter_1_1000 png 3 38 20 197 INFO cellbio control imagingTask exposure time 10 0 bin 4 4 16 bit 3 38 32 549 INFO cellbio control imagingTask save image dark_nofilter_1_10000 png 38 35 189 INFO cellbio control imagingTask exposure time 40 0 bin 4 4 16 bit 3 39 17 534 INFO cellbio control imagingTask save image dark_nofilter_1_40000 png 20 277 INFO cellbio control imagingTask exposure time 160 0 bin 4 4 16 bit 15 651 INFO cellbio control imagingTask camera stopped 16 309 INFO cellbio analysis correct corrections dark Use the scroll bar to review the log in its entirety To copy the log file insert a USB drive into any USB port on the instrument and select the Copy button FluorChem E System User Guide page 103 Chapter 9 Networking and Remote Access Chapter Overview Digital Darkroom Remote Access Connecting to FluorChem E Downloading Image Files Opening Image Files in AlohaView Software FluorChem E System User Guide page 104 Chapter 9 Networking and Remote Access Digital Darkroom Remote Access If Fluorchem E is connected to a local network image acquisition can be monitored and controlled directly from any networked computer or mobile Wi Fi device through
22. CS TE eronneen Bd SONDE POSITIONING esn iSS EENT AAA AET 31 OVE OVE O E os tented aE EOT 33 Chapter 5 Image Acquisition ssssssssssssess 35 Overview of the Darkroom Screen 006 006 36 WANG E A E I EEEE E PETE asta 36 CECOT O ar aaa 38 CHANGING CNGHNCLSCHINGS arisia Es ERRI 39 M lti Channel MAg N u wscrcocnen setters emadarts 45 page ii WIOGCACGU SION 2 tena lect a Aice cata ar 48 Changing the Channel Image Display 5 SOTIDICMING GINO ass hese EE EO wan ies 52 Chapter 6 Image RCVIEW oi cisco xcedes eirese rd as 55 Overview of the Review SCre n 00 cece cece ee 56 EE WMOGCINMONNGUON 42 622 Guna te westat tune 59 Channel Image Display and Overlay 60 Viewing Image Data for One Channel 60 Overlaying Channel Image Data 6 Making Image Adjustments 6 06 0000 62 Bron TES CONTON os tx suri ece ceive na Eaei 62 LULL AIETEN TEE A ETE SEE ATAOI EY 63 EXO OSUICS VICW peau kd waGu Rie Gentose sangeet cuss 63 WINGO C ZOOM uer a a EaR 64 VICWHEG PIXE VI a a riran aa 66 DOW NG HINA CS aranera N E S 6 Fluorchem E System File Information 67 Copying Printing and Deleting Images 67 COPAG ITO E eari siira EET 6 PRURIT OJE wate kd iba the Gena ln scm gdh duke 69 DECI INO CS rn wneud e ee ketene 71 Chapter 7 Managing Stored Images 73 Overview of the Images SCIEN 6 cece cece 74 Vew rog die DONON ror enna T 74 MAIE
23. EST nE E EAA 74 POGIIG DUNO aa arIa TAR 6 POG SINC 6 ta adhere Ranken Gabe ais eens a ad Lr DECC OOK ats eco E E E ET IF POM PUNON At e E O 77 FluorChem E System User Guide Selecting Images for Review or Enhancement 77 ETIT N INOG CS Are Gees 78 FOMING tielnage ADUN kire ronin REEI 79 Selecting Images in the Album Edit Page 79 Deleting Images from the Album 80 Copying Images from the Album 81 EXIUNG ANE AIOUN ECIT POG E ah eayitsceitnenats 8 WRG AOC AE NEEE E E ETTE OEA 8 ENING CCN VAS Mik dentate a tiilcas SEA 8 BOCKUDIGNG RESTON cui tha rates autide teens i 83 E E i EE Ce ee ee E ee eee eee 83 OSMO NCES to hhi et a san Sha AeA ase 85 Chapter 8 System SettingS cc cceccecccccecaes 87 Overview of the System Settings Screen 88 WEMA 42 ectctunacn naigit lt kiden 88 Software and Network Information 89 OPUS CID CTOL ONC e ss AEE CaA NETR 89 Changing SYSTEM INIOHNGUON eberr tiia 89 Changing Date and Time Settings erreren i 90 Changing Her aG e amp csattntn ar aa 9 WOOGUNG Cie SONW AIC sires cits eae euead de 93 DSB OO GICS erirken i ebakiera e leie dekket 93 Wea TE ee a a 93 DOVING UOT OOTO a aE EEE 95 VS e T aeiae R 96 RUNNING Ne sel VOSS rirerire ansni 97 Creating DAK MOSTC S suena a nea saan dele 97 FOER EGC ANNGO o r E EE ied ey Waist 98 CRONGING r ocuUs S eT erea EE 100 aa E S 102 Chapter 9 Networking and Remote Access 103 Digital Darkroom Re
24. Edit Today s IMMM IM 7 z E z AG HL RR TS BONA A SGT ES RSET eed FOTN a BST 1b Pa ee Se ee ot er Last Week l FULTS are Tie ie ee ee eT ee ee hm ee ee ST Two Weeks Ago BOIERI Teed PST EET BS TEET FluorChem E System User Guide i a y T k Tie ee ET E TIET N Pi F VW page 30 Chapter 4 Getting Started System Settings Screen Selecting the tab on the far right will display the System Settings Screen This screen displays system information and allows users to change the system name date and time and focus settings as well as install software updates and run system tests Dark masters flat field calibration and system log files can also be gen erated Darkroom FluorChem E FE0232 eae Laboratory ote ae iN FEO2 32 http fe0232 local Network with OHCP 101239 DO 18 7d Of 25 bs ot Temperature 25 0 C CS Name Time Filters Update Test Dark Flat Focus Protocols Log Backup Screen Saver After a period of inactivity the FluorChem E User Interface will automatically launch a screen saver to pre serve the lifetime of the LCD touchscreen When the screen saver is running the screen will be blank Simply touch the screen to exit the screen saver mode and view the user interface FluorChem E System User Guide Imaging Setup page 31 Imaging Setup Prior to image acquisition samples must be placed on the appropriate sample screen and positioned pr
25. Electrical equipment for measurement control and laboratory use EMC Require ments General requirements EU Safety Guidelines IWARNING If the instrument is not used as specified by the manufacturer the overall safety will be impaired WARNING If the system is damaged and does not function properly stop the instrument safely and contact Pro teinSimple Technical Support immediately CAUTION Avoid using the FluorChem E system in a manner not specified by ProteinSimple While the system has been designed to protect the user this protection may be impaired ifthe instrument is used improperly FluorChem E System User Guide page 20 Chapter 3 System Hardware UV Transilluminator CAUTION The FluorChem E system contains an integrated ultraviolet transilluminator The system enclosure confines the radiation within the system and shields the user from exposure Exposure to UV radiation can cause permanent damage to the eyes and skin CAUTION Under normal conditions the temperature of the glass surface of UV transilluminator is safe to touch However if the system malfunctions it is possible that the glass surface tempera ture could exceed 80 C Please exercise caution when touching the glass surface UV Interlock Switch The FluorChem E system is equipped with a defeatable safety interlock switch which automatically cuts off the power to the transilluminator when the door is open This eliminates any dangerous us
26. FO cellbio control imagingTask save image dark_nofilter_1_10000 png 08 734 INFO cellbio analysis correct corrections dark 11 237 INFO cellbio control imagingTask exposure time 0 2 bin 4 4 16 bit 13 784 INFO cellbio control imagingTask save image dark_nofilter_2_200 png 15 221 INFO cellbio analysis corect corrections dark 17 749 INFO cellbio control imagingTask exposure time 0 3 bin 4 4 16 bit 20 401 INFO cellbio control imagingTask save image dark_nofilter_3_300 png 21 904 INFO cellbio analysis corect corrections dark 24 003 INFO cellbio control imagingTask save run FCE_0232_2011 03 01_13 30 54 3 33 06 536 INFO cellbio control imagingTask expose protocol Chemiluminescence id chemiluminescence 33 07 093 INFO cellbio control imagingTask exposure time 60 0 bin 4 4 16 bit 3 34 09 452 INFO cellbio control imagingTlask save image dark_nofilter_1_60000_png 3 34 10 983 INFO cellbio analysis corect corrections dark 34 13 478 INFO cellbio control imagingTask exposure time 30 0 bin 4 4 16 bit 34 45 828 INFO cellbio control imagingTask save image dark_nofilter_2_30000 png 3 34 47 133 INFO cellbio analysis corect corrections dark 3 34 49 633 INFO cellbio control imagingTask exposure time 30 0 bin 4 4 16 bit 3 35 21 978 INFO cellbio control imagingTask save image dark_nofilter_3_30000 png 3 35 23 269 INFO cellbio analysis correct corrections dark 35 25 359 INFO cellbio control imagingTask save run FCE_0232_2011 03 01_13 3
27. Installation FluorChem E System User Guide page 8 Chapter 2 Installation Introduction This chapter will help prepare the laboratory for installation of the ProteinSimple FluorChem E system Please complete the space electrical and environmental requirements prior to installing the instrument Space Requirements The FluorChem E system is a bench top instrument which requires a stable laboratory bench or table capable of supporting 75 Ibs 34 kg System dimensions and space requirements are as follows Dimension Centimeters Inches Table 2 1 FluorChem E system dimensions A minimum clearance of 3 inches 7 6 cm at the rear of the instrument must be provided for the CCD cool ing fan ventilation and sufficient access to connect or disconnect the power cord as well as to operate the power switch Electrical Requirements As with all electrical instruments the FluorChem E system should be located away from water solvents or corrosive materials The system should also be placed away from interfering electrical signals and magnetic fields Installation of the FluorChem E system requires a dedicated grounded circuit capable of delivering the appro priate current and voltage for your country The power requirements for select countries are listed in Table 2 2 FluorChem E System User Guide Electrical Requirements page 9 Frequency Region Volts AC Hz Amps oma a o te Table 2 2 FluorChem E system power requ
28. Tiff File 6 26 2010 3 19 PM E Documents a Music E Pictures E Videos Computer File name BEE Save as type Tif Fies e80 o dade FluorChem E System User Guide page 110 Chapter 9 Networking and Remote Access Opening png Files To open saved or extracted png files on the remote computer open AlphaView software Select File gt Open Browse to the file location and select the desired png file NOTE FluorChem E png image files can also be opened in any other program that accepts files in png format FluorChem E System User Guide page 111 Chapter 10 Maintenance and Troubleshooting Chapter Overview Maintenance e Accessories and Spare Parts Troubleshooting FluorChem E System User Guide page 112 Chapter 10 Maintenance and Troubleshooting Maintenance Cleaning the FluorChem E System Instrument and Sample Screens Items needed Cleaning solution general purpose glass cleaner isopropanol or 70 ethanol and a soft cloth or paper towels 1 Saturate a paper towel or cloth with cleaning solution 2 Wipe down the surface of the UV transilluminator sample screens and the instrument s exterior blue housing LCD Touchscreen Items needed LCD screen cleaner Klear or iKlear and a microfiber cloth 1 Follow the instructions provided on the LCD screen cleaner bottle NOTE Never spray cleaning solution directly onto the LCD touchscreen 2 If there are no instructions on the cleaner b
29. ancel button Backup and Restore Backup Acquired images are stored on the internal hard drive of the imager It is recommended that images be backed up regularly to an external USB flash drive NOTE Backup and Restore is not visible on the remote browser page only at the instrument To initiate the backup select Backup on the Settings screen W Review fS Images FluorChem E FEO232 oftware Version 3 0 0 Laborato ou Feozgz MTP fenia otal Dvi States Metroak wiih HEP rr 18 of 3 a op T mperiluii e Z230 C CG Name Time Filters Update Test Dark Flat Focus Protocols Log Backup ___ Backup FluorChem E System User Guide page 84 Chapter 7 Managing Stored Images After selecting Backup it will open a dialog giving you the choice to initiate a Backup or Restore Restor ing images from a USB will not overwrite existing files on the imager Backup and Restore x Backup Save all images to a USB drive Restore Restore images from a USB Drive Cancel The following dialog box appears if no USB is detected Backup and Restore No USB drive detected Insert drive and select Retry Retry Cancel Backup displays a dialog box containing info on the backup before it starts Backup and Restore Detected USB Drive This backup will contain 10 images This may take several minutes Start Cancel If the USB drive does not have room for the backup a warning is given wit
30. annel image capture on up to four channels per acquisition when needed for specific applications Each channel can then be subsequently overlaid or stacked in the Review Screen for visualization of images such as e Colorimetric markers on chemiluminescent blots e Chemiluminescent blots at various exposure times Adding Channels Once a protocol is selected a total of up to four image capture channels can be added prior to acquisition A common application requiring additional channels would be when multiple manual chemiluminescent expo sures are desired All protocols must contain at least one channel The Channel Settings Menu box will display icons repre senting light and filter settings used for each channel These icons will automatically update whenever channel settings are changed To add a channel select the Add Channel button bd Chemiluminescence i gt Channel 1 Add Channel Add Channel Fa Button The new channel added will by default display the same settings as the previous channel FluorChem E System User Guide page 46 Chapter 5 Image Acquisition 2 Chemiluminescence z gt Channel 1 ey Channel 2 NoLight P No Filter LEF 5u Standard Add Channel Change light filter exposure and resolution settings as desired 3 Chemiluminescence gt Channel 1 op Channel 2 Noligh p No Filter gt VF Manual Time 20 0 seconds 5u Standard
31. are FluorChem E System User Guide Safety page 3 Safety User Attention Notifications Several user attention phrases are used throughout this manual Each phrase should draw the following level of attention from the user NOTE Points out useful information IMPORTANT Indicates information necessary for proper instrument operation CAUTION Cautions users regarding potentially hazardous situations in regard to user injury or dam age to the instrument if the information is not heeded WARNING Warns users that serious physical injury can result if warning precautions are not heeded Instrument Safety Labels The following safety labels are located on the FluorChem E system Each label will display a safety alert symbol indicating a potential safety hazard Symbol Description Refer to the FluorChem E User Guide before proceeding Table 1 1 Fluorchem E system safety labels FluorChem E System User Guide page 4 Chapter 1 General Information WARNING If the system is not used as specified by ProteinSimple overall safety will be impaired WARNING If the system is damaged and does not function properly stop the instrument safely and contact Pro teinSimple Technical Support immediately UV Safety CAUTION The FluorChem E system contains an integrated ultraviolet transilluminator The system enclosure confines the radiation within the system and shields the user from exposure Exposure to UV radiation can
32. ass surface tempera ture could exceed 80 C Please exercise caution when touching the glass surface Electrical Safety Be sure to take proper precautions when handling any electrical equipment Never work on any live circuit fix ture receptacle or switch Safety rules to follow whenever working with any electrical appliance include Always turn the system power off and unplug the power cord before changing a fuse Always shut off power to the circuit before repairing or replacing a switch receptacle or fixture Always tape over the main switch empty fuse socket or circuit breaker being worked on Always check that the circuit is dead before beginning work on it Using a circuit tester or voltmeter can help determine this Always unplug any appliance before repairing it Prior to System Operation Ensure that all users of the FluorChem E system have Received instruction in general safety practices for laboratories Received instruction in specific safety practices for the instrument Received instruction on handling of biohazards if biohazardous materials are to be used on the system CAUTION Avoid using the Fluorchem E system in a manner not specified by ProteinSimple While the system has been designed to protect the user this protection may be impaired ifthe instrument is used improperly Customer Service and Technical Support The ProteinSimple support staff is available to respond to any questions or comments about our p
33. box will display Delete this image Select the Delete button in the dialog box NOTE Images deleted from the Review Screen are moved to the Trash page and are not permanently deleted After an image has been deleted the Images Screen will display For details please refer to Trash Page on page 81 FluorChem E System User Guide page 72 Chapter 6 Image Review FluorChem E System User Guide Chapter 7 Managing Stored Images Chapter Overview Overview of the Images Screen Editing the Image Album Trash Page FluorChem E System User Guide page 73 page 74 Chapter 7 Managing Stored Images Overview of the Images Screen Selecting the Images Tab will display the Album page of the Images Screen This screen is used to browse saved images Page Edit Menu Button Darkroom View Toggle Buttons gt Album Edit ol Search Box o Image Title JOINED tappie Siano Iaia POOROTIERLSG OnO TEMES SONTORSTVE bal 101103At ILII SOltaeOT abcd w Paging Buttons Image List Last Week E ee ra ZOU SSSALTD4SS 20110224 102008 2011 0228 434 20110222100529 2011023 Two Weeks Ago SOUT SSP LIS4S4 BOTISDICOS4E1E 2011 0217 104627 ME rT GIST JOTiLO22ILI620 2Nar 202 View Toggle Buttons The View Toggle buttons allow users to change how the list of image files is displayed The view can be tog gled between viewing image files as Thumbnails or list
34. cquire page will display and Channel 1 acquisi tion will begin FluorChem E System User Guide page 54 Chapter 5 Image Acquisition Darkroom Chemiluminescence lt 4 gt Channeli Channel 2 o Channel 3 e gt Info Channel 1 1 00m exposure Taking 2 40m exposure M Stop For channels using Auto expose image acquisition will continue automatically until the correct exposure is determined Ifthe channel is using Manual expose image acquisition will halt when the exposure time entered in the channel settings is reached Image acquisition can be halted at any time by selecting the Stop button NOTES Multi channel acquisition willoccur sequentially Selecting the Stop button during amulti channelacquisi tion halts acquisition only for the channel being acquired It will not halt acquisition for any subsequent channels To halt acquisition on all channels select the Stop All button that becomes available after Stop is selected Image data will only be available for those channels that completed acquisition When the Chemi with Markers protocol is used for image acquisition the chemiluminescent image of the sample Channel 1 will display in grayscale and the white light image of the markers Channel 2 is dis played in red to optimize visualization When acquisition is complete all channel data is processed and final image s will be displayed in the Review Tab which will be described in deta
35. d Technical Support Legal Notices FluorChem E System User Guide page 2 Chapter 1 General Information Welcome Congratulations on your purchase of the FluorCchem E system ProteinSimple welcomes you as a new user This user guide includes important safety information installation requirements maintenance procedures descriptions of the instrument and user interface as well as detailed operating instructions for setting up and running FluorChem E Introduction to the FluorChem E System FluorChem E is a high performance imaging system for quantitative Western blot detection and gel imaging Its compact design incorporates state of the art CCD optical technology with an intuitive touchscreen inter face for detection of chemiluminescent UV excited and colorimetric gels and blots In addition users can con troland monitor image acquisition and access data remotely using a unique browser based interface from any networked computer or mobile Wi Fi device The FluorChem E system comes with a cooled 8 3 megapixel CCD detector 50 mm f 1 4 motorized fixed lens 4 position motorized filter wheel and 590 50 nm orange emission filter Light sources include epi white lights trans white light via white light conversion screen and an integrated 302 nm UV transilluminator The system also includes an on board integrated computer and LCD touchscreen the FluorChem E Digital Dark room User Interface and AlphaView stand alone image analysis softw
36. d with the current protocol and or added by the user Users have the option of adding channels by select ing the Add Channel button Selecting the Channel Show Hide button again will hide the channel display For more information on channels please see Multi Channel Imaging on page 45 Channel Show A z Hide Button gt Chemiluminescence ig Chemiluminescence 7 t Channel 1 Add Channel Channel Settings Menu The Channel Settings Menu box allows users to view and change light filter exposure and resolution set tings for each channel These settings can be viewed by selecting the Channel Settings Menu box FluorChem E System User Guide page 38 Chapter 5 Image Acquisition i Chemiluminescence 7 Close Settings View gt Channel 1 NoLight 7 No Filter O Auto v 5u Standard Add Channel The Channel Settings Menu box displays icons representing the light and filter settings used for each channel These icons will automatically update whenever channel settings are changed To close the settings view select the orange arrow or channel number in the upper left of the Channel Set tings Menu box Expose Button The Expose button starts image acquisition Selecting a Protocol Protocol selection depends on the sample being imaged Table 5 1shows a list of pre programmed protocols along with the light setting filter setting and the recommended sample screen for each FluorChem E System
37. dark blue Delete Copy Button Button i mages or E an Button Select All Button Select All Paging Buttons Last Week oo 4 E UNRATE F e E rt E a Fed E a E Ei eaa a a E a 1 EO Woke VET A OTTAA TAA BONNET VET DEP IE k 2 Image List Two Weeks Ago Wl PE ee ee ee ee eee Selecting Images in the Album Edit Page One or multiple images can be selected in the Album Edit page for subsequent deletion or copy To select images select the thumbnail s or row s of the desired images in the Image List Once images are selected the Delete and Copy buttons will become active Selected images will be designated with a check mark and the number of images selected will display in the upper left corner of the page FluorChem E System User Guide page 80 Chapter 7 Managing Stored Images 3 Images Selected f Delete 2 Cony Today Oe Select ES sa hes E EI EI f l J301 Ba 0301 161620 2077 Jaia Ewes 1 02 01 1 T r ER we Last Week ad o E JE z 122 23 11 83 01 2071 02 pf MALOGA HUAN IENA MSANI F Two Weeks Ago 1a Gaade ae The Select All button can be used to select all images on the current page Selecting the button again dese lects all images on the page Deleting Images from the Album To delete one or multiple images from the Image Album select the image s in the Image List then select the Delete button The following dialog box will display Delete
38. de FluorChem E System User Guide page 24 Chapter 4 Getting Started Powering on the System To power up the FluorChem E system turn on the main power switch located at the rear of the instrument NOTE When imaging chemiluminescent samples allow the FluorChem E system to equilibrate for at least 30 minutes after power up This ensures that the system optics have reached the optimum acquisition temperature prior to imaging This equilibration time is not necessary if the system power remains on at all times When the system has finished booting up and is ready to use the following screen will display Darkroom Colony Plates Chemiluminesence UV Colorimetric Center blot on black screen Center gel on clear screen Center sample on white screen Place screen on light box then close door i FluorChem E FE0232 Laboratory http fe0232 local Ce Be NOTES ProteinSimple recommends leaving the Fluorchem E system power on when not in use Power the system down only when it will not be used for extended periods of time Cycling the system power on a regular basis is not optimal for the system s integrated computer and cooled CCD optics The FluorChem E system must be powered on in order for a remote computer or device to connect and access the browser version of the FluorChem E User interface through a local network connection FluorChem E System User Guide Overview of FluorChem E User Interface page 25 Ove
39. djustments FluorChem E System User Guide Overview of FluorChem E System Components page 15 LCD Touchscreen The 10 4 LCD touchscreen is used for direct control of the instrument through the FluorChem E User Inter face Image acquisition and review system settings and image file copy and printing options are accessed through commands on the touchscreen Simply touch the screen to see menus or activate controls Protein Simple recommends using the stylus provided with the system to interact with the touchscreen System Door The FluorChem E system door provides access to the inside of the instrument for loading of samples gels and blots for imaging To open the door pull the door latch forward and down until it stops then pull the door open To close and lock the door close the door and turn the door latch up and back to its stop position NOTE The FluorChem E system door must be closed during image capture for proper instrument opera tion and user safety Epi White Lights Epi white lights are also located at the top of the inside cabinet The white light produced is used for sample positioning and imaging of colorimetric blots UV Transilluminator An integrated UV transilluminator is located at the base of the inside cabinet and is used for excitation of stained DNA and protein gels The unit slides out for easy sample loading Power to the UV transilluminator is automatically disabled when the FluorChem E system door is op
40. down the FluorChem E system 2 Open the system door and locate the panel in front of the filter wheel at the rear of the inside cabinet Panel Screws 3 Using a Philips head screwdriver unscrew the three screws on the base of the panel and remove the panel This will expose the filter wheel Each filter position is labeled with a position number NOTE Filter Position 1 is reserved for chemiluminescent imaging and should always remain empty FluorChem E System User Guide Maintenance page 117 4 To installa filter insert the threaded side into an open port on the filter wheel Turn clockwise to screw the filter in place until snug To remove a filter grasp the outer edge and turn counter clockwise to unscrew and remove When filter installation is complete replace the panel Turn the FluorChem E system power on 0 N 2 pi Update the Filter Settings in the System Settings Screen For details on how do to this please refer to Changing Filter Settings on page 91 FluorChem E System User Guide page 118 Chapter 10 Maintenance and Troubleshooting Changing the Fuse 1 Power down the FluorChem E system and unplug the power cord from the system 2 Locate the fuse holder on the rear panel of the instrument Fuse Holder Use a flat head screwdriver to gently pry the fuse holder open Remove the fuse holder Remove both fuses and replace them with two 250 V 6 A fuses P N 011 597 Reinsert the fuse hold
41. e selecting the Stop button will halt image acquisition and save the last completed exposure as the final sample image When a protocol uses Manual expose selecting the Stop button will halt image acquisition and discard any exposure data Changing the Channel Image Display During acquisition the Image Display will update to show image data for each channel as it is acquired However image data for any channel can be displayed by selecting the Image Show Hide button next to the Channel Settings box The button can be toggled between Show arrow only and Hide red line through arrow i o Image Hide a adh acak i Fl Channel Image Show _ Channel3 FluorChem E System User Guide page 52 Chapter 5 Image Acquisition NOTE Image data for only one channel at a time can be displayed during the acquisition process To view image data for a different channel toggle the Image Show Hide button next to the desired Chan nel Settings box to Show The image data for the selected channel will be shown and all other channel image data will be hidden The Image Show Hide button s for the remaining channels will change to Hide automatically Sample Imaging NOTES When imaging chemiluminescent samples allow the FluorChem E system to equilibrate for at least 30 minutes after power up This ensures that the system optics have reached the optimum acquisition tem perature prior to imaging This equilibration time is not
42. e changed Image adjustments only change the way the image is visualized They do not alter the original image data or affect quantitative results Brightness Controls The Brightness Controls allow users to make images lighter or darker via a non linear gamma adjustment To adjust image brightness select the up or down arrows The setting control indicator will increase or decrease as adjustments are made All images have a default setting of zero FluorChem E System User Guide Making Image Adjustments page 63 Invert The Invert function inverts the gray levels of the displayed image converting a positive image to fs negative or vice versa For example an image with black bands on a white background will be con verted to an image with white bands on a black background Selecting the button a second time returns the image to its original form NOTE Image adjustments only change the way the image is visualized They do not alter the original image data or affect quantitative results Bands in the image will have the same quantitative densities in both the original and inverted versions of the image Chemiluminescent Image Chemiluminescent Image Before Invert After Invert Exposures View The Exposures View feature in the Review Screen displays additional exposures of the acquired image and allows users to view and select from nine different exposure levels Once the Expo sures View button is selected the follow
43. e number of lanes into the text box or by using the and buttons to increase or decrease the number of lanes within the region of interest Number of lanes 6 Number of lanes Rotate UV Fluorescence a Region of Interest FluorChem E System User Guide page 124 Chapter 11 Analysis in Digital Darkroom Defining Region of Interest Define the region of interest by moving and resizing the rectangle overlaid on the gel or blot image Select Analyze to continue or Cancel to leave analysis Number of lanes 6 Rotate T T UV Fluorescence Region of Interest FluorChem E System User Guide Lane Profiling page 125 Rotating Region of Interest The region of interest can be adjusted for rotation using the two Rotate buttons Clicking on these buttons will rotate the image 1 in a clockwise or counterclockwise direction with the center of rotation defined by the rectangular overlay Rotate Number of lanes 6 Tr Rotate S UV Fluorescence a gq FluorChem E System User Guide page 126 Chapter 11 Analysis in Digital Darkroom Calculating Intensity Values Based on the region of interest and number of bands Digital Darkroom will automatically crop and zoom in on the area of interest locate the bands in each lane and calculate the intensity Darkroom W Review E Images Coomassie Blue P Info t 0 Lane Position letemsity NTotal Adjust f 33 3 166738
44. e setting in the Channel Settings box The Image Show Hide button to the left of each Channel Settings box indicates what channel image data is currently displayed In the example shown next image data for Channels 1 and 3 are shown and Channel 2 image data is hidden Please see Channel Image Display and Overlay on page 60 for more infor mation on viewing and overlaying channel image data lt Channell Image Show Hide Buttons Ao gt Channel2 lt gt Channel3 Image Information Any sample and image information entered by the user is also stored with all images This information can be viewed by selecting the Info box Info Edit i 2011 01 19_15 18 00 s Dai NOTE If the Auto Expose setting was used and image acquisition was stopped by the user Auto Stopped see protocol details will display as the title in the Info box FluorChem E System User Guide Entering Image Information page 59 Image Review Options The buttons located under the Image Display allows users to browse between files currently displayed on the Images Tab as well as Delete Copy and Print images In addition the Zoom feature provides image magnification 0 Delete eked Oe a tat Print 4 Older Newer p Q Delete Copy Print Browse Images Zoom Entering Image Information To enter sample image information select the Info box Then select the Edit button The Image Info page will display mage info EE
45. ed in a Table View View EEE Thumbnails Daca Table Image List The Image List displays a complete list of all image files stored on the FluorChem E system The Image List display depends on the View Toggle button selected FluorChem E System User Guide Overview of the Images Screen page 75 Thumbnail View In the Thumbnail view of the Image List file thumbnails are shown for each image Image files are sorted by date and time with the most recent files at the top of the list Image titles are listed below each thumbnail The Thumbnail view will automatically scroll to the image currently displayed in the Review Screen and this image will be highlighted in dark blue NOTE If the Auto Expose setting was used and image acquisition was stopped by the user Auto Stopped see protocol details will display as the image title Today 207 1 03 01_16 2 2077 03 01_15 18 25 2077 03 07_ 6 26 29 2071 03 01_15 16 56 2071 03 01_13 48 4 2011 03 01_13 38 13 2011 03 01_13 33 06 MRREtre ees eT 2011 03 01_12 08 48 2017 03 01_12 02 50 Last Week EERE gt 2011 02 24_12 40 57 2011 02 24_10 20 08 2011 02 23_11 4 25 2017 02 21_16 29 18 2017 02 21 1 1 Two Weeks Ago n 2011 02 17_13 48 41 2011 02 18_02 48 16 2071 02 17_10 46 27 Table View In the Table view of the Image List image files are shown in a table format Table columns include User title project and tags entered by the user e Acquisition protoc
46. een updated select the Save button to save the changes Please refer to Installing Emission Filters on page 116 for instructions on installing filters in the filter wheel FluorChem E System User Guide Updating the Software page 93 Updating the Software Software updates can be transferred to the FluorChem E system using a USB flash drive or downloaded via the web USB Updates 1 Insert the USB drive into any USB port on the instrument 2 Select the Update button The following dialog box will display Software Update Check for software updates on the web JSE di USE Drive Install software from a USB drive 3 Select the USB Drive button 4 The software will perform an automatic scan of the USB drive to locate the software installer file When prompted follow the on screen instructions to complete the software update Web Updates NOTE Web updates require that the FluorChem E system be connected to a local network with internet access 1 Select the Update button The following dialog box will display FluorChem E System User Guide page 94 Chapter 8 System Settings Software Update x Web Update Check for software updates on the web a saaa Install software from a USB drive 2 Select the Web Update button 3 The software will automatically check to see if a new version of the software is available If one is found follow the on screen instructions to comp
47. einSimple recommends starting the process in the evening and letting it run overnight 1 Close the FluorChem E system door 2 Goto the System Settings Tab and select the Dark button The following dialog box will display Create Dark Masters Dark Masters created 04 Jan 2011 This will take multiple long exposures with the camera shutter closed Estimated duration 12 01 hours D is ia BF Wwerter i a 1 E 4 gt a gt FluorChem E System User Guide page 98 Chapter 8 System Settings i Select Start This following status screen will display gt Create Dark Masters Taking dark frame exposures Started 09 03 AM 15 Feb 2011 Image 1 152 Exposure 50ms _ Expected completion 09 04 PM 15 Feb 2011 The start and expected completion times will be shown along with an image counter and progress bar which will update during the Dark Master file generation process NOTE To maintain optimal light tightconditions avoid opening the FluorChem E system doorafter starting Dark Master file generation When the Dark Master files have been created the following message will display to indicate the process is complete Create Dark Masters x Dark Masters created 15 Feb 2011 Click OK to exit Flat Field Calibration Flat field calibration is used to correct for optical non uniformity inherent in CCD based imaging systems This calibration is applied automatically as par
48. el tp AABN S nD Information Channel 3 Image gt Info Information Channel 1 1 00m exposure Stop Prog ress Bar gt Taking 2 40m exposure _ _ a Stop LO Button Image Display The Image Display area will display the image as it is acquired The channel number and exposure time will be displayed under the Image Display When a channel uses Auto expose the Image Display and exposure time will refresh each time a new exposure is taken during image acquisition FluorChem E System User Guide Image Acquisition page 49 Channel 2 150ms exposure Taking 10s exposure iam When a channel uses Manual expose the Image Display and exposure time will not present data until the image has been taken at set exposure time The acquisition indicator will display while imaging is in progress Sa Taking 100ms exposure gS Progress Bar The Progress Bar will indicate the FluorChem E system status during image acquisition and will refresh as each exposure is taken FluorChem E System User Guide page 50 Chapter 5 Image Acquisition Protocol Information The Protocol box will display the protocol currently being used for image acquisition Channel Information The Channel Settings box es will display the individual channel settings used for image acquisition To view light filter exposure and resolution settings select the Channel Settings box lt Channel 1 UUI Trans UV Orange O Auto Time
49. em door is closed The sample is now ready to be imaged FluorChem E System User Guide UV Light Override page 33 UV Light Override If necessary users can override the safety interlock switch and turn on the UV transilluminator manually To do this 1 Open the system door The Door Open Screen will display Colony Plates Colorimetric Chemiluminesence Center blot on black screen Center gel on clear screen Center sample on white screen Place screen on light box then close door FluorChem E FE0232 Laboratory http fe0232 local pay GV Eight 2 Select the UV Light button on the touchscreen The following UV light dialog box will display UV Light N Override the safety switch to turn on the UV light Pull Style Push Styte i mun t Caution UV Light Always wear eye protection 3 Insert key and turn clockwise FluorChem E System User Guide page 34 Chapter 4 Getting Started NOTE As a safety precaution the UV light will automatically turn off after three minutes when using the interlock switch CAUTION Exposure to UV radiation can cause permanent damage to the eyes and skin Always wear UV pro tected eyewear and protective clothing over areas of skin that may be exposed to UV light when engaging the UV interlock switch override 4 When finished turn off the UV transilluminator by turning the key couterclockwise and removing Then select the Off button to exit the UV light dialo
50. en CAUTION The Fluorchem E system contains an integrated ultraviolet transilluminator The system enclosure confines the radiation within the system and shields the user from exposure Exposure to UV radiation can cause permanent damage to the eyes and skin The FluorChem E system UV transilluminator uses 302 nm bulbs These bulbs can be replaced with 365 nm bulbs if needed Please refer to UV Transilluminator Bulb Replacement on page 113 for information on replacing UV transilluminator bulbs FluorChem E System User Guide page 16 Chapter 3 System Hardware UV Interlock Switch A defeatable safety interlock switch located below the UV transilluminator automatically cuts off power to the transilluminator when the door is open This eliminates any dangerous user exposure to UV light As a safety precaution the UV light will automatically turn off after three minutes when using the interlock switch When the system door is open users can override the safety interlock switch and turn the UV light source on by inserting the supplied hardware key and turning clockwise The switch can only be used when the UV Light button has been selected from the Door Open Screen in the user interface For more information on how to use this option please refer to UV Light Override on page 33 CAUTION Exposure to UV radiation can cause permanent damage to the eyes and skin Always wear UV protected eyewear and protective clothing over areas o
51. er SY oi gS W Plug the FluorChem E power cord back into the system and turn the power on FluorChem E System User Guide Accessories and Spare Parts Accessories and Spare Parts page 119 A set of common spare parts accessories and service information is listed below To place an order please call 888 607 9692 Part Description Part Number Accessories Chemiluminescence Imaging Screen Black 90 15045 00 UV Gel Imaging Screen Clear 90 15044 00 White Light Conversion Screen White 90 15042 00 Blue Light Conversion Screen Blue 90 15043 00 Screen Holder 91 15116 00 Orange Filter 590 50 nm EBR 500K Red Filter 620 40 nm SYPRO 500 Green Filter 537 35 nm 91 13840 00 Blue Filter 460 40 nm HB 500 AlphaView Stand Alone software 91 14425 00 Imaging Test Target 85 13532 00 Magnetic Stylus 85 14195 00 Digital Thermal Grayscale USB Printer 91 12999 00 Thermal Paper High Gloss High Contrast 5 rolls P 200H Thermal Paper High Contrast 4 rolls P 150H Spares Service Parts 302 nm UV Replacement Bulb 75 14084 00 365 nm UV Replacement Bulb 75 14083 00 Fuse 250 V 6 A 011 597 FluorChem E System User Guide page 120 Chapter 10 Maintenance and Troubleshooting Part Description Part Number Service Contracts FluorChem E Platinum Service Contract S FCE PL Fluorchem E Gold Service Contract S FCE GS FluorChem E Silver Service Contract S FCE SS Fluorchem E Upgrade to Platinum Service FluorCche
52. er exposure to UV light As a safety precaution the UV light will automatically turn off after three minutes when using the inter lock switch General Guidelines and Information System Operation Notification of Intended Use NOTE The FluorChem E system is for research use only Not for use in diagnostic or clinical procedures FluorChem E System User Guide System Specifications page 21 Lifting and Moving the System Use Proper Lifting Precautions IMPORTANT Use caution when lifting or moving the Fluorchem E system The system weight is 75 Ibs 34 ko Two people are required to lift the system onto a laboratory bench Proper lifting safety precautions must be taken System Specifications Description Specification Dimensions 62cm Dx62 cM Hx 32 cm W 24 5 in x 24 5 in x 12 6 in Weight 34 kg 75 Ibs Power Requirements 100 230 V AC 50 60 Hz self adjust ing Power Consumption 345 W Operating Humidity Range 15 70 relative non condensing Operating Temperature Range 15 25 09 77 F Detector 8 Megapixel scientific grade CCD 16 bit A D thermoelectrically cooled to 25 C absolute and regulated Field of View 16 cm x 12 cm fixed For indoor use only FluorChem E System User Guide page 22 Chapter 3 System Hardware FluorChem E System User Guide page 23 Chapter 4 Getting Started Chapter Overview Powering on the System Overview of FluorChem E User Interface Imaging Setup UV Light Overri
53. f skin that may be exposed to UV light when engaging the UV interlock switch override Filter Wheel The FluorChem E system is equipped with a motorized 4 position filter wheel The system comes with a pre installed 590 50 nm orange filter Commonly used for ethidium bromide or SYPRO orange applications In addition the following optional filters are also available 620 40 nm red filter for SYPRO Red SYPRO Ruby or Deep Purple 537 35 nm green filter for SYBR Green SYBR Gold or SYBR Safe 460 40 nm blue filter for Hoeschst Blue DAPI ANS or Methylcoumarin For information on how to insert optional filters into the filter wheel please refer to Installing Emission Fil ters on page 116 If new filters are installed or existing filters are moved to different positions on the filter wheel the filter settings in the System Settings Screen must be updated accordingly To view or change filter settings please refer to Changing Filter Settings on page 91 Computer The FluorChem E system contains an integrated computer with a 250 GB hard drive Acquired images are automatically archived on the hard drive Access to network and side USB ports is located on the right side panel near the rear of the instrument Network Port The 10 100 BASE T Ethernet RJ 45 connector can be used to connect the FluorCchem E system computer to a local network Please see Chapter 9 Networking and Remote Access for information on accessing
54. filter select Filter Settings box and select the desired option FluorChem E System User Guide Overview of the Darkroom Screen page 41 Channel 1 NolLight No Filter 1 Orange 590nm Q Green Sarnm a Q Red 620nm i NOTE Up to three filters can be installed in the FluorChem E system The Filter Settings box will only display installed filters For information on how to add or change the filters listed in the Filter Settings box please refer to Changing Filter Settings on page 91 Exposure Settings To change exposure settings select the Exposure Settings box and select one of the following options Channel 1 7 Chamet No Light No Filter Auto D Manual Auto Exposure When Auto is selected in the protocol the FluorChem E system will take successively longer exposures dur ing image acquisition until the optimal exposure is reached However users can select the Stop button at any time to save and display the last completed image exposure taken FluorChem E System User Guide page 42 Chapter 5 Image Acquisition Manual Exposure When Manual is selected in the protocol the Fluorchem E system will take one image at the exposure time entered To enter an exposure time select the time box A numeric keypad will appear to allow the user to enter a time setting Any value between 50 ms and 3 hours can be used Settings are displayed in either seconds or minutes To e
55. for exam ple 2010 06 16_09 22 15_1_16bit png However ProteinSimple recommends changing the default title to a unique entry as this will help identify archived image files Printing Images Images saved on the Fluorchem E system can be printed to a digital grayscale thermal USB printer Please contact ProteinSimple Technical Support to obtain information on compatible printers To print an image select the Print button The system will first confirm a printer is available and then display the following dialog box Print to attached printer x Select the Print button from the dialog box Printouts will contain the title and project entered by the user in the Image Info page as well as the system and protocol information stored with the image Protocol settings Date and time of image acquisition File name Instrument name and location The following is an example of an image printout FluorChem E System User Guide page 70 Chapter 6 Image Review Channel Date and Settings gt No Light 1 00m No Light 30s No Light 30s 2011 03 01 01 33PM 4 Time Title 2011 03 01_ 13 33 06 Protein Study lt Project Notes gt Initial data set FluorChem E FE0232 Laboratory FCE 0232 _2011 03 01_13 33 06 fcz Instrument Name File Name and Location FluorChem E System User Guide Copying Printing and Deleting Images page 71 Deleting Images To delete an image select the Delete button The following dialog
56. g box FluorChem E System User Guide Chapter 5 Image Acquisition Chapter Overview Overview of the Darkroom Screen Image Acquisition Sample Imaging FluorChem E System User Guide page 35 page 36 Chapter 5 Image Acquisition Overview of the Darkroom Screen Selecting the Darkroom Tab will display the Darkroom Screen his screen is used to view protocols and channel settings and for image acquisition Imaging Setup The Setup page will be displayed whenever the Darkroom Tab is selected Users can select protocols change channel settings and start image acquisition in the Setup page Show Hide Channels Button Darkroom Tab TOCE Image Display _ 4 Protocol Menu b Chemiluminescence Expose Button eo Expose Image Display The Image Display area will display a white light image that is automatically taken to help with sample posi tioning prior to image acquisition Protocol Menu This menu allows users to select from a set of pre programmed protocols Protocol options can be viewed by selecting the Protocol Menu box FluorChem E System User Guide Overview of the Darkroom Screen page 37 Chemiluminescence X Chemiluminescence Chemi with Markers Colony Plates Coomassie Blue Ethidium Bromide Silver Stain SYBR Green Safe Show Hide Channels Selecting the Channel Show Hide button to the left of the Protocol Menu box will display the channels use
57. h the amount of room required FluorChem E System User Guide Backup and Restore page 85 During the backup the status appears Backup and Restore x Copying to USB 3 of 10 Images o Cancel When backup is completed the following dialog box appears Backup and Restore x Backup Completed BACKUP_FEO232_2012 06 06 It is now safe to remove the USB drive Close Restore Insert the USB containing backup files The instrument will search the USB drive for the backup folders but will only show the latest backup To restore from an earlier backup the more recent backups need to be removed from the USB The Start Restore dialog box reports how many backed up images the folder contains The term images refers to runs which may have multiple images each All existing runs are maintained during the restore If a backed up run already exists on the instrument it is skipped in the restore so changes to a run since the backup will be maintained If a run restored to the album page is also in the trash the copy in the trash is deleted FluorChem E System User Guide page 86 Chapter 7 Managing Stored Images During restore the status appears Backup and Restore x Restore from USB 1 of 9 Images Cancel When restore is completed the following dialog box appears Backup and Restore x Restore Completed It is now safe to remove the USB drive Close FluorChem E S
58. he Save button FluorChem E System User Guide page 90 Chapter 8 System Settings Name and Location P Nal Part hamn E FEO232 SN FE0232 Location Laboratory Changing Date and Time Settings To change the date time or time zone setting select the Time button The following settings will display Date Time Box Boxes Set Time Tuesday 25 January 2011 12 31 PM aii jin r GMT 8 00 Pacific Time US amp Canada t Time Zone Selection To change the date select the Date box and choose a new date from the calendar provided The current date will be displayed in yellow Use the arrows at the upper left and right to change the calendar month January 2011 o Su Mo Tu We Th Fr Sa Dhoom Bled Sih oz el 9 10 11 12 43 44 15 16l 17 18l 19 20 21 22 23 26 27 28 291 30 31 To change the time select hours or minutes in the Time boxes and use the number pad to enter the new information FluorChem E System User Guide Changing Filter Settings page 91 9 o8 Am North America del OK TH LEJ F9 Ay a KG L ee HS 0 The AM or PM setting can be changed by selecting the AM PM menu box The current setting will be dis played in orange text PM AM PM To change the time zone select the Time Zone menu box and choose an option from the drop down list The current setting will be displayed in orange text GMT 8 00 Pacific Time US a
59. hem E System User Guide page 128 Chapter 11 Analysis in Digital Darkroom Band Sensitivity The sensitivity of the software in detecting bands can also be set using the Slider bar FluorChem E System User Guide Summary Table page 129 Summary Table The summary table includes the Lane number band Position signal Intensity and Total each band contributes to the intensity of a lane For multi color images reads from each wavelength will be color coded e cLane By default the lanes are numbered starting with 1 on the left The lane title can be edited in the table and will be displayed above the lane in the image e Position The position if each band is reported as a percentage of the height of the selected region Selecting a ladder lane will change the Position column to a molecular weight MW column Intensity Represents the total band intensity Total Contribution of each band to the total channel intensity for the lane The sum of all the bands in each channel is 100 within the rounding error Lane Position Intensity 5T otal F 33 3 166738 100 0 a 32 9 73883 100 0 3 30 0 62470 5 7 33 8 547217 50 2 37 4 366166 33 6 39 8 43586 4 0 48 9 32317 3 0 52 5 37722 3 5 ra 20 7 39135 7 3 35 0 474025 89 0 39 3 19454 37 5 32 1 21663 100 0 6 3 2 83183 6 9 15 8 82285 6 8 25 4 75950 6 3 34 3 176731 14 7 52 8 236280 19 7 67 6 148718 12 4 80 4 191877 16 0 94 2 206699 17 2 FluorChem E System User Guide
60. iew image data for a single channel toggle the Image Show Hide button next to the desired Channel Settings box to Show Next toggle the Image Show Hide button s for the remaining channels to Hide In the example that follows image data for only Channel 1 is shown Channel 2 image data is hidden FluorChem E System User Guide Channel Image Display and Overlay page 61 Chemi with Markers Info Adjust gt Channell A gt Channel 2 Analysis Overlaying Channel Image Data To overlay image data for multiple channels toggle the Image Show Hide buttons next to the desired Channel Settings boxes to Show If needed toggle the Image Show Hide button s to Hide for any unwanted channels When multiple channels are toggled to Show the channel image data will be overlaid automatically In the next example image data for Channel 1 and Channel 2 are shown overlaid NOTE When the Chemi with Markers protocol is used for image acquisition the chemiluminescent image of the sample Channel 1 will display in grayscale and the white light image of the markers Channel 2 is displayed in red to optimize visualization FluorChem E System User Guide page 62 Chapter 6 Image Review Chemi with Markers gt Info Adjust _ a v Channell A Channel 2 6 Analysis Making Image Adjustments NOTES Image adjustments are automatically saved to the image file as the enhancement settings ar
61. il in the next chapter FluorChem E System User Guide page 55 Chapter 6 Image Review Chapter Overview Overview of the Review Screen Entering Image Information Channel Image Display and Overlay Making Image Adjustments Image Zoom Viewing Pixel Values Browsing Images FluorChem E System File Information Copying Printing and Deleting Images FluorChem E System User Guide page 56 Chapter 6 Image Review Overview of the Review Screen Selecting the Review Tab will display the Review Screen This screen is used to view images protocol and channel settings enter or edit image information and make any needed enhancements Stored images can also be deleted copied and printed The Review Screen will be displayed automatically when image acquisition has completed NOTES Images are displayed overlaid by default for protocols that acquired images on multiple channels Forimages acquired using the Chemi with Markers protocol the white light image of the markers Channel 2 is displayed in red to optimize visualization Images for channels acquired using all other protocols and or light sources will display in grayscale Date and Time of Review Tab Image Acquisition Darkroom MB Review E Images File p 2012 05 01_09 32 57 May 01 2012 9 32 AM same Protocol fo Edit e _ Image 201 2 05 01_09 32 57 Image Information Review Display Adjust Adjust Cha
62. ilter Wheel 09 36 03 PASSED Apr 06 2011 11 11 ABORTED Aperture 09 36 20 PASSED Focus 09 37 07 PASSED Camera 09 37 07 PASSED UV 09 37 20 PASSED White LED 09 37 40 PASSED Dark Masters 09 37 40 PASSED Flat Field Masters 09 37 40 PASSED A Test Log of previously run self tests with overall pass fail results is shown on the left To view results for a particular test select the desired Test Date in the log The Results Table will display the start time individual test result and failure reason for each component tested FluorChem E System User Guide Creating Dark Masters page 97 Running the Self Tests 1 Open the FluorChem E system door and remove any sample screens from the UV transilluminator 2 Close the system door 3 Select the Run Test button FluorChem E will automatically perform individual component tests and report results Testing takes approximately 5 minutes to complete To copy test file results insert a USB drive into any USB port on the instrument and select the Copy button NOTE If any of the tests return a failed result please contact ProteinSimple Technical Support Creating Dark Masters NOTES Dark masters are generated on the FluorChem E prior to system shipment Dark Master files typically only need to be updated as required for system service Dark Master file generation requires approximately 12 hours of uninterrupted system time during which the FluorChem E may not be used for image acquisition Prot
63. ing exposures will display Exposure 1 Raw image with no exposure corrections Exposure 2 Image at optimum contrast with automatic leveling of black and white values so the image is displayed using full gray levels of the screen or monitor This exposure is selected by default Exposures 3 9 Display views of increasing exposure levels from Exposure 2 FluorChem E System User Guide page 64 Chapter 6 Image Review Darkroom B Review E images Chemiluminescence Info Exposures 1 3 gt Adjust Exposures 4 6 gt Channeli o ad Analysis Exposures 7 9 gt Select the image with the desired exposure The current exposure selection will be highlighted in dark blue The exposure selected in this view is used as the default image display for the Review and Images screens and will be the default 8 bit ong image stored with the fcz file To choose a different exposure as the default image simply select the desired exposure To exit the page select the Exposures View button again Image Zoom The Review Screen also allows users to enlarge the size of the displayed image using the Zoom Q option The Zoom button can be toggled between 200 400 and Off Off 200 400 Zoom Zoom To enlarge an image toggle the Zoom button to the desired magnification level To view other areas of the enlarged image select and drag the image until the desired viewing area is shown To exit Zooming
64. irements The FluorChem E system label displays the system power specifications se K 0 proteinsimple Santa Clara CA USA Model FluorChem E Made in USA OOVAC 230VAC 50 60Hz 345 Watts _ rust T6 3AH 250V n lt lt In addition to the electrical power requirements listed above ProteinSimple requires that all grounded circuits terminate at the receptacles and the receptacles must be located within 10 ft 3 m of the instrument FluorChem E System User Guide page 10 Chapter 2 Installation Environmental Requirements Please provide appropriate heating and cooling to maintain a constant laboratory temperature For optimal FluorChem E system performance the laboratory environment must meet the following criteria Requirement Specification Operating temperature range Pea G77 Operating humidity range 15 70 relative non condensing Table 2 3 Fluorchem E system environmental requirements Software and Computer Requirements The FluorChem E system includes an integrated computer with the FluorChem E User Interface pre installed Post acquisition image analysis can be performed using AlphaView stand alone software Please use the AlphaView software CD provided with the FluorChem E system to install the software on a separate computer workstation such as a user s desktop computer In order to run AlohaView software this computer must meet the minimum requirements listed in the table below
65. key to delete an existing entry When text entry is complete touch the Save button FluorChem E System User Guide Overview of FluorChem E User Interface page 27 User Interface Screens The FluorChem E User Interface has four main screens that are accessed through the following tabs Dark room Review Images and System Settings Each screen can be accessed by selecting the tab on the touch screen System Settings Tab oo Darkroom Screen Selecting the Darkroom Tab will display the Darkroom Screen This screen is used for protocol setup and image acquisition Chemiluminescence 8J Expose FluorChem E System User Guide page 28 Chapter 4 Getting Started Review Screen Selecting the Review Tab will display the Review Screen This screen is used to view stored images and protocol information enter or edit image details and make image enhancements Images can also be deleted copied and printed a Darkroom E Review 2012 05 01_09 32 57 May 01 2012 9 32 AM Ethidium Bromide Info Edit 201 2 05 01_09 32 57 Adjust fj Channel 1 amp 441 of 441 4 Older p Q FluorChem E System User Guide Overview of FluorChem E User Interface Images Screen page 29 Selecting the Images Tab will display the Images Screen This screen is used to browse saved images Stored images can also be deleted and copied from this screen uam Album
66. lete the software update FluorChem E System User Guide Saving Custom Protocols page 95 Saving Custom Protocols Custom protocols can be created and stored for future use These can be accessed through the drop down protocol menu see page 38 1 To create a protocol select Protocols The User Protocols screen allows for both the creation of new protocols and modification of existing protocols Factory supplied protocols cannot be edited User Protocols Save Protocals 2 Select New to bring up the protocol details The protocol name and read settings can be entered here FluorChem E System User Guide page 96 Chapter 8 System Settings User Protocols Save Protocols Details Protocol 1 Protocol 1 Channel o At Q Epi white p No Filter wy Manual 1 seconds YS Standard Channel 2 g Channel 3 oy Delete New EE z 3 Click Save to save the User Protocol or Cancel to discard changes 4 Protocols can be deleted by selecting the protocol and then Delete System Self Test FluorChem E system performance can be tested to ensure key components are functioning properly To run the system self tests or view results from previous tests select the Test button Self Tests Mt BEL TSE a Ep 6 te 2 4 rw ee ee Results Test Log Test Date Result Name Start Result Failure Reason lt a Table Apr 08 2011 12 11 PASSED System 09 35 42 PASSED Apr 07 2011 09 35 PASSED F
67. m E Upgrade to Gold Service Preventive Maintenance Field Service Preventive Maintenance Depot Service Troubleshooting Please contact ProteinSimple Technical Support at 888 607 9692 or support proteinsimple com for trou bleshooting service or support related items FluorChem E System User Guide page 121 Chapter 11 Analysis in Digital Darkroom Chapter Overview Lane Profiling Summary Table Renaming Lanes Adding Molecular Weight Ladders Assigning Molecular Weight Ladders Exporting Printing FluorChem E System User Guide page 122 Chapter 11 Analysis in Digital Darkroom Lane Profiling Lane profiling analysis is available through the Digital Darkroom software running on the FluorChem E It can be used to measure densitometry or to determine molecular weights of samples Locating Analysis Tool When an image is selected for Review the Analysis tool becomes available Click on the Analysis button to begin Review Darkroom cet eerie Aj 1 2007 9 01 AM Coomassie Elie Into Edit fi fc i 44 Adjust a Channel g e Analysis lt _ Analysis 15 of 72 kler Newer Ol FluorChem E System User Guide Lane Profiling page 123 Defining Number of Lanes Clicking on Analysis expands the window The next step will be to define your lanes To do this click on Define Lanes Analysis Define Lanes The number of sample lanes can be defined by entering th
68. mide Channel 1 Channel 1 QO No Light i HUI Trans UV No Filter z Q Orange 590nm 1 Auto O Auto i aa Standard Ez Standard E Full Resolution Full Resolution Standard Resolution When Standard Resolution is selected in the protocol the FluorChem E system will acquire the image with a 4 x 4 pixel bin 832 x 626 pixels for chemiluminescence protocols and a 3 x 3 pixel bin 1109 x 835 pixels for all other protocols Standard Resolution is the default and recommended setting forallimaging protocols Forchemiluminescenceimaging this setting significantly decreases the exposuretime required to visualize samples with low signal intensities Full Resolution When Full Resolution is selected in the protocol the FluorCchem E system will acquire the image using full 8 megapixel resolution 3326 x 2504 pixels Full Resolution is only recommended when publication qual ity and large printouts e g poster size are required or to resolve extremely small feature sizes lt 150 microns Selecting Full Resolution significantly increases the exposure time required compared to Stan dard Resolution NOTE Selecting Full Resolution will result in increased exposure time This increased exposure time can be very significant particularly when imaging low abundant chemiluminescent targets FluorChem E System User Guide Overview of the Darkroom Screen page 45 Multi Channel Imaging FluorChem E provides optional multi ch
69. milar to using the touchscreen interface directly on the FluorChem E system except for the following functions Text entry The keyboard on the remote computer or device is used for all text and numeric entry Printing The Print command will print image files to a local or network printer only Copying Image Files In the remote interface an option to download files is provided instead of the copy option Downloading Image Files Stored image files can be downloaded to the remote computer or device Users are provided the option to either download the fcz file or just the ong files NOTE Depending on the option selected downloaded files can be provided in zip format WinZip or a similar extraction program is required to extract zip files To download files 1 Select the Download button in the Review Screen or from an Edit page in the Images Screen NOTE Multiple images can be selected for download in the Album Edit or Trash Edit pages in the Images Screen 1 The following dialog box will display FluorChem E System User Guide Downloading Image Files page 107 i AlphaView Open in AlphaView fcz files Images Images png files Results Results table csv file Cancel 2 Choose a download option Select the AlphaView button to download the cz file If desired this option can launch AlphaView software automatically after download Select the Images button to download only the 8 and 16 bit ong image file
70. mote ACCESS nn 104 CONNCCUNG TOHIUOICNOM ode cece neemustae cows 104 POW PIOCGINOWM GAGS FCS i a tins sete steel eat t ast 106 Opening Image Files in AlphaView Software 108 ODN FOZ MOS EE E bes ate doer tial 108 Ovena ONG FICS lt 6 52st ttt S 110 Chapter 10 Maintenance and Troubleshooting 111 MENON O er sen Ge a RE 1712 Cleaning the Fluorchem E System 112 UV Transilluminator Bulb Replacement Bre MSti ERIS HOM FIE see e E 116 CHEN GING ET U E riere a En 118 Accessonesand Spare PONS 2 saciccsdasics eae us 119 VOGO OO TIVO ede ere an hes ese a as tect eats 120 Chapter 11 Analysis in Digital Darkroom 121 PGC VOM e EEEE E E eth cdma inl EEE 122 LOCOUMOAIIGIVSIS TOO an irn Btls 122 Denning Numeral Lanes areires reer 123 DETINING REGION OF INTEPESE eriein conned 124 Rotating Region of Interest 0005 125 Calcuding eny Val S san a 126 DINE C O Ba erste E tera Rote etd 126 Adding and Removing Individual Bands 127 PAG EI N a aa r a 128 E ANIE A E EE EEEE ES 129 Renaming TONGS eaaa EE AA ERA 130 Adding Molecular Weight Ladders 4 130 FluorChem E System User Guide page iii Assigning Molecular Weight Ladders 131 PPO aaret aa aA eae at A E 133 FAND enire rearea chhieinta 134 FluorChem E System User Guide page 1 Chapter 1 General Information Chapter Overview Welcome Introduction to the FluorChem E System Safety Customer Service an
71. mp Canada Eastern Standard Daylight Time North America Western European Time Summer Time Central European Time Summer Time Eastern European Time Summer Time GMT 12 00 Eniwetok Kwajalein GMT 11 00 Midway Island Samoa GMT 10 00 Hawaii GMT 9 00 Alaska GMT 8 00 Pacific Time US amp Canada Da When the desired date and time settings have been updated select the Save button to save the changes Changing Filter Settings To change filter settings select the Filter button This will display the type of filters currently used in the Fluorchem E system and their locations in the filter wheel FluorChem E System User Guide page 92 Chapter 8 System Settings Filters Installed Save en e No Filter Q Orange Red Position Boxes Q Green Position 1 does not contain a filter and is used whenever a protocol uses the No Filter option The Position 1 setting cannot be changed by the user Positions 2 4 can be set by the user and should reflect what filters are currently installed in the filter wheel To change the filter setting for a particular filter position select a Position box and choose a filter from the drop down list Q No Filter 2 Empty Red Q Orange h 4 Q Green j Blue ae ee If the position does not contain a filter the setting should be set to Empty Filter options for each position include orange red blue green or empty When filter settings have b
72. necessary if the system power remains on at all times If the system optics temperature is not at optimum 25 C when acquisition is started a message will be displayed providing users with the option to either cancel or proceed with the acquisition To image samples using the FluorChem E system 1 Place the sample on the appropriate sample screen and center it on the imaging area 2 Open the system door The Door Open Screen will display FluorChem E System User Guide Sample Imaging page 53 Colony Plates Chemiluminesence Colorimetric Center blot on black screen Center gel on clear screen Center sample on white screen Place screen on light box then close door f FluorChem E FE0232 Laboratory http fe0232 local ph egg oe ee 3 Place the screen on the UV transilluminator 4 Close the system door The FluorCchem E system will take an epi white light image of the sample auto matically 5 Ensure that the area of interest in the blot or gel has been captured in the image If not open the system door and reposition the sample in the imaging area of the sample screen as needed The FluorChem E system will take a new white light image each time the system door is closed 6 Select a protocol from the Protocol Menu change settings in the Channel Settings Menu make changes to image information in the Info box and add channels if desired Select the Expose button to start image acquisition The A
73. nnel 1 Channel Information 441 of 44 lt ____ mage 4 Older b Q Number Ih o Image Review Options FluorChem E System User Guide Overview of the Review Screen page 57 Image Review Display The Image Review Display will display saved sample images The file name is located above the display on the left and the date and time of acquisition is located above the display on the right The image number is shown below the display on the right Image Enhancement Tools The enhancement tools on the Review page under Adjust include Brightness Controls Exposures View and Invert The use of these tools will be discussed in more detail later in this chapter lt Exposures View Brightness Controls 0 F 4 Invert Protocol Information A copy of the acquisition protocol is stored with all images The Protocol box will display the name of the acquisition protocol used for the image currently displayed Channel Information The Channel Settings box es will display the individual channel settings used for image acquisition To view light filter exposure and resolution settings select the Channel Settings box Channel 1 lt UUI Trans UV Time 920ms fim Standard FluorChem E System User Guide page 58 Chapter 6 Image Review NOTE If the Auto Expose setting was used and image acquisition was stopped by the user Auto Stopped will display as the exposur
74. nter a setting of less than 1 second select the decimal point first Example 1 Manual Exposure Time of 50 ms Enter 05 for a manual exposure time of 50 ms The time displayed will be 0 05 seconds Chemiluminescence Channel 1 QO No Light fl Be Got dei No Filter 4 c fe J Manual 1 2 3 10 seconds Fel minutes O OK Sa Stanuara FluorChem E System User Guide Overview of the Darkroom Screen page 43 Example 2 Manual Exposure Time of 90 seconds Either 130 or 90 00 can be entered for a manual exposure time of 90 seconds The time displayed will either be 1 30 minutes or 90 seconds depending on how the time was entered Chemiluminescence 4 Chemiluminescence Channel 1 Channel 1 NoLight No Light 8 9 gael No Filter t No Filter Aa Bel 6 A Manual f YE Manual 1 2 3 Time 1 30 minutes Time 90 00 seconds 0 OK 5u Standard 5u Standard Example 3 Manual Exposure Time of 3 hours Enter 18000 for a manual exposure time of 3 hours The time displayed will be 180 minutes x Chemiluminescence A Channel 1 NoLight 8 9 del No Filter A e 6 ws Manual 1 2 3 Time 180 00 minutes 0 S 5u Standard FluorChem E System User Guide page 44 Chapter 5 Image Acquisition Resolution Settings To change resolution select the Resolution Settings box and select one of the following options Chemiluminescence x Ethidium Bro
75. ol name Date and time of image acquisition e Protocol exposure and resolution settings FluorChem E System User Guide page 76 NOTES Chapter 7 Managing Stored Images Multiple will display for exposure settings when image files contain channel data with different exposure times NOTE If the Auto Expose setting was used and image acquisition was stopped by the user Auto Stopped see protocol details will display as the image title 2011 03 01_15 16 56 Ethidium Bromide 03 01 2011 270ms Standard 3 16PM 2011 03 01_13 48 41 Ethidium Bromide 03 01 2011 630ms Standard 2 35PM 2011 03 01_13 38 13 Chemiluminescence viet be 2011 40s Standard 2011 03 01_13 33 06 Protein Study Chemiluminescence pe be 2011 Multiple Standard 201 1 03 01_13 30 54 Chemiluminescence red 2011 Multiple Standard 1 30PM 2011 03 01_12 08 48 Protein Study ERK2 Chemiluminescence eas 1 Multiple Standard PC 2011 03 01_12 02 50 Protein Study ERK1 Chemiluminescence a oe 1 10s Standard 2011 02 24_12 40 57 Ethidium Bromide 02 24 2011 920ms Standard 12 40PM 2011 02 24_10 20 08 Chemi with Markers 02 24 2011 Multiple Standard 10 20AM 2011 02 23_11 43 41 Coomassie Blue 02 23 2011 670ms Standard 11 43AM 20110222 100525 Coomassie Blue 02 22 2011 Multiple Standard 10 05AM 2011 02 21_16 29 18 Coomassie Blue 02 21 2011 560ms Standard 4 29PM By default image files in the Table view are sorted by date with the most recent files at
76. operly on the UV transilluminator Sample Screens The FluorChem E system is provided with the following screens for sample imaging The screen used depends on the application Black screen Chemiluminescent and epi white light imaging Clear screen U V excited gel imaging White screen Colorimetric gel imaging and colony plates Blue screen optional Blue excited fluorescent gel imaging Sample Positioning To position samples in the FluorChem E system 1 Place the sample on the appropriate sample screen and center it on the imaging area 2 Open the system door The Door Open Screen will display Darkroom Colony Plates Colorimetric Chemiluminesence Center blot on black screen Center gel on clear screen Center sample on white screen Place screen on light box then close door i FluorChem E FE0232 Laboratory http fe0232 local Pa Sa FluorChem E System User Guide page 32 Chapter 4 Getting Started J Place the screen on the UV transilluminator 4 Close the system door The FluorChem E system will automatically take an epi white light image of the sample Chemiluminescence proteinsimple e Expose Ensure that the area of interest in the blot or gel has been captured in the image If not open the system door and reposition the sample in the imaging area of the sample screen as needed The FluorChem E system will take a new white light image each time the syst
77. ottle Spray the cleaner onto the cloth and gently wipe the screen in a circular motion Do not press hard while cleaning the screen Continue to wipe until all of the solution is removed and the screen is completely dry NOTE Leaving excess cleaning solution on the LCD touchscreen may cause long term damage to the screen System Decontamination The following decontamination procedure must be performed should the FluorChem E system come in con tact with hazardous samples Items needed 10 bleach solution and a soft cloth or paper towels 1 Wear rubber gloves 2 Completely saturate a paper towel or cloth with 10 bleach solution FluorChem E System User Guide Maintenance page 113 3 Wipe down the surface of the UV transilluminator sample screens and any other areas inside or outside the system that may have come into contact with hazardous samples 4 Wipe down the exterior of the instrument UV Transilluminator Bulb Replacement NOTE To ensure uniform illumination ProteinSimple recommends replacing all six UV transilluminator bulbs at the same time 1 Power down the FluorChem E system and unplug the power cord from the system 2 Open the system door and slide out the UV transilluminator approximately halfway 3 Using a Philips head screwdriver unscrew the four screws on the UV transilluminator outer front panel and remove the panel This will expose the inner front panel FluorChem E System User Guide page 1
78. ow the instructions in the browser dialog box to download the fcz file When download is complete AlohaView software will automatically launch and open the file NOTE Some browsers may prompt for the location of the AlphaView software exe file on the remote com puter If this occurs select the Browse option in the browser dialog box Browse to where AlphaView soft ware is installed such as Program Files ProteinSimple AlphaVIEW SA and select the AlC AlphaView exe file e Opening saved or extracted files on the remote computer This can be done in one of two ways Browse to the folder where the file is located and double click the desired fcz file to automatically launch AlphaView software and open the file Open AlphaView software Select File gt Open Browse to the file location and select the desired fez file In AlohaView software the displayed fcz file can be renamed and saved as a 16 bit tif file To do this select Save As from the File menu FluorChem E System User Guide Opening Image Files in AlphaView Software page 109 AlphaView Alpha VIEW SA The following dialog box will display In the Save as type drop down menu select Tiff Files tif The original file name can be changed if desired x Se ee Search Dest p r ATE Name i Item type Date modified EE Desktop Libraries J Downloads T Recent Places _ FCE_2010 06 16_09 File folder 6 24 2010 11 28 Ab Libraries coomassie tif 80KB
79. page 1 So proteinsimple Formerly Cell Biosciences Fluorchnem E System User Guide Copyright 2012 ProteinSimple All rights reserved ProteinSimple 3040 Oakmead Village Drive Santa Clara CA 95051 Toll free 888 607 9692 Tel 408 510 5500 Fax 408 510 5599 email support proteinsimple com web www proteinsimple com FluorChem E System User Guide FluorCchem E User Guide P N 94 15038 00 Revision 4a June 2012 For research use only Not for use in diagnostic or clinical procedures Patents and Trademarks Automatic Image Capture AIC and digital ProteomeChip technology are covered by U S Patent Nos 6 995 901 6 909 459 6 853 454 6 271 042 7 166 202 and other issued and pending patents in the U S and other countries ProteinSimple the ProteinSimple logo the Aloha Innotech logo Protein Forest the Protein Forest logo the AIC logo AlohaCal Alohalmager AlphaPart1 1View AlohaQuant AlohaSnap AlohaSpec AlphaUV AlphaView ChemiGlow Chromalight dPC digital ProteomeChip Fluorchem MSRAT Multilmage NanoPro the intertwined helix design iWB ProteomeChip red SpectraPlex Xpedition XplorBright and XplorUV are trademarks or registered trademarks of ProteinSimple Other marks appearing in these materi als are marks of their respective owners FluorChem E System User Guide Table of Contents Chapter 1 General Information ceeeees 1 WV NCO il Seth eit re Rieti Cnet ah ieee pane
80. roducts For questions or general feedback use the following contact methods FluorChem E System User Guide page 6 Telephone 408 510 5500 888 607 9692 toll free Fax 408 510 5599 E mail Support proteinsimple com Web www proteinsimple com Address ProteinSimple 3040 Oakmead Village Drive Santa Clara CA 95051 USA Legal Notices NOTE Read the Legal Notices carefully before using your FluorChem E system FluorChem E System Disclaimer of Warranty Chapter 1 General Information EXCEPT AS EXPRESSLY PROVIDED IN ANY ProteinSimple SOFTWARE LICENSE AGREEMENT OR QUOTATION THE PRODUCTS SOLD AND SERVICES PROVIDED BY ProteinSimple ARE PROVIDED ON AN AS IS AND AS AVAILABLE BASIS WITHOUT WAR RANTY OF ANY KIND ProteinSimple AND ITS SUPPLIERS DO NOT WARRANT THE SECURITY PRIVACY OR ACCURACY OF ANY DATA PROVIDED VIA THE PRODUCTS OR SERVICES AND YOU AGREE THAT THE USE OF ANY SUCH DATA BY YOU IS AT YOUR SOLE RISK TO THE MAXIMUM EXTENT ALLOWED UNDER APPLICABLE LAW ProteinSimple AND ITS SUPPLIERS DISCLAIM ANY AND ALL WARRANTIES WHETHER EXPRESS IMPLIED OR STATUTORY INCLUDING WITHOUT LIMITATION ANY WARRANTY OF MERCHANTABILITY TITLE NON INFRINGEMENT OR FITNESS FOR A PARTICULAR PURPOSE FluorChem E System User Guide page 7 Chapter 2 Installation Chapter Overview Introduction Space Requirements Electrical Requirements Environmental Requirements Software and Computer Requirements System
81. rt and expected completion times will be shown along with an image counter and progress bar which will update during the calibration process 3 When the UV white light flat field calibration has finished or was skipped the following message will dis play to indicate the process is complete Flat Field Masters created m Ea gt Create Flat Field Masters Click OK to exit Changing Focus Settings NOTE The FluorChem E system optics are set to an optimal fixed focus above the sample screen and do not normally need to be changed However if the focus settings do need to be adjusted use the following procedure 1 Place the imaging test target provided with the FluorChem E system on the black sample screen and cen ter the target on the imaging area 2 Place the screen on the UV transilluminator FluorChem E System User Guide Changing Focus Settings page 101 J Close the system door A white light image of the target will automatically be taken and displayed 4 Go to the System Settings Tab and select the Focus button The Focus Screen will appear and the sys tem will automatically take and display new white light image NOTE White light focus images may be blurry until optimum focus has been reached Adjust the focus incrementally by selecting the Focus Adjustment buttons until the optimum focus is reached The two middle buttons single arrows will adjust the focus up or down in minor increments
82. rument icon in blue text The Fluorchem E system name and location are included on image printouts and are also used to identify the instrument on the network System name and location information can be customized by the user by clicking on the Name button The default system name will include the serial number for example FEO232 FluorChem E System User Guide Changing System Information page 89 Software and Network Information The current versions of the FluorChem E Digital Darkroom software and firmware are displayed in the upper right corner of the screen 2 0 118636 1 6 Network information is located just below the version information and displays the FluorChem E system net work status IP and MAC addresses Network with DHCP 10 1 1 185 00 18 7d 0f 25 b8 NOTE Ifthe FluorChem E system is not connected to a local network the Network Status will display local connection no network Optics Temperature The current temperature of the FluorChem E system optics is also shown in the System Settings Screen The temperature displayed is a static measurement taken at the time the screen is accessed To get an updated reading at any time select the Refresh button Refresh Or 25 0 C CG Button Changing System Information To change the FluorChem E system name or location setting select the Name button Select either text box and use the keyboard to enter the new information When text entry is complete select t
83. rview of FluorChem E User Interface This section will provide a general overview of the FluorChem E User Interface and touchscreen operation Using the Touchscreen The FluorChem E User Interface is accessed through the touchscreen Users can initiate acquisition access protocols and images and change system settings via touchscreen commands Tabs Tabs are used to access particular screens in the Fluorchem E User Interface To view a screen touch the screen tab Buttons Buttons are used for direct instrument control To initiate a command touch the button eo Expose Menu Boxes Menu boxes contain a list options that can be selected by the user Chemiluminescence To view the items in the menu box drop down list touch the box To select an item in the menu simply touch the item FluorChem E System User Guide page 26 Chapter 4 Getting Started Chemiluminescence Chemiluminescence Chemi with Markers Colony Plates Coomassie Blue Ethidium Bromide Silver Stain SYBR Green Safe Text Boxes Text boxes display information typically entered by the user To type an entry into a text box touch the box A keyboard or number pad will display on the screen and a cursor will be placed in the text box Use the keyboard or number pad to type in the entry Save Cancel Bil IAA E FR e ere lel abia D pF Al Se Del be el 4G eA bd Ket shift Zol Kel eG e MeL Be NEM del 123 return Use the del
84. s NOTES If more than one image is selected when using the AlphaView download option a single zip file will be downloaded that contains all the fcz files for the selected images Ifonly one image is selected a single fcz file will download When using the Images option all png files for an image are provided in a single zip file 3 Follow the instructions in the browser dialog box to download the file s Single fcz file downloads he fcz file can either be saved to the remote computer or device or AlohaView software can open it automatically Zip file downloads Save the zip file to the remote computer or device Then extract the file to the desired location using WinZip or other extraction program NOTES To open fcz files AlphaView Stand Alone software version 3 3 or higher must be installed on the remote computer The remote computer must be Windows based to run AlphaView software FluorChem E System User Guide page 108 Chapter 9 Networking and Remote Access Opening Image Files in AlohaView Software Stored image files can be opened directly in AlohaView software for post acquisition image analysis NOTES To open fcz files AlohaView Stand Alone software version 3 3 or higher must be installed on the remote computer AlphaView software is only available in a Windows compatible version Both fcz and png files can be opened in AlphaView software Opening fcz Files Opening files during download Foll
85. s in this page select the Edit button This will display the Trash Edit page FluorChem E System User Guide page 82 Chapter 7 Managing Stored Images Delete Copy Move Button Button Button ee Cancel 3 Images Selected TI Delete ie Mow Button Select All Button Last Week _ Paging Image List Buttons A000 PR EE POS Lee POA SNE AD 0 ETAT 1S PAA S AAT LAA PAn AF FOUTS EDA O PS TTS ON STL 0 A T Two Weeks Ago ETAM a N A 101 1 WTSinni AOtiDn nI AMESA OED A P a Le A With the exception of how images are deleted and the Move button the Trash Edit page options are the same as the Album Edit page Please refer to Editing the Image Album on page 79 for details on how to use this page Deleting Images from the Trash Image files can be deleted from the Trash Edit page in the same way that they are deleted in the Album Edit page However images deleted from the Trash Edit page are permanently deleted Moving Images back to the Image Album To move one or multiple images back to the Image Album select the image s in the Image List then select the Move button The following dialog box will display Move selected images to Album FluorChem E System User Guide Backup and Restore page 83 Select the Move button in the dialog box The image file s will be moved out of the Trash and back into the Image Album Exiting Trash Selection To exit the Trash Edit page select the C
86. t of the image acquisition process FluorChem E System User Guide Flat Field Calibration page 99 IMPORTANT Flat field calibration is performed on the FluorChem E prior to system shipment Recalibration is periodically recommended depending on variations in illumination across the field of view Please contact ProteinSimple Technical Support for further assistance 1 Goto the System Settings Tab and select the Flat button The following dialog box will display Create Flat Field Masters Flat Field Masters created 06 Apr 2010 You will be asked to place a screen on the light box Multiple images of each screen will be taken and used to generate flat field masters Estimated duration 1 30 hours 2 Select Start This will prompt the following message Create Flat Field Masters Place the white screen on the light box and close the door maiii If UV white light flat field calibration is not required select Skip to proceed to the completion step Oth erwise place the white screen used for colorimetric imaging on the UV transilluminator Close the system door and select Continue The following status screen will display FluorChem E System User Guide page 100 Chapter 8 System Settings g T i 7 Create Flat Field Masters Taking the white and UV flat field images Started 09 31 AM 15 Feb 2011 Image 1 32 Exposure 200ms Expected completion 10 01 AM 15 Feb 2011 The sta
87. the system through a local network FluorChem E System User Guide Overview of FluorChem E System Components page 17 USB Ports The FluorChem E system has two side and one top USB port These ports can be used with USB flash drives for copying image files or to connect directly to a compatible USB thermal printer System Label The FluorChem E system label is located on the rear panel of the instrument and includes the following infor mation ProteinSimple location system model power and fuse requirements serial number and certification markings An additional serial number label is located on the inside of the FluorChem E system door proteinsimple Santa Clara CA USA Model FluorChem E Made in USA 1O00VAC 230VAC 50 60Hz 345 Watts H FUSET6 3AH 250V _ s n System Power The power components consist of the main power switch power entry and fuse They are located on the system s rear panel IWARNING Only use the power supply cord provided with the instrument If the power supply cord is damaged please contact ProteinSimple Technical Support WARNING Only use fuses rated for 250 V 6A IWARNING SHOCK HAZARD Disconnect power cord from the instrument power inlet to disconnect power to the instrument Sample Screens The FluorChem E system is provided with three sample screens for imaging a wide range of applications Sam ple screens are application specific and should be used as follows Black
88. toggle the Zoom button to Off FluorChem E System User Guide Image Zoom Zoom Off 22 of 28 P dont com mer Q lt q 200 Zoom 22 of 28 Downou com mont R 400 Zoom 22 of 28 ori tn mont FluorChem E System User Guide page 65 page 66 Chapter 6 Image Review Viewing Pixel Values Pixel signal intensity can be determined using the Pixel Viewer option To view pixel values select the pixel of interest on the image The Pixel Viewer will display pixel values as well as a horizontal bar graph indicating the degree of saturation for each channel Saturation is indicated by a value of 65 636 NOTE The saturation value for stacked channels those that use the same light source increases by 65 636 per additional stacked channel Close Button Pixel values for other locations on the image can be viewed by selecting another pixel of interest To exit the Pixel Viewer select the Close button in the upper right corner of the values box FluorChem E System User Guide Browsing Images page 67 NOTES Pixel values and degree of saturation depend on the channel image data currently displayed Channels that are hidden from view are not included in pixel value calculations Calculated pixel values are based on 16 bit image files and are unaffected by image adjustment settings Browsing Images The Older and Newer buttons allow users to browse images saved on FluorChem E in the Re
89. view Screen Selecting either button will show images as they are currently dis played in the Images Screen q Older Newer p FluorChem E System File Information FluorChem E system files have a fcz file extension If a Title was entered in the Image Info box this will also be used for the file name For example if the title is LOD experiment the file name will be LOD experiment fcz Ifa title is not entered the default file name will include the date and time of acquisition An example default file name would be 2010 06 16_09 22 15 fcz The fcz file format is a proprietary zipped file format that can be opened directly in AlphaView software for post acquisition data quantitation Each file contains image acquisition and user entered information as wellas 16 bit ong files of each channel image and an 8 bit per channel ong file This 8 bit per channel ong file is the overlaid multi channel image or single channel image based on the selection in the Exposures page Once opened in AlphaView software Fluorchem E system cz files can be converted to tif or jpeg files Copying Printing and Deleting Images Copying Images To copy an image select the Copy button Insert a USB flash drive into one of the three USB ports on the FluorChem E system When the drive is detected the following message is displayed FluorChem E System User Guide page 68 Chapter 6 Image Review Copy to USB flash drive x Detected USB drive
90. ystem User Guide page 87 Chapter 8 System Settings Chapter Overview Overview of the System Settings Screen Changing System Information Changing Date and Time Settings Changing Filter Settings Updating the Software Saving Custom Protocols System Self Test Creating Dark Masters Flat Field Calibration Changing Focus Settings System Log FluorChem E System User Guide page 88 Chapter 8 System Settings Overview of the System Settings Screen Selecting the Settings Tab will display the System Settings Screen This screen contains system information and allows users to change the system name date and time and focus settings as well as install software updates and run system tests Dark masters flat field calibration and system log files can also be generated System Settings Tab lt a Software and Darkroom Review images y System z E7 imag Firmware Information l FluorChem E FE0232 ft rae lt _________ Versions Laboratory l 5 FE0232 htu el il Network with DHCP lt A Network pre TR Information Optics 00 18 7d Of 25 bs Temperature gt soc Name Time Filters Update Test SettingsAccess Buttons mas Flat Focus Protocols Log Backup System Information FluorChem E system information is located in the upper left corner of the System Settings Screen FluorChem E FE0232 Laboratory SN F EQ 3 al ttp A Fi fe 2 3 2 IOC al The system serial number is located under the inst

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