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PerkinElmer 2030

1. Ready Preset Protocol editing This tab allows you to select the protocol number and name The number can be selected from a drop down list of available numbers The protocol name has to be typed in In the case of a factory pre set protocol you cannot change the number Notes or additional comments can be connected with the protocol These appear at the bottom of the Explorer window Under this tab you can also see information about the protocol when it was created when it was edited most recently by whom it was created and edited and when an assay was last run with it The user names used come from the identifier given when Windows was booted Pre set protocols are created in the factory 53 Protocol editing In the enhanced security mode the first user name is the system user name and the second is the PerkinElmer 2030 user name User names are separated with a slash Measurement The measurement tab specifies the measurement sequence to be used in the protocol iB Copy of Fluorescein High Count 0 15 Protocol editor Y O E Eile Help ID Samples Measurement Outputs and Events Measurement mode C By plate By wells C By strips Operations Operation properties E 010 4190 3 6 4 Ready Note If the protocol being edited is pre set the measurement sequence cannot be changed Measurement mode This fiel
2. EEN dee EEEa a a Oa ERE r ERE EA SEa REESS 21 Fe Mperature vis oes edu Auge EE E Eae o Ep Einen neh ent 23 Barcode iia 24 Plate orientation ia 24 Starting operation with the Start button 25 Starting operation with the Start Wizard none 29 Starting operation and using a protocol number barcode nen 34 Stacker operatio a e a ib ios ita 36 Starting operation no protocol number barcode nenn 36 Starting operation and using a protocol number barcode nenne 38 Epding OPEL see eri ea ia aE e EEN 39 Stop button E SE EEEE TTET 39 Button on PerkinElmer 2020 40 Light on the button 2 2a e eebe 40 Protocol editing sisssssssssssveisssssssssivessisssoesssseosssas vosvos N 41 Lt Oe A3 Explorer folders a 44 Explorer VES A ias 44 Explorer Icons IAE nee EA SEA deidad 47 Protocol seleciona E E EE A 48 Protocol editor s A REO 49 Contents Samples r 2 02 H 0 ee Aeddi E e E E tee ceuviesd 50 Plate repeat parameters ioris onnies eke ireu Ee SEENEN dE 51 Me surement heisht 2 2 2 2a222220 lerne Ri 51 Plate type id 51 Temperature level check otitis 52 Di A taa 53 NES O 54 Me s rement oc eege EES Shamnatacbeetveds 54 Measurement operanons nono nocn nono R E cnn cnn cnn nnnn cnn R ERS 55 Down Butto ae stehen daa iia 55 Up button 2 335 5 site EE asias 55 Delete tton cuida ar Laser up un nn 55 Dispense DUO 55 KOENEN 56 Shake Dutton iconos 8 EES 56 Label 4er ENEE see AER NEE ee 56 Kinetics button 2 sR H R
3. Protocol last edited by protocol last edited Absorbance 280 1 03 200 1 08 Photometry Mallas 6 16 2004 Maller 6 16 2004 46 Protocol editing This icon is for List View In this view the assay results if any of an highlighted protocol is displayed in a single active window Porkinfimer 2030 Explorer Que Ele CN To Wem ep er rege NA Explorer icons Factory pre set protocols and groups have a lock symbol to show they cannot be changed User made protocols and groups do not include the lock because the user can change them See the examples below 8 The icon for a factory pre set folder The icon for a factory pre set protocol The icon for a user made folder The icon for a user made protocol 47 Protocol editing Protocol selection You can Open copy paste start or create a protocol delete a user made protocol plus its associated results In the enhanced security mode you have to belong to the Operator user group to start a protocol To open an existing protocol you can do one of the following a Move the mouse cursor onto it and then click the left mouse button to select it then click the File menu and click Open b Move the cursor on to it click the right mouse button and select Open from the menu that appears c Double click the mouse when the cursor is on the icon At the bottom of the window on the status line the protocol ID will appear if d
4. This is the name by which the label is identified A password can be associated with it to prevent parameters being changed by an unauthorized person CW Lamp energy In fluorescence polarization constant voltage mode is used and the light source is driven with high power The software corrects for changes in energy of the CW lamp to ensure that 88 Tools menu results are calculated for a constant excitation energy For fluorescence polarization you should always use maximum power 65535 CW Lamp filter You can select the CW lamp excitation filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory pre set CW lamp filter names begin with a letter indicating the technology for which they are to be used F is for fluorometry After the letter comes the wavelength Polarizer aperture There are two sizes for the aperture of the S plane polarizer in the excitation light path You can select by option buttons whether this is Small for high density plates or Normal 96 or 384 well plates Emission filter You can select the emission filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory preset emission filter names begin with a letter indicating the technology for which they are to be used F is for fluorometry After the letter comes the wavelength Emission aperture In front
5. A sample plate can then be loaded and measured using the protocol set for it Other parameters affecting the whole system e g labels filters etc can be set in the Tools menu 14 Operating PerkinElmer 2030 16 Operating PerkinElmer 2030 Starting up PerkinElmer 2030 Without enhanced security mode When you boot the system the following window appears The main window has three tabs Live Display Kinetic Chart and Temperature PerkinElmer 2030 Manager ll File Instrument Tools Help P r ale Biz ole Live display Kinetic Chart Temperature Protocol Absorbance 450 0 15 2 gt 1 JE JS HI Ss e a 12 Seale Logarithm a Linear Extra High y N 5i 10000000 6 1000000 D 100000 E 10000 1000 F 100 o _ 10 H 0 Measurement information Not available yet For Help press Fi Temp C MA DEMO With enhanced security mode When the Enhanced Security mode is installed the login dialogue appears after you have started PerkinElmer 2030 17 Operating PerkinElmer 2030 PerkinElmer 2030 Manager Fie Instrument Tools Help Sei ple de Live display Kinetic Chart Temperate Protocol Absorbance 450 0 12 7 Bl sl 12 1 2 3 4 GES 7 8 H 10 11 Linea 8 PerkinElmer Login SB Enter your user name and password User name 1000000 Bassword 100000 Ca ee em H Mesturemerk infcemation Not av
6. Meas warning page Eu Sm norm 110 Meas warning page LANCE norm 115 165 Index Miscellaneous 73 Miscellaneous settings window buttons 125 MultiCalc 143 144 MultiCalc assay protocols 143 144 N Name of Instrument 127 Name of plate 126 Needle maintenance 153 New user 155 Nickname of instrument 127 Normalization 107 112 Norm confirmation page Eu Sm norm 111 Norm confirma page LANCE norm 116 Normalization wizard dual label 107 Notes Result Viewer 69 Number of well columns Plate type 126 Number of well rows Plate type 126 O Operator 154 Options 73 127 Options Protocol Explorer Tools 61 Options installed 130 Orientation of strips 127 Output 11 Outputs Protocol editor 58 Outputs and Events tab 49 58 Outputs tab 60 File output 59 Printer output 58 Overview 11 P Padding keywords 60 Password 77 81 84 86 88 91 Password control 156 Perform on first plate repeat only 56 Performance degradation 139 Photometry 84 Index Plate Add 50 Delete 50 Duplicate 50 Plate dimension wizard 73 Plate dimensions fine tuning 119 Plate dimensions Plate type 127 Plate format 58 Plate heating 23 Plate heating Plate parameters 52 Plate height 126 Plate map options 129 Plate map results picture 68 Plate name 126 Plate name Plate type 126 Plate orientation picture 146 Plate repeat parameters Plate parameters 51 Plate type
7. NUNC 136528 Terasaki 72 Wallac 1450 514 Isoplate Wallac 384 White AAAND 0002 NUNC 163118 Terasaki 60 Costar 384 OptiPlate 96 OptiPlate 384 ProxiPlate 96 ProxiPlate 384 ViewPlate 96 ViewPlate 384 N A N A N A N A N A N A N A N A N A N A N A N A N A N A N A N A N A N A 108 Tools menu Preparing the plate PerkinElmer 2030 EuSm Dual label normalization Wizard Prepare a plate so that strip A is filled with blank samples and strip C is filled with europium samples and all others strips are left empty Load the plate into the instrument Click Next to start the measurement of the plate The wizard explains what samples you should put in the plate and where you should put them Plates must have 3 or more strips maximum of 8 with 1 12 wells per strip The strips must be horizontal e g Al to A12 etc Strip A should be full of blank samples Strip B should be empty and Strip C should be full of europium samples 109 Tools menu Measurement status page PerkinElmer 2030 EuSm Dual label normalization Wizard The measurement has now been started Measurement status Assay is running Measurement progress Measurement in progress please wait This is shown during the actual normalization measurement and it gives you feedback about the measurement The measurement status pane shows the current st
8. close the cover and switch power on 104 Tools menu Changing filters in the software Note If you change a filter in the software you must also change it physically in the filter slide or wheel When you select Properties a picture of the selected slide appears with the filters that are loaded in it marked in red or black as shown previously Those that are in red are factory pre set The excitation filter slide is in the form of a wheel and the emission filter slide is a strip To move a filter from the list to a place on the slide move the cursor to the filter name on the list drag it keeping the mouse button pressed down until it is over the empty slide position that you want it to occupy then release the mouse button The name of the filter will appear in black in the slide position To move a filter from one position to another click on it and drag it to the empty place you want it to occupy To remove a filter from the slide click on it and drag it back to anywhere in the area of the Available filters list then release the mouse button 105 Tools menu Filter properties The filters used in the instrument have the properties shown in the table below The following abbreviations are used CWL Centre wavelength BW Bandwidth Full Width Half Maximum Tpk Maximum transmittance at the peak wavelength Filter type Tr fluorescence excitation Tr fluorescence excitation Europium chelate emission LAN
9. 49 52 Plate type Miscellaneous settings 125 Plate type selection Eu Sm norm 107 Plate types and Well names 124 Plate types for stackers 145 Polarizer aperture 89 Preparing the plate Eu Sm norm 109 Preparing the plate LANCE norm 113 Preview print 61 Print Protocol editor 61 Print Result Viewer 70 Print preview Protocol editor 61 Print setup Protocol editor 61 Printer export 70 Processing error 139 Product number 127 Protocol Result Viewer 69 Protocol editor 49 Protocol ID 48 Protocol name 77 86 88 91 Protocols Number of Max 40 49 Pumps 147 166 R Reading time Photometry 85 Repeat for plate 51 Result viewer Error 70 Export 70 Notes 69 Plate map 68 Preview 70 Print 70 Protocol 69 Result Viewer 65 66 67 69 70 Results export 70 Results of latest assay run 65 Results picture 68 Results viewer File menu 66 List 67 Plate menu 66 View menu 66 Results viewing 65 Routine level 74 Rows of wells 126 S Samples protocol editor 50 Samples tab 49 50 Save Protocol editor 61 Scanning button Measurement tab 57 Second Emission filter Luminescence 87 Second meas parameters Fluorometry 83 Second meas parameters LANCE 94 Second measurement parameters TRF 80 Select where output goes Protocol editor 58 Serial number 127 Service 154 Service level 74 127 Shake button Measurement tab 56 Shaker Path 56 Slide emission filte
10. 86 89 93 Archive 159 Archive old results 159 Archive old results 159 Audit Trail 157 Archive button 158 Export button 158 Protocol button 158 Auditor 154 Automatic logout 135 Barcode 24 31 34 35 36 38 40 60 128 129 Barcode options 24 128 Barcode reader option 24 Beam size LANCE 94 Beam size Time resolved fluorometry 80 Button light 40 Button on PerkinElmer 2030 40 Buttons Measurement tab 55 Buttons in the filters window 96 Buttons in the Labels window 75 Buttons in the Misc settings window 125 163 Index C Change password 18 Change the CW Lamp 104 Changing the CW Lamp filter wheel 104 Changing the emission filter slide 101 Column width changing 65 Columns of wells 126 Constant voltage mode 82 Control modes Constant voltage mode 82 For fast kinetics 82 For fluorescence polarization 82 Stabilized energy mode 82 Counter position Fluorometry 83 Counting parameters LANCE 93 Counting parameters TRF 79 CW_Lamp control mode 82 CW Lamp changing 104 CW Lamp energy FP 89 CW Lamp energy Fluorometry 81 CW Lamp filter wheel 102 CW Lamp filters 82 85 89 98 D Define labels 75 Delay button Measurement tab 56 Delete button Measurement tab 55 Demo 19 Demo mode 127 128 Dimensions of a plate 127 Dispense button Measurement tab 55 Dispenser information 147 Dispenser maintenance 73 150 153 Needles 153 Down button Me
11. Flash Energy Area selects a 91 Tools menu basic capacitor of 10 nF The High Flash Energy Area selects a second 10 nF capacitor in the lamp s discharge circuit thus doubling the energy If you select High you will reduce the counting time number of flashes but the counts will remain about the same Flash Energy Level The Flash Energy Level sets the voltage of the discharge capacitor the range is from 1 to 255 corresponding to 500 to 800 volts Note that although the lamp input energy depends on the squared power of the discharge voltage the output optical power is not linearly dependent on the input energy By changing this parameter you will fine tune the Flash Energy If you increase the number you will reduce counting time but counts from your sample will remain about the same Excitation filter The non changeable filter in the flash excitation path D340 has a maximum transmittance at 340 nm and half bandwidth of about 35 nm The other two filters are only used for UV absorbance Light exposure area A reference circuit monitors the total excitation energy When the selected total amount of energy has been reached the flashes stop There are three different light exposure time selections short medium and long any one of which can be selected The capacitor selected determines the total excitation energy in the same proportions as the capacitor numbers within component tolerances This means t
12. Fluorometry DELFI amp E EI Fluorescence polarization Time Resolved Fluorometry LANCE 2 2 lt Back Cancel He Preparing the plate PerkinElmer 2030 Plate Dimension Wizard Prepare a plate so that the wells in each comer are filled with appropriate liquid depending on the label used and leave all other wells empty For example the comer wells of the 1536 plate are in the following positions Well 1 A1 first well of the first row Well 2 A48 last well of the first row Well 3 AF1 first well of the last row Well 4 AF48 last well of the last row Size of the scanned area per well 1 40 mm Load the plate into the instrument Click Next to start the measurement of the plate coca mm Tools menu Prepare the plate by dispensing appropriately labelled sample to each of the four corner wells of the plate All the other wells should be empty Specify the area of the well to be scanned The example shows the default value for a 1536 well plate 121 Tools menu When the plate has been prepared load it and click Next 7 PerkinElmer 2030 Plate Dimension Wizard The measurement has now been started Measurement status besen is running Well scanning progress Well 1 wei wel wei Measurement in progress please wait Help After scanning is completed the Next
13. Plate selection Note This step requires that there is at least one user defined plate E PerkinElmer 2030 Plate Dimension Wizard Select the plate type which dimensions are to be adjusted Plate Name W Horiz Verti Hor Ver My OptiPlate 96 8x12 14 380 11 240 9 000 9 000 coca to Select the plate whose dimensions you want to fine tune Remember that this should be the plate type with the name you chose specifically for the technology to be used Note that you must click in the Plate name field to select the plate When you have done this Next will be enabled Click it to go to the next dialogue Technology selection You must select the technology to be used to count the plate Note The options that appear will depend on what is installed in the instrument Open the folder and select the actual protocol for which the plate dimensions are to be fine tuned The Next button will then be enabled and you can click it to go to the next dialogue of the wizard 120 PerkinElmer 2030 Plate Dimension Wizard Select the label that is used in the scanning The label should use the same technology that is used in actual measurements with the plate Lg Fluorometry Fluorescein High Count D Del Fluorescein D Del Umbelliferone D Del Fluorescein 0 15 Bottom Fluorescein High Count 0 15 Umbellferone 0 15 Luminometry Photometry la ta Time Resolved
14. a barcode is used it should also be on the side that enters the reader first PerkinElmer 2030 Start Wizard Protocol DELFIA cAMP kit Notes cAMP 050308 SQA measure lt Back Next Cancel u Please insert first plate to be measured in instrument and press Finish button to start to Click Finish to start measurement See the Live display during measurement 31 Operating PerkinElmer 2030 When the plate has been measured remove it load the next plate and briefly press the button on the reader The steps when creating a new protocol are Introduction to the wizard You can select the introduction not to appear next time if you want Create new protocol select the location for the new protocol and type in the name you want to use PerkinElmer 2030 Start Wizard Y Use existing protocol Create new protocol Test protocol 18 Protocols 1 Lei PerkinElmer L Assay examples L Users EQ User Kinetic Y User 2 DI User 3 User 4 User 5 2 User6 2 User A User 8 User 9 Y User 10 Y Extemal lt Back Cancel Finish Plate selection select the plate you want to use with new protocol 32 Operating PerkinElmer 2030 PerkinElmer 2030 Start Wizard A Please select the plate for the new protocol and click Next to continue Name Rows Col Origin Origin Generic 8x
15. absorbance Light Exposure Time A reference circuit monitors the total excitation energy When the selected total amount of energy has been reached the flashes stop There are three different light exposure time selections short medium and long any one of which can be selected The capacitor selected determines the total excitation energy in the same proportions as the capacitor numbers within component tolerances This means that selecting medium instead of short approximately doubles the excitation energy for one measurement i e the number of flashes for one measurement doubles hence the time used for one measurement doubles and the number of counts doubles Selecting long instead of short triples the excitation energy the measurement time and the number of counts The number of flashes can be seen from the result viewer by selecting View all result data Note Each flash contributes to the total excitation energy Flashes keep on occurring until the required total excitation energy is achieved However because the energy comes in discrete flashes the last flash will cause the total excitation energy to exceed the required amount by a maximum of not more than the energy in one flash This will not be significant if there are many flashes but could be significant if there are only a few flashes E g in standard measurements the amount of flashes is about 1000 the variation in the total excitation energy is thus 1 1000 or 0 1 If the a
16. all result data only when List is selected Toolbar Status bar Plate first plate previous plate next plate last plate all with corresponding buttons These buttons or menu items that allow you to step between plates only appear when the plate mode is selected Help information about Result Viewer 66 Result viewing List E Assay 7 3 6 2008 1 26 33 PM PerkinElmer 2030 Result Viewer Woe File view Plate Help SIAE 1 Plate Repeat Well Type Time Europium 2 1 1 401 M 00 00 44 00 388603 3 00 00 44 88 785696 4 00 00 45 76 127892 5 00 00 46 564 254727 6 00 00 47 52 707051 7 00 00 48 40 577255 8 00 00 49 28 463471 3 00 00 50 16 204271 10 00 00 51 04 698682 11 00 00 51 92 874765 12 00 00 52 80 364583 13 00 00 53 68 218169 14 00 00 54 56 161519 15 00 00 55 44 84083 16 00 00 56 32 818327 17 00 00 57 20 305546 18 00 00 58 08 551815 19 00 00 58 96 771013 20 00 00 59 84 549458 21 00 01 00 72 371351 2 00 01 01 60 552178 23 00 01 02 48 309806 24 00 01 03 36 932775 25 00 01 04 24 452897 26 00 01 05 12 931113 27 00 01 06 00 189557 28 00 01 08 88 473683 29 00 01 07 76 571484 30 00 01 08 64 656232 n Ai af rh List Plates 1 1 Plate Paget Protocol A Errors A Notes Lett K Ready Errors may have occurred in measurement Check results The following information appears Plate ID Number of repeats Well number and Sample type abbreviation e g M Measured In addition there are pairs
17. display the folder information as a list just showing folder names or with additional details The white area in the centre of the window is where the folders appear when you start to browse to find the path 70 Tools menu 71 72 Tools menu Tools menu Note These operations can affect the whole system and should only be undertaken by someone qualified to do them There are up to fifteen items in this menu depending on the options installed Explorer for protocol editing and result viewing see pages 43 and 65 Start Wizard for starting when you first need to define plates and wells see page 29 Latest Protocol run opens the Protocol editor with the protocol that was used in the latest assay run Latest Assay results the result viewer is started and the latest results shown see page 65 View Picklist protocol opens the Protocol editor with the protocol that is active in the protocol picklist Labels for defining labels used in measurements Filters for defining filter types Plates and Wells settings including plate types and well names EuSm dual label normalization wizard requires TR FIA to be installed LANCE normalization wizard requires LANCE Plate dimension wizard for fine tuning the plate position when measuring high density plates Dispenser maintenance described in Appendix 3 Miscellaneous settings User level does not appear in the enhanced security mode sel
18. is different from the current one all the plates in the plate map will be 51 Protocol editing deleted and one plate of the selected type will appear Make sure you select the plate type before defining the plate map to avoid losing your settings Plate types are read from the PerkinElmer 2030 database when the Protocol Editor is started so it will not recognize plate types added while the Protocol Editor is open Note The generic 8 x 12 plate selection does not check the height of the plate before a measurement Note If the type you selected has a different number of wells than the current one all the previous plates will be deleted and one plate of the selected type will appear Temperature level checking This setting allows you to define what the target temperature should be when a plate is measured Note Setting this option will not activate the plate heating automatically but it will cause a warning to appear if the temperature differs significantly from what has been specified in the protocol 52 ID Fluorescein High Count 0 15 Protocol editor File Help He Identification amples Measurement Outputs and Events Protocol name Protocol number Notes Protocol information Created by Creation date Last edited by Last edited Last run Run count Wallac 1 31 2000 12 00 00 AM Wallac 1 31 2000 12 00 00 AM 3 6 2008 9 49 34 AM 1
19. of columns The number of pairs depends on what you have set in protocol editing parameters The first column of each pair is the measurement time and the other the result value You can change column widths to get more information In the case of high density plates results can be split over several tabs View all result data This item in the View menu is only active if List view is selected It causes an extra column to appear in the result list giving the following extra information Time resolved fluorometry number of flashes Fluorescence measurement time Luminometry counting time Photometry A D converter value and the number of readings 67 Result viewing Plate map This plate map shows you the results arranged from top to bottom in the order they appeared in the list from left to right See List format for the alternative way of viewing results Assay 7 3 6 2008 1 26 33 PM PerkinElmer 2030 Result Viewer Loe File View Plate Help Sol ale ejeje 41 gt gt i hu 1 A A A A 7 CA TH 12 Plate Repeat Endtime Start temp Endtemp Barcode El 1 11 43 14 PM NA NA 1 Europium Counts 2227 A 388603 289503 830744 995830 901625 56165 591636 452564 434406 590458 716961 486161 785696 197020 784820 624387 616290 202338 835367 136563 344552 798990 866547 118344 127892 240799 255332 386458 924406 54474 375007 317299 885128 920297 920387 777992 254727 978216 64686 73829
20. protocol barcode option is used Condition Barcode none or plate Barcode for protocol number number Not measuring no assay Off Blinking Not measuring waiting next Blinking Blinking plate Measuring loading or On On unloading 40 Protocol editing 41 42 Protocol editing Explorer In the enhanced security mode Explorer is enabled only for users who belong to the Editor user group Through the Explorer you can access the Protocol Editor and the Result Viewer Click the Explorer icon to open the Explorer Elmer 2030 Manager strument Tools Help ale ple le Q PerkinElmer 2030 Explorer Im x im lg aja Errors Begin date 8 Protocols 8 PerkinElmer da Assay examples Ca Luminescence Assays BB atPlite ATPlite 1step britelite plus easylite Kinase steadylite plus Lei Bradford Protein Assays C DELFIA Assays 8 TruPoint Assays L LANCE Assays Ca Aequorin Assays L Users CA External Protocol number N A 43 Protocol editing Explorer folders In the PerkinElmer folder there are as a default folders named according to the technologies installed In each of these folders there are protocols using the appropriate technology There are also Example assays folder including example protocols for some typical applications On the same level as the previous folders there are two other folders Users and External External shows protocols cr
21. r Upon assay completion a new assay measurem nt is automatically started using the same protocol 103 Tools menu Tools menu Physically changing the CW Lamp filter wheel If you want to change a filter in the CW Lamp filter wheel or you want to install wheel B then you must open the cover and lift out the filter wheel as shown in the picture Either change the filters you want or take a new filter wheel B Replace the filter wheel and make sure you close the cover before starting a measurement HOW TO REPLACE THE WHEEL AND THE LAMP ATTENTION KEEP THE LID CLOSED Oo LIFT THE LAMP VERTICALLY OFF FROM THE CLIPS HOLDING IT LIFT THE FILTER WHEEL VERTICALLY OFF FROM ITS SUPPORT REPLACE THE WHEEL AND CLOSE THE CLIP Ea PULL THE LAMP OFF FROM CAUTION THE LAMP SOCKET DO NOT TOUCH THE OPTICAL COMPONENTS AND THE LAMP BULB OR THE INSIDE OF THE WARNING LAMP REFLECTOR WITH BARE FINGERS LAMP AND OPTICS MAY BE HOT CODE 1420 401 Changing the CW Lamp If the CW Lamp needs replacing lift the cover to get access to the lamp Make sure the lamp is switched off and give the lamp chance to cool before removing it and replacing it Follow the instructions on the inside of the cover see the picture on the next page You will probably find it easier to use your left hand to remove the lamp from the clips holding it When you have replaced the lamp
22. select one of more of these parameters two measurements are made the first with the normal settings and the second with these new settings The change is automatic 83 Tools menu Photometry A reference measurement is made before the plate is moved to the measurement position This enables the absorbance to be calculated Photometry Label Properties Name Bbsorbance 4051 0 tt Password Last Edited Nouser 2 15 2008 1 39 20 PM Parameters Absorbance Mode Ze Visible C UV Flash Lamp Filter E fs c CW Lamp Filter P405 Slot 41 v Excitation Aperture C Small Normal Large Reading Time 1 0 1 600 s Second Measurement Da Lamp Filter P595 No Slot v Cancel Help The parameters that can be set for photometry are the following Label name This is the name by which the label is identified A password can be associated with it to prevent parameters being changed by an unauthorized person Absorbance mode Select whether you want visible light or ultraviolet absorbance Flash Lamp Filter These option buttons are only enabled if UV absorbance mode is selected Select the wavelength of the filter you want The filter will ensure that only UV light of the selected wavelength is used for the absorbance measurement The UV photometry filters P280 and P260 have their transmittance peaks at 280 nm and 260 nm respectively 84 Tools menu CW Lamp filters You can select the CW lamp filter
23. slides in the currently selected tab are displayed Names of user defined filters can be freely defined by the user but names of factory preset filters cannot be changed There are four buttons at the bottom of the window Add add a new item to the list available e g a new filter not available for slides Give the name and edit the properties It will appear with a new icon showing it is different from a default item Copy select an item to activate this not available for slides Make a copy of an item It will appear with a new icon showing it is different from a default item and the name will be preceded by the words Copy of You can give it a different name and edit other parameters by selecting Properties Remove select a user created item to activate this not available for slides Remove the selected item from the list You cannot remove a pre set item from the list nor can you remove a filter that has been assigned a position in a slide This command requires confirmation because you cannot undo Remove Properties select an item to activate this View the properties of the selected item If the item is user defined then you can edit the properties but if it is a factory pre set you cannot edit them 96 Tools menu Emission filters Filters Emission Fitters Cw Lamp Filters Fiter Slides D545 D572 D615 D642 D665 F460 F535 No filter Help If you select proper
24. the operation currently selected in the measurement operations list is not the first operation in the list Delete button This deletes the operation selected in the operation list This button is enabled when there is an operation selected in the measurement operations list Dispense button Clicking this causes a dialogue to appear that lets you specify the parameters for a dispense operation This button is enabled when the dimensions of the plate type used in the protocol are 8x12 or less You can select the number of injectors to be used from 1 to 4 depending on how many are installed the volume to be dispensed in microlitres A slide bar helps you to select this volume 350 microlitres is the maximum the speed from 1 to 5 where 5 is the fastest and 4 the default 55 Protocol editing the increment using a slide bar with a range from plus 100 microlitres to minus 100 microlitres this is the volume added to or subtracted from the previous volume dispensed before the next dispensing the number of replicates from 1 to 3 the injection mode You can either aspirate and then dispense the same amount aspVol dispVol or you can fill the whole syringe and then dispense several times aspV ol syringeVol The former is more accurate but the latter is faster At the bottom of the dialogue is a check box Perform on first plate repeat only If you only want dispensing for the first of the plate repeats check this box If
25. to be greater than or equal to the Maximum password age The minimum is day and the maximum is 999 days Minimum password length defines the minimum number of characters that must be used in the password The default number of characters that must be used in the password is 6 The minimum is 1 and the maximum is 16 characters Password uniqueness defines the number of new passwords that must be used before a previous password can be reused For example if the first password is security and Number of passwords to remember is set to 3 entries users must use 3 other passwords in addition to their current password before they are able to reuse security as their password The minimum is 1 and the maximum is 24 Click Account Lockout to set the lockout properties The Account Lockout dialogue will appear Lockout defines the number of failed login attempts that are allowed before a user is locked out of PerkinElmer 2030 Manager For example if Number of failed logins allowed before lockout is set to 5 failed login attempts the user is allowed 5 failed attempts to log in On the 5th failed attempt they are locked out until the Administrator allows them access again Permanent or for a specified period of time Duration The default is lockout after 5 failed login attempts The minimum number of failed login attempts before a user is locked out is 1 The maximum number of allowed failed login attempts before a user is locked o
26. when the end of the plate map definitions is reached for a protocol There are three possibilities Assay ends after last plate in the plate map no more plates will be measured until a new start instruction is given or a new barcode read If there are more plates an error message will appear Additional plates possible latest plate map is used further plates will be measured using the last defined plate map Additional plates possible all wells will be measured further plates will be measured with no plate map taken into consideration i e every well will be measured There is also a check box Upon assay completion a new assay measurement is automatically started using the same protocol This check box is only enabled if the Barcode stored to results option has been selected If you are using stackers then measurement of the next plate with the previously used protocol will begin automatically This feature is useful for automatically producing a separate export file for each plate Define your protocol to have only one plate and select this feature Each plate will be run as a separate assay producing its own export file The external software can then process these files as from the same assay because each will have been produced by the same protocol If you are not using stackers the instrument waits until the button on the instrument is pressed and then begins measuring the current plate with the previousl
27. which you can give the number of horizontal steps the number of vertical steps and the point displacement i e the distance between each point on the well where the measurement is made The total number of points can be a maximum of 100 The distribution of points can be square or rounded You also need to select the label by clicking the Label button and then selecting from the label dialogue The scanning button is enabled only when using well mode and there are no other labels in the measurement sequence However copying and pasting of the same scanning label is possible Injector position during measurement At the bottom of the Measurement tab are two check boxes for controlling the injector head These will only be enabled if you have selected the by wells mode You can select if you want the measuring head to be kept above the well during measurement This means that measurement can be started immediately after injection without waiting for the injector head to move aside If you check this box then a second check box is enabled where you can select if you want injecting and measurement to happen simultaneously This is for use in very fast kinetic measurements 57 Protocol editing Outputs and Events Copy of Fluorescein High Count 0 15 Protocol editor ul m Ele Help Hl ei ID Samples Measurement Outputs and Events JT Printer output Printer output options Include in ML output W Fil
28. 0 864 mm Vertical 0 315 mm Vertical 0 315 mm Deviation of well 3 7 Deviation of well 4 Horizontal 0 549 mm Horizontal 0 909 mm Vertical 0 531 mm Vertical 0 4866 mm r Location of well 41 r Distance between wells Horizontal 13 849 mm Horizontal 8 968 mm Vertical 10 925 mm Vertical 8 972 mm lt Back Cancel Help Numerical values are shown in the final window If you approve these click Finish to save the values and end the wizard If you feel the deviations are too large you can click Back to reposition the well centres You can alternatively click Cancel to end the wizard without saving the values Note For even better adjustment it is recommended that you repeat the scan with a smaller area selected e g for a 1536 well plate this could be 1 mm per well 123 Tools menu Plate types and Well names In the enhanced security mode this tool is enabled only for users who belong to the Editor user group Plate types and Well names Plate Types Well Names Instruments Generic 8x12 size pl Labsystems 8x12 DELFIA strip plate NUNC 8x12 DELFIA strip plate Costar 3512 3x4 wells Costar 3524 4x6 wells Falcon 3047 4x6 wells NUNC 384 NUNC 136528 Terasaki 72 ES Wallac 1450 514 Isoplate NUNC 24005 petridish 60x15 mm In the dialogue that appears when you click this item in the Tools menu there are three tabs for Plate types the available plate types are liste
29. 000 665nm This label is for fast decay Eu chelates for example Eu W1024 in a TR FRET assays e LANCE High Count 615 same as LANCE 615 but 1000 flashes Higher count rate e LANCE High Count 665 same as LANCE 665 but 1000 flashes Higher count rate e LANCE Long Decay 615 same as LANCE 615 but 3000 cycle time This label is for long decay Eu chelates for example Eu W8044 in a TR FRET assays 94 Tools menu e LANCE Long Decay 665 same as LANCE 665 but 3000 cycle time This label is for long decay Eu chelates for example Eu W8044 in a TR FRET assays e LANCE 545 same as LANCE Long Decay 615 but 545nm emission e LANCE 572 same as LANCE Long Decay 665 but 572nm emission e TruPoint 615 same as LANCE 615 but including 2 window 200 100 This is a label for Eu TRF Quenching assays e TruPoint 545 same as LANCE 615 but with 545 emission filter and 2 window 200 100 This is a label for Tb TRF Quenching assays Note 572nm and 545nm emission filters are not installed in the instrument by default 95 Tools menu Filters In the enhanced security mode this tool is enabled only for users who belong to the Editor user group In this window there are three tabs Emission Filters CW Lamp Filters i e continuous wave lamp excitation filters and Filter Slides including filter wheels Filter Slides allows you to specify the slides for emission and CW lamp filters The names of the filters or
30. 1000 x where CorrectedS S sample S buffer and CorrectedP P sample P buffer 90 LANCE Tools menu In LANCE homogenous time resolved fluorescence a flash lamp is used to illuminate the sample In the most common form of LANCE this excites the donor molecule which after a delay transfers the energy to the accepter molecule which then emits light Two labels must be defined one for the donor and the other for the accepter The picture shows the parameters that can be set for a LANCE label Lance Label Properties e Name LANCE 615 Password Last Edited No user 2 15 2008 1 42 21 PM Parameters Flash Energy Mode Low High Flash Energy Level 245 Excitation Filter Ze D340 C Optional Optional Light Exposure Time C Short Medium C Long Light Exp Ref Level 157 ee nn Emission Filter D615 Slot Al r Emission Aperture Ze Normal C Small Large Counting Delay 1 150 2 J0 0 65515 us Counting Window 1 100 2 0 0 65515 us Counting Cycle 1000 1000 65535 us Beam size e Normal C Narrow P Second Measurement Emission Filter y Cancel Help Label name This is the name by which the label is identified A password can be associated with it to prevent parameters being changed by an unauthorized person Flash Energy Mode There are two options for this parameter Low and High The energy of the flash lamp used is dependent on the discharge capacitor value The Low
31. 12 size plate 8 12 14 60 11 20 Labsystems 8x12 DELFIA strip plate 8 12 14 40 11 20 NUNC 8x12 DELFIA strip plate 8 12 14 80 11 20 Costar 3512 3x4 wells 3 4 24 85 16 70 Costar 3524 4x6 wells 4 6 17 55 13 75 Falcon 3047 4x6 wells 4 6 13 75 13 75 NUNC 384 16 24 12 00 9 00 NUNC 136528 Terasaki 72 6 12 30 46 26 65 Wallac 1450 514 Isoplate 8 12 14 30 11 20 NUNC 24005 petridish 60x15 mm 1 1 63 88 42 83 Wallac 384 White AAAND 0002 16 24 12 00 3 00 Helix 864 24 36 12 05 3 00 Costar 3516 2x3 2 3 24 90 23 15 NUNC 163118 Terasaki 6 10 34 80 2660 Greiner 1536 32 48 11 03 7 92 Costar 384 16 24 12 00 3 00 OptiPlate 96 8 12 14 38 11 24 OptiPlate 384 16 24 12 10 3 00 Di i gt Add measurement you can add a label by clicking the operation icon on the right side This opens the label editor where you can select copy or add the label to be used within the new protocol It is also possible to delete or change the sequence of measurement labels within the wizard page PerkinElmer 2030 Start Wizard d Please add the label s used for the new protocol and click Next to continue m Label s Europium slslalt lt Back Cancel Finish The rest of the pages of the wizard are similar to those that appear when you are using an existing protocol 33 Operating PerkinElmer 2030 Starting operation and using a protocol number barcode 1 When you lo
32. 2 639645 892017 117499 468911 153483 185025 884524 878179 707051 111577 980693 659918 112090 824490 859085 121638 14139 371864 169919 512024 577255 184481 794640 765091 763973 136261 202096 349084 981932 911988 670795 617680 463471 542479 210042 850655 494259 885672 470390 119885 442987 463562 90035 412894 204271 435343 118647 842377 60184 190887 226871 513021 250800 216387 692518 873859 698682 160613 463320 221976 83478 336998 537645 487370 797147 459846 892349 886730 874765 315245 267236 253519 560698 21994 537434 747416 274910 714756 982174 289926 364583 819143 638406 608254 921233 345247 208561 860021 857544 996374 786633 293098 218169 21390 711251 939331 497612 511359 228170 943108 408151 351380 170704 684844 161519 806121 773460 895522 996948 278928 521269 528974 347059 85200 153513 811408 84083 192730 600730 98313 681853 543355 788174 457519 764759 906550 932352 395099 818327 619402 983926 12024 978276 448697 338328 633633 393558 790591 621155 543083 305546 568765 24230 230557 760740 931597 652335 774517 541331 209770 1682880 599522 51815 620792 503806 832557 747325 417033 290923 145839 430841 746812 906368 900749 771013 394585 367423 259441 222943 10604 775696 709801 32085 371260 275424 95050 549458 908121 202821 588283 782403 68372 168136 368764 139252 478397 726291 164239 371351 494561 8
33. 2030 9010 01 March 2008 User manual PerkinElmer 2030 Software version 4 00 Sperkintiener 2030 Manager Woe Pis iaut Took mel se pa e Live play Kinetic Chat Temoeronan Prolocel Absorbance 450 1011 d BD ST a 5 DH D 10 D 2 G Le gge A f Lines a dc j O9 990 3903 WA 100A UE EU KA EE Ga KEELT EK EE Ka 9090 90 n oer Latest weng from Piste 1 repeat 1 label 1 wel GOT wet repeat 1 1 4004 Meauaing atray 2 using protocol Absorbance E 450 10 1 pP PerkinElmer precisely Foc Hebo cess F1 Momo C WA DEMO PerkinElmer Life and Analytical Sciences Wallac Oy P O Box 10 FIN 20101 Turku Finland Tel 358 2 2678111 Fax 358 2 2678357 Website www perkinelmer com Contents Contents Tradem rks 2 4 amp 5 ES ce AEEA SONER 7 Introduction ssessrossnssnsnnsnrsnsonsnrsnsnnsnnsnsnnsnnsnsnnsnrnsnnsnrsnsnnnssssnsnnsnsnssnssnsnssnssssnsnarsesnesarsesnenane 9 System OVEVI Wii DREES Gen ciao ee 11 Enhanced Security mode and 21 CFR Part Il 12 2I CEFER Part leia ta was 12 Enhanced Security mode nes et EEE SERES 12 Software installation eerste do 13 Operation Overview eva aaee E EE ans 14 Operating PerkinElmer 2030 c cossosssonsensssnsesnnessssssssonssonsnonsnnssnnsennssnssnnsennssnnnsnsnnnnesense 15 Starting up PerkinElmer 2020 17 Without enhanced security mode 17 With enhanced security mode 17 Live displaye EE 18 Kinetic Chart
34. 3 Press the button briefly 6 Briefly press the button on the reader this will be blinking if there are more plates to be loaded for this assay If it is not blinking then the assay has finished 27 Operating PerkinElmer 2030 Blinking light shows assay Continue until all plates have been measured 28 Operating PerkinElmer 2030 Starting operation with the Start Wizard In the enhanced security mode the Start Wizard is enabled only for users who belong to the Operator user group An alternative way to start counting is to use the Start Wizard This allows you to select any existing protocol to define how many plates you are going to use and to give the positions of samples Alternatively you can create a new protocol Proceed as follows Click on the Start Wizard icon and follow the instructions that appear PerkinElmef 2030 Manager Eile Instrumegt Tools Help The Start Wizard will appear to guide you through the procedure for starting The steps when using an existing protocol are Introduction to the wizard You can select the introduction not to appear next time if you want Protocol selection select the protocol you want to use for measurement by checking use existing protocol and selecting the protocol from the protocol tree 29 Operating PerkinElmer 2030 PerkinElmer 2030 Start Wizard Use existing protocol Create new protocol Lg Protocols Eg PerkinEl
35. 33645 313583 862650 159646 6820955 570487 549579 598887 589612 787267 552178 913831 807480 966402 678711 337029 735693 791044 274427 326696 210948 710979 309806 876276 830321 64595 408060 162063 775968 92542 163755 95473 580548 33354 932775 564474 634388 446552 198561 918121 434859 963683 975557 6826575 826877 774578 452897 489243 12507 266541 992356 483291 264124 582179 621457 413680 339778 103057 93411 113999 A7744 SERENA 277004 29054 17916 204 2223124 38763 ARAIN 264529 4 List Plates 1 1 A Plate Paget A Protocol A Errors A Notes Ready Errors may have occurred in measurement Check results Plate 1 of 1 Repeat 1 of 1 The cells in the two first rows displayed include information about the currently displayed plate The information included is Plate The number of the physical plate currently being displayed Repeat The number of the repeat of the physical plate currently being displayed Measurement end time Date and time when the measurement of the result plate was finished Start temperature Plate holder temperature when the measurement of the plate was started End temperature Plate holder temperature when the measurement of the plate ended Barcode Barcode of the plate if used if not used it displays the text N A 68 Result viewing The cells displayed in the rows directly above each resul
36. 50 151 153 Folders 44 G General options 128 G factor 90 H Heating 52 Height 51 Height of the plate Plate type 126 How to change filter slide settings 105 ID Protocol editor 53 ID tab 49 53 Injector position during measurement 57 Install of output option in MultiCalc 144 Installation of software 13 Instrument name 127 Instrument options 74 130 Instrument properties 127 K Keywords 59 Kinetic Chart 17 21 Kinetics button Measurement tab 57 L Label button Measurement tab 56 Label name 81 84 Labels 73 75 Labels selecting 54 Labels window buttons 75 LANCE 91 LANCE normalization 112 117 LANCE normalization wizard 73 Latest assay run 65 73 Latest assay result icon 19 Legend 20 Level user 74 Light exposure area LANCE 92 Light Exposure Ref Level LANCE 93 Light Exposure Reference Level TRF 79 Light exposure time TRF 78 Light on the button 40 Linear scale 21 List Result Viewer 67 Live display 18 19 27 Live Display 17 Logarithmic scale 21 Luminescence counting time 87 Luminometry 86 Measurement 54 Measurement Protocol editor 54 Measurement height Plate parameters 51 Measurement mode Plate 54 Single well 54 57 Strip 54 57 Measurement mode Measurement tab 54 Measurement operations list Meas tab 55 Measurement status page Eu Sm norm 110 Meas status page LANCE norm 114 Measurement tab 49 54
37. CE option installed Europium chelate emission LANCE option installed Samarium chelate emission Terbium chelate emission Dysprosium chelate emission Colorimetric label emission Colorimetric label emission Colorimetric label emission Fluorescein excitation Fluorescein emission Umbelliferone excitation Umbelliferone emission Name D340 D320 D615 D665 D642 D545 D572 P405 P450 P490 F485 F535 F355 F460 Centre Wavelength CWL 340 nm 320 nm 615 nm 665 nm 642 nm 545 nm 572 nm 405 nm 450 nm 490 nm 485 nm 535 nm 355 nm 460 nm Bandwidth BW Full Width Half Maximum ca 50 nm ca 80 nm ca 8 5 nm ca 7 5 nm ca 6 5 nm ca 7 5 nm ca 7 5 nm ca 10 nm ca 6 nm ca 6 nm ca 15 nm ca 25 nm ca 40 nm ca 25 nm Maximum transmittance at peak wavelength Tpk ca 60 ca 70 ca 80 ca 80 ca 80 ca 70 ca 70 ca 45 ca 30 ca 30 ca 60 ca 50 ca 35 ca 50 Note The time resolved filters are available only if this option is included with the instrument and Terbium and Dysprosium filters are not included in instrument by default 106 Tools menu EuSm Dual label normalization Note TR FIA must be installed for this to be enabled In the enhanced security mode this tool is enabled only for users who belong to the Editor user group Before dual label europium samarium time resolved fluorescence measurements can be made the instrument must be normalize
38. E time resolved measurements can be made the instrument must be normalized for the protocol to be used This procedure corrects for the following effects in LANCE measurements background from the plate crosstalk of the donor signal into the acceptor window and quenching The final result is normalized to the maximum donor intensity To begin this procedure select LANCE normalization from the PerkinElmer 2030 Manager Tools menu This will activate a wizard that will guide you through the normalization procedure First comes a welcome to the wizard page PerkinElmer 2030 TRF LANCE Normalization Wizard Welcome to the PerkinElmer 2030 TRF LANCE Normalization Wizard LANCE Normalization Wizard is a tool you can use to create a persistent normalization for a LANCE protocol Cancel Help You can select that this does not appear in the future Click Next to proceed After this the main steps in the normalization are 112 Tools menu Protocol selection You must select from the list of LANCE protocols the one you want to do the normalization for Highlight the one you want and then click the Next button PerkinElmer 2030 TRF LANCE Normalization Wizard d Select a protocol to be normalized Protocol DS LANCE 615 665 N A LANCE 6157665 with normalization N A N A LANCE Long Decay 615 665 N A LANCE 545 572 N A LANCE cAMP kit N A LANCE Tyrosine Kinase Start up Reagent Package N A LANCE
39. E wizard are stored 118 Tools menu Plate dimension wizard In the enhanced security mode this tool is enabled only for users who belong to the Editor user group A description is given here of how the Plate dimension wizard can be used to fine tune the plate dimensions This is needed in the case of user defined high density plates 384 or higher to ensure exact positioning of the plate The factory default plates cannot be changed If the same plate type is used for more than one technology it is recommended that you repeat this procedure for each technology In this case when defining plates give each plate type a different name for each technology First a welcome page appears PerkinElmer 2030 Plate Dimension Wizard Welcome to the PerkinElmer 2030 Plate Dimension Wizard This wizard helps you to adjust the dimensions of a plate type by scanning some predefined wells and showing the results graphically This allows you to fine tune the well positions in a plate When the plate dimensions are set correctly you get more accurate results from the instrument NOTE You can only modify the dimensions of a user defined plate type If you want to change a factory preset plate type make a copy of it and run the wizard again e _ Hee You can select that this page does not appear any other time by removing the check mark from the check box Then click the Next button 119 Tools menu
40. T 163 Contents Trademarks DELFIA is a registered trademark and LANCE and Wallac are trademarks of PerkinElmer Inc Windows and Excel are registered trademarks of Microsoft Corp in the U S and other countries Introduction 10 Introduction System overview Note This manual describes all the features of PerkinElmer 2030 Some versions of the instrument do not have one or more of these features You can ignore the parts of this manual not relevant to the version of PerkinElmer 2030 you have PerkinElmer 2030 multilabel reader is a complete platform for quantitative detection of light emitting or light absorbing markers Depending on the model of the instrument you have it is suitable for flash or glow luminometry fluorometry fluorescence polarization high sensitivity time resolved fluorometry homogeneous time resolved fluorometry LANCE option and photometry It is a compact bench top unit with features such as dispensers shaking temperature control top or bottom reading and scanning The software is a 32 bit application running under Windows XP Up to 1536 well plates can be counted depending on the technology and model as well as Petri dishes slide filters and Terasaki plates Plates can be loaded manually or with stackers for automatic operation Output can be to a file on the PC and or to a laser printer 11 Introduction Enhanced Security mode and 21 CFR Part 11 Note Use of the Enhanced Securi
41. T ES o Eees SE 112 Using the LANCE normalization Wizard nen 112 Protocol selection 2 3 22 e E E E E E E E ea 113 Preparingithe pl te se ir 113 Measurement status page 114 Measurement warning Page unseeseesensesnensennensesnnnsonnnenennsnonsensennonsensnnonnenennsnonon 115 Normalization confirmation page occcoccconoccnonocnnoncnncononononcnnnonnon non cn nonnnn nero conc onnrnncn aran 116 Using LANCE normalization samples in the assay sccessseeceeeeceseeeeeeceeeeenees 117 Plate dimension wizard o e o IL 119 Plate s lection iii iba it 120 Tech ology sel cti unse Sieg est eene 120 Preparins thepl te u a hrenet alas dee Eed d en 121 Define the centre point of the corner wells 122 Plate types and Well name Sie isa earne ane re e CEEE EE AE EEEE Sa 124 Plate ty pe nianse A 125 Plate naM EE 126 Number Of well TO WS iii in o Rp Lands 126 Number of well Columns sseni a 126 Height of the plate o 126 Strip orientation sip eiet rea ae aa Eeer ra sea EAE E EEE EEEE EA 127 Plate dimensions omnia a a aE E EES 127 RTE 127 KEE 127 Instrument properties eneret ea EE AAE E eE EERE ERENS 127 OPIO a a a a a EES 127 General curia A ics ats EE EE ies Bobs Eed 128 Instrument hard wate soie sarosa ee E nie ie ds 130 Measurement technologies 2 2 2 2282er gb 131 Contents A eene Eed ROA 133 File ment 2222 TON 135 Automatic logout een ed 135 Troubleshooting sscsecossossiscccocesseasocsssessoovosecse
42. Ultra KinaSelect Ser Thr kit N A lt Back Cancel Help Preparing the plate The wizard explains what samples you should put in the plate and where you should put them The strip to be used must be horizontal e g Al to A12 etc The plate used is the one defined in the protocol you previously selected Blanks should be in Al and A2 these are used to correct for the plate background in the acceptor window These are made by dispensing buffer only Spectral crosstalk samples 1 e donor only samples in A3 and A4 these are used to calculate the amount of the blank corrected donor signal in the acceptor window as a proportion of the donor signal Only donor and buffer should be dispensed to these wells High intensity samples in AS and A6 these are to normalize the final result to the maximum donor value The ratio of the high intensity donor value to the sample donor value corrects for possible quenching in the sample These samples should include all the reagents to give the maximum acceptor signal All other wells should be left empty Then load the plate and click Next 113 Tools menu PerkinElmer 2030 TRF LANCE Normalization Wizard Prepare the Generic 8x12 size plate plate so that 1 blank series is placed in wells 41 and A2 2 cross talk series is placed on wells A3 and A4 ER high series is placed in wells A5 and A6 and all others wells are left empty Load the plate into the instr
43. Userid Password combination with five user levels e improved file security covering protocols versioning and result files e audit trails of user actions This manual and the electronic help will tell you how to use PerkinElmer 2030 with or without the Enhanced Security mode All differences in use are noted and explained Please refer to Appendix 4 for details of special functions that are part of the Enhanced Security mode 12 Introduction Software installation Note The following instructions assume that the reader PC and printer have been connected up and switched on see the installation instructions in the Instrument manual If Enhanced Security mode is used then installation can only be done by a person with System and Windows administrator rights See Appendix 4 for more details 1 Load the software CD 2 The setup loader will start automatically and give you a choice of either selecting the Installation Wizard to guide you through the installation or reading files from the CD 3 When you have completed the installation you should remove the CD from the PC otherwise the setup loader will start again next time you boot the PC Note In Windows XP the user has to have either administrator or power user rights 13 Introduction Operation overview Parameters must first be set using the protocol editor This is activated by selecting a protocol in the Explorer or starting an assay with Start Wizard
44. ad a plate the button will be blinking See the picture on page 20 for correct plate orientation Load first plate Light is blinking 2 Press the button briefly Operation will start The button light will be lit without blinking The barcode on the plate will be read and the protocol specified by the barcode will be selected and used for measuring the plate When the measurement has finished and the light starts blinking again remove the plate and load the next one and briefly click the button If the plate belongs to the same assay as the previous one no barcode is needed If it is barcoded for a new assay then the appropriate protocol will be used for measuring the plate 34 Operating PerkinElmer 2030 1 Rernove the eg measured plate 2 Load the next plate 3 Press the button bnefly Continue measuring plates this way Note that the button light will remain blinking as long as the protocol number barcode option is selected 35 Operating PerkinElmer 2030 Stacker operation A stacker is able to hold up to 20 or 40 in the larger version microplates No lids loose frames or very loose strips are acceptable See Appendix 2 for a list of plates that fit in the stackers Starting operation no protocol number barcode model the loading area is uncovered when the stackers are not in place so you must avoid the danger of getting your fingers trapped by the plate lift mechanism If you need to do some
45. aiable yet For Help press Fi Temp C 25 7 Type the appropriate userid password combination to proceed Clicking the Cancel button closes PerkinElmer 2030 Clicking the Change password opens the Change Password dialogue where you can define a new password After a successful login PerkinElmer 2030 can be used according to the permissions granted to the logged on user Live display The Live display shows actual assay results measurement values on the wells if number of wells in the plate is 96 or less For all plate sizes if you move the cursor on to a well that has been measured the numerical value will appear At the bottom of the window there is information about the sample being measured including its current value Note In multiple label scanning and kinetics measurements only results from the first measurement of the 18 Operating PerkinElmer 2030 label are visible in the Live display You can see results from all labels in the Latest results display which you can get by clicking the Latest assay result icon The Live Display tab enables you also to control the instrument by starting stopping or ending counting It shows you the protocol being used to control the measurement and it is also possible to see the contents of any of the protocols by selecting the required protocol and clicking the View Selected Protocol button In the enhanced security mode these controls are enabled only for users who belong to
46. an choose where they are located on the plate as the following example shows 117 Tools menu E Copy of LANCE 615 665 with normalization Protocol editor Eile Help O ID Samples Measurement Outputs and Events Number of plates and mapping of each plate M4 pj aa Plate 1 of 1 re a E P E S Alf o m o 9000000 o 900000000 90000000 900000000 90000000 9000000 90000000 90000000 90000000 O 90000000 90000000 900000000 Plate parameters Measure each plate 1 1 300 times Plate type Delay between repeats 0 0 3600 sec Generic 8x12 size plate e r Measurement height Standard approx 8 mm User defined mm from bottom 13 0 18 0 mr r Temperature level checking JT Wan if plate holder temperature differs from 25 C Ready When you have defined the plate and saved it you can run the assay in the normal way When the LANCE samples are detected on the plate they will automatically be used to make the necessary corrections for this particular LANCE measurement They will overrule any values that may have be stored in the instrument memory from a normalization with the LANCE wizard However these on the plate normalization values will not be stored and used for any other assays than the current one Only values resulting from use of the LANC
47. are dra 57 Scanning e 57 Injector position during measurement uuncessessensesnersessensennnenenesnensennnsenne nennen 57 Outputs and Events u ii 58 LEET 59 File EE 61 EE 61 Print E 61 Print Oe 61 eko e EE 61 e EE 61 BK E UE 61 Result vieWingi scscssisssischsssssesussensannssnssusessusenennsninnens esnsensensnnsssheseresse nenn toonset aeaoe 63 Latest results ae air Re EE 65 Results in the Explorer soraio kann nia ee Ee EE e Ee 65 E EE 67 Plat map Aare elektr Silo 68 PrOLOCOM 22 2 eben esse pen en 69 le 69 EOL a Seen es 70 PO id 70 Export reli ii hr en 70 Contents Tools menu 2ussesneonenesnennenesnennenesnennenennennenennennenennennenennennenennennesesnennesesnennesesnennssesnesesnennenane 71 Ke OT UE 73 Userlevel ci ia ai SS 74 Labels Measurement Technologies 75 Time resolved fluorometry ccccceessessceesceescesecesecsecssecesecaeecaeeeaeeeneeseeeeseeesenseeseenaes 77 Label name cercos o EE Ee 77 Flash Energy EE 77 Flash Energy Leye 29 ease Eege Ee 78 EE Le 78 Lisht Exposure Linea Era nee eEen 78 ele 78 Light Exposure Reference Level 79 Emission filters Laia id 79 Emission apertures 335 253 aie esata aversion oe netting bean a E ETE ara 79 Counting parameter 79 Factory set values for the main counting Window nen 80 Be in Size iii ee EE Ei EIER ida 80 Second measurement parameters oconoccnocconnconnnonconoconoconocnnonononnnonnnrnn cnn oran nnnncnnccn recono 80 O 81 Babel nam
48. assword Control This tab allows the PerkinElmer 2030 administrator to set the following password related settings User Management Users Password Control Login History Audit Trail Summary Login Type PerkinElmer Login Bi Account Lockout Maximum password age c 2 lt Minimum password age Allow changes immediately e Allow changes after days 1 3 Minimum password length At least characters 6 Password uniqueness Number of passwords to remember 24 3 2 OK Cancel Maximum password age defines the maximum number of days that users can retain the same password before they must change it By default the password expires after 42 days The minimum is 1 day and the maximum is 999 days The Maximum password age cannot be set less than or equal to the Minimum password age If it is not necessary for users to change their password select Password never expires Minimum password age defines the number of days that users must retain the same password before being allowed to change it The default is to Allow changes after 1 day 157 Appendix 4 Enhanced security special functions This prevents users from changing their password several times in a short space of time in order to return to a previous password To allow users to be able to change their password immediately select Allow changes immediately The Minimum password age cannot be set
49. asurement tab 55 Dual label 48 Dual label normalization 107 Dual label normalization wizard 107 Index E Editor 154 Eject slide button 101 Emission aperture 89 Emission filter slide 100 Emission filters 79 82 85 86 89 93 97 Empty 150 Enhanced security 12 Archive 159 Audit Trail 157 Password control 154 155 156 Permissions 154 Special functions 154 User groups 154 User management 154 155 Error Result Viewer 70 Eu Sm dual label selection 48 EuSm Dual label normalization 73 107 EuSm Dual label normalization wizard 107 Excitation aperture 82 85 Excitation filter LANCE 92 Exit 135 Exit Protocol editor 61 Explorer 43 73 Explorer folders 44 Explorer icons 47 Export file per plate 129 Export results Result Viewer 70 F File export 70 File menu 135 Automatic logout 135 File menu Protocol editor 61 File types for export 70 Fill 150 151 Filter CW Lamp 82 85 89 98 Filter emission 79 82 85 86 89 93 97 Filter excitation LANCE 92 Filter properties 106 Filter slide settings changing 105 164 Filter slides 99 Filter wheel CW Lamp changing 104 Filters 73 96 Filters window buttons 96 Fine tuning plate dimensions 119 Flash Energy Level TRF 78 92 Flash Energy Mode TRE 77 92 Fluorescence counting time FP 89 Fluorescence counting time Fluorometry 83 Fluorescence polarization 88 Fluorometry 81 Fluorometry tab 81 Flush 1
50. atus of the measurement The progress of the measurement is shown by the progress bar The Next button becomes available when the measurement has been completed When the measurement is completed click Next to move the wizard directly to the Normalization confirmation page otherwise the wizard moves to the Measurement warning page Measurement warning page This is used to inform you that something out of the ordinary has happened during the measurement of the normalization assay This page gives you enough information to decide whether the results can be accepted regardless of the problem that caused the warnings Clicking Next on this page advances the wizard to the normalization confirmation page Clicking Cancel closes the wizard and the normalization factors are not recorded in the database 110 Tools menu Normalization confirmation page o PerkinElmer 2030 EuSm Dual label normalization Wizard The normalization measurement is now completed Check the results and press Finish to accept the new normalization factors or Cancel to ignore them Normalization factors Ratio 1 4433 Blank 396904 Supplementary values Sample Label Mean CV Blank Samarium 492662 55 1 Blank Samarium in Europium window 400157 65 5 Europium Samarium 387696 70 6 Europium Samarium in Europium window 472885 56 1 Help This appears after normalization has been successfully completed It shows the mea
51. ay light getting to the photomultiplier tube Counting parameters These determine the time factors involved in the measurement process all times are in microseconds Counting delay the time after the excitation pulse at which counting of the emission signal begins Counting window the time period within which counting occurs Counting cycle the time between excitation pulses within a measurement The second or additional counting window is fully independent of the first one and it useful when optimizing time resolved parameters or for calculations of the decay time of the label 79 Tools menu Factory set values for the main counting window Label Delay Window Cycle Eu 400 400 1000 Sm 50 100 1000 Tb 500 1400 2000 Dy 30 30 1000 Factory set values for the second counting window are always 0 0 The counts in this window are displayed only if the values deviate from zero Beam size This parameter can be selected to be Normal or Narrow provided the Adjustable beam size option has been installed It enables measurements on plates with very small well sizes by focusing the excitation light beam to a smaller size Second measurement parameters In the case where you want to make measurements with two filters you can use this parameter to select the type of the second emission filter The options are the same as for the first measurement emission filter Two measurements are made first with the normal filter and then with the se
52. button will be activated Click it to continue Define the centre point of the corner wells PerkinElmer 2030 Plate Dimension Wizard Left click the mouse at the center of each well and then click Next Weit well2 wera wei 1 2 3 4 5 6 7 3 9 203226 228622 177829 203226 E 152433 177829 127036 152433 01540 127036 76243 101640 50847 76243 25450 50847 54 25450 VARIA ANO coca zm The window that appears allows you to define the centre point of each of the four corner wells 122 Tools menu Each tab shows one of the four wells with a plot of the intensity of the light from the well This is colour coded with red showing the highest intensity Position the mouse cursor where you think the centre of the well is and click the left mouse button A black cross will appeared centred at the point of the cursor If you are not satisfied with the position of the cross move the mouse cursor and click the left button again Repeat this for each of the four wells When the centre of all four wells has been marked the Next button will be enabled Click this to continue PerkinElmer 2030 Plate Dimension Wizard Check the errors of the well center points and modify them if necessary Click Finish to save the new dimensions or Cancel to ignore them Deviation of well 1 Deviation of well 2 Horizontal 0 513 mm Horizontal
53. cond which the system automatically moves into place 80 Tools menu Fluorometry Fluorometry light of a particular wavelength is selected with a filter from the spectrum of a continuous wave lamp and used to excite the fluorochrome in the sample The parameters that can be set for fluorometry are the following Fluorometry Label Properties Name Fluorescein D Del Password Last Edited No user 2 15 2008 1 38 43 PM Cw Lamp Energy 5028 BE RR tear Cesta CW Lamp Control Stabilized Energy Constant Voltage DA Lamp Filter F485 Slot A5 v Excitation Aperture C Small Normal C Large Emission Filter F535 Slot A5 r Emission Aperture C Normal Small Large Counter position Top Bottom Counting Time 0 1 600 5 Second Measurement CW Lamp Energy DA Lamp Filter d y Emission Filter y Cancel Help Label name This is the name by which the label is identified A password can be associated with it to prevent parameters being changed by an unauthorized person CW Lamp energy Using this parameter you are able to change the excitation energy supplied by the CW Lamp 81 Tools menu CW Lamp Control Mode There are two ways of ensuring that the light incident on the plate from the CW Lamp remains constant stabilized energy mode or constant voltage mode In the stabilized energy mode a photodiode close to the light path has part of the incident light dire
54. cted onto it This sends feedback to the lamp controller which alters the lamp voltage to adjust the intensity of the light produced so that it remains constant In the constant voltage mode the lamp voltage is not changed but the software corrects for changes in the lamp output You can select the control mode you want from the option buttons The default mode is stabilized energy The use of the constant voltage mode allows the CW Lamp lamp to be run continuously at maximum power and it is faster than the stabilized energy mode This enables optimum performance in cases where the light intensity is critical as in fluorescence polarization or where speed is critical as in fast kinetics CW Lamp filters You can select the CW lamp excitation filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory pre set CW lamp filter names begin with a letter indicating the technology for which they are to be used F is for fluorometry After the letter comes the wavelength Excitation aperture PerkinElmer 2030 0050 only There are three sizes of excitation aperture controlling the size of the incident light beam You can select with the option buttons Small Normal or Large Normal is the default Emission filters You can select the emission filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factor
55. d Protocol Editor In the protocol editor dialogue select the ID tab Select the protocol number to be the same as the MultiCalc protocol ID number Then under the Outputs tab include all items in output and select the File type as MultiCalc The File Name should be as follows c wiacalc Ocom 1420_2 lt ProtocolNumber gt lt RunIDpad2 gt Note Max 3 labels can be selected Plate repeat must be 1 Kinetics or scanning measurements are not possible Plates having more than 144 wells are treated sequentially in MultiCalc 143 Appendix 1 Connecting MultiCalc to PerkinElmer 2030 Installation of PerkinElmer 2030 output option in MultiCalc Copy the files 1420_2 C02 and 1420_2 h2 from the distribution CD ROM and paste them into c wiacalc Ocom Set the properties for 1420_2 in MultiCalc to AOSPMTV by selecting on level 5 ETC SYSTEM ETC COMM PROT PROPERTY select 1420_2 type the letters AOSPMTV and press Enter Escape to the main menu and then restart MultiCalc Note Property A stands for Automatic evaluation whenever an assay file is available Property O is for the connection type Offline MultiCalc assay protocols All assays with PerkinElmer 2030 have to be of the FLUORO type for all measurement technologies used in PerkinElmer 2030 The response from labels 1 and 2 will appear as COUNTS and COUNTSB and from label 3 as COUNT Usage of MultiCalc Leave MultiCalc in the counter mode evaluation will occur automa
56. d allows you to specify on what unit the complete measurement sequence should be performed before moving on to the next similar unit The possible unit is a single well a strip or a plate Plate and strips modes are disabled when using a fast kinetic or scan measurement in the operation sequence Well mode will be disabled when using more than one label in the 54 Protocol editing operation sequence However copying and pasting of the same label is possible Plate mode can be used for slow kinetics by making repeated counting of the plate with delays in between Measurement operations This field lists the operations that have been specified to be used in the assay The operations are listed in the order they will be performed If you select an item in the list and then click the right mouse button a menu will appear to allow you to Copy Delete or see the Properties of the selected item A row of buttons beside the Operations list enables you to add items to the list Click the button referring to the operation you want Set the parameters and click OK to add the item to the list Down button This moves the selected operation in the operation list one step closer to the end This button is enabled when the operation currently selected in the measurement operations list is not the last operation in the list Up button This moves the selected operation in the operation list one step closer to the beginning This button is enabled when
57. d for the plate type s to be used To begin this procedure select EuSm Dual label normalization from the PerkinElmer 2030 Manager Tools menu This will activate a wizard that will guide you through the normalization procedure First comes a welcome to the wizard page O PerkinElmer 2030 EuSm Dual label normalization Wizard Welcome to PerkinElmer 2030 EuSm Dual label normalization Wizard Normalization Wizard is a tool you can use to quickly normalize a plate to be used in EuSm Dual labelmeasurements Cancel Help You can select that this does not appear in the future After this the main steps in the normalization are Plate type selection You must select from the list the plate type you are going to use This is necessary because normalization is plate specific i e you must have the same size and type of plate as you plan to use for the actual dual label measurement The list shows Plate type name Date when the normalization for this plate type was last done Name of the user who ran the last normalization 107 Tools menu Correction and background values from the last normalization run O PerkinElmer 2030 EuSm Dual label normalization Wizard Select plate type to be normalized Plate name Generic 8x12 size plate Labsystems 8x12 DELFIA strip plate NUNC 8x12 DELFIA strip plate Costar 3512 3x4 wells Costar 3524 4x6 wells Falcon 3047 4x6 wells NUNC 384
58. d in PerkinElmer 2030 because you may jam the conveyor If the button on PerkinElmer 2030 is blinking it means that the assay has not finished and you must remove the empty input stacker load an empty output stacker and then an input stacker which has the remaining plates of the assay plus plates for other assays if you want 37 Operating PerkinElmer 2030 Starting operation and using a protocol number barcode Caution Do not put your fingers into the sample loading area In the stacker model the loading area is uncovered when the stackers are not in place so you must avoid the danger of getting your fingers trapped by the plate lift mechanism If you need to do something in that area switch power off first This configuration makes best use of the stackers operation is automatic 1 Load the stackers The button light will be blinking Light is blinking 2 Click the button briefly One by one all the plates will be loaded and measured If a plate has a barcode then it will be measured using the protocol specified by the barcode If it has no barcode it will be measured using the protocol specified by the previously read barcode 3 Only when the input stacker is empty will operation stop The button light will be blinking Remove the stackers and load new ones if required 38 Operating PerkinElmer 2030 4 Click the button briefly to start operation again Ending operation Stop button Clicking the Stop button i
59. d including a number of default types Well names the available names are listed There are up to five default types two Empty and to be Measured that are generally valid and three LanceBlank LanceCrosstalk and LanceHigh for on plate LANCE normalization measurements these require that LANCE is installed Note Well names cannot be copied Instruments when you select this tab you can see names of instruments Selecting a name and properties gives more information The following buttons appear in Plate types and Well names 124 Tools menu Add add a new item to the list available e g a new plate type Give the name and edit the properties It will appear with a new icon showing it is different from a default item Copy select an item to activate this Make a copy of an item It will appear with a new icon showing it is different from a default item and the name will be preceded by the words Copy of You can give it a different name and edit other parameters by selecting Properties Remove select a user created item to activate this Remove the selected item from the list You cannot remove a preset item from the list nor an item that is used in a protocol This command requires confirmation because you cannot undo Remove Properties select an item to activate this View the properties of the selected item If the item is user defined then you can edit the properties but if it is a factory pre set you cannot e
60. dialogue will appear New User User User name Eull name Password Confirm password Status Enabled Disabled All the fields in the New User dialogue must be filled in Status enabled disabled sets the login status of the new user Status disabled means that the user cannot log in to PerkinElmer 2030 Manager Status enabled default means that the user can normally log in to PerkinElmer 2030 Manager The User must change password at next login checkbox is checked and disabled This means that the Change Password dialogue appears after the next successful login and the user has to change the password Select the user from the drop down list and click Edit to change the settings of the selected user e g password The Edit user dialogue will appear The functionality is similar to that of the New User dialogue except that the user name cannot be changed and the User must 156 Appendix 4 Enhanced security special functions change password at next login checkbox is enabled and not checked When you are enabling a disabled user you must also enter a new password for that user Select the user from the drop down list and click Delete to remove the selected user Confirmation is asked Select the user from the drop down list and define the group membership for the selected user by using the Add and Remove buttons A user can belong to several groups P
61. dit them Plate type When you click the Plate types tab the available plate types are listed This list includes a number of default types To define a new plate click Add You can also click on an existing plate type and then click Copy to save the plate type with a new name If you wish you can edit the parameters to make a new plate type Click Properties to see the plate properties 125 Tools menu Plate Properties Name DptiPlate 96 Parameters Number of well rows le 1 32 Number of well columns h2 1 48 Height of the plate mm has Strip orientation Ze Horizontal Vertical Dimensions from the corner of the plate to the center of the first well and from the center of a well to the center of the next well mm Tune OK Cancel Help In the case of Properties the dialogue that appears gives information about the plate Each of the parameters in this dialogue must be defined when you add a plate The software does not allow you to give unreasonable values e g a well separation that would mean that all wells do not fit in the plate The maximum number of rows and columns are shown Plate name This is the name that will appear on the list of plate types You can change this name by editing the current name or by marking it and deleting it and typing a new name Number of well rows This is the number of wells in a row The product of this with the number of well columns giv
62. e seconds have elapsed since the page was opened During the time the Finish button is disabled the text of the button is Wait t where t is the remaining waiting time and it is updated every second When you click Finish the new values replace the old values in the database 116 Tools menu Using LANCE normalization samples in the assay Instead of running the LANCE normalization wizard and saving values that can be used for any subsequent LANCE assay with that particular protocol you can include LANCE normalization samples on the plate as part of the assay samples and thus obtain corrected values for that particular plate When you define the protocol make sure that both LANCE labels are defined in the Measurement tab as shown below If only one label is selected then the measurement will be treated as normal time resolved fluorometry I Copy of LANCE 615 665 with normalization Protocol editor LK File Help ID Samples Measurement Outputs and Events Measurement mode By plate C By wells C By strips Operations Operation properties unce 615 Lauren eelalalsix a Ready When you set up the plate you can select the LANCE well types from the menu that appears when you click the left mouse button Use this feature to include at least one of each of the three LANCE well types on the plate You can choose the number of replicates of each well type and you c
63. e eg oi EES ee Ee 81 CW Lamp energy unse ERENNERT EEN 81 CWw Lamp Control Modest ansehen 82 CW Lamipifiltels cia 82 Excitation aperture PerkinElmer 2030 0050 only 82 Emission filters an rear Qaida Bia ei been 82 Emission apertures ss wis ists dute sees Beate E Leelee aes 83 Counter DODERER 83 SR SEENEN ERC DEES E EEEE SEAE E VESETAS 83 Second measurement parameter oconoccnonconnconcnonconnconocnnocnnonononononnnrnn cnn conc nnnncnnccn recono 83 Ph tomeltry cuisine lena E cede eiii Bruns deca EE 84 Label Names ica dress 84 Absorbance mode scoet roe ita iia 84 Flash Lamp O SE 84 CW Lamp filters 222 a A 85 Excitation aperture PerkinElmer 2030 0050 only 85 REA time Al a bares ede 85 Second Measurement rain rasieren 85 Eummometry a nlaensbeia rs ih insbes 86 Label name 0 ia 86 Emission filter 86 Contents Emissi n apertute an anche cast Settle REENEN 86 SON 87 Second Measurement errors 22 4 better Es ee dan dencia OE liada ds 87 Fluorescence polarization PerkinElmer 2030 0050 only 88 Label mamen Ee eene Ee ENEE 88 CW Lamp energy nosar ene iria ri ii 88 AA ssets cscs ecsuepco cesses catbintie 2h ss sesesedonststs ssbansaves iisthasssenayionidvvashasssetts 89 Pol rizer pert rer 2 2 aus haa Gates its Saves 89 Emission filter ihe ea Reigate hes eee 89 Emission E 89 COLUMNS Meca a 89 DIE adi iia aa 90 LANCE canal a 91 Labelname it a 91 Flash Enerey Modesto toda rte Seed Gg 91 Flash Energ
64. e measurement is completed These are the same as for printer output List Plate Protocol description Error description Notes You must also type in the file name This can include keywords connected with the file as described in Events The button to the right of the filename box gives you a list of all valid placeholder arguments to choose from Other filenames possibly used in other protocols are also available through this button Note You will not be allowed to change to another tab if an invalid placeholder argument is found However if a previous file with the same file name exists it will be overwritten by the new file without warning Under this tab you can give a command that will be executed when a protocol has been run The keyword should be in triangular brackets lt gt exactly as the examples given in the tab show These commands are then automatically replaced by the appropriate name or number etc after the assay run ends The first box specifies the executable to run while the second box specifies the command line arguments to use The button to the right of the first box lets you browse to find the executable file you want to use The button to the right of the second box again lists all the valid keyword arguments that you can choose from see Keywords below For example you can start an external spreadsheet running and have it execute the file produced by the run This file will be identified by e g protocol name
65. e output File output options File type Excel 5 0 x Filename ml Ed Include in List output Plate Protocol description Eug description Notes Assay end system command This command will be performed when the instrument has finished measuring the assay using the edited protocol Ready The Outputs tab in the Protocol editor allows you to select where results will be sent at the end of arun This can be to a printer and or file Click the check box of the output device s you want If you have selected Printer output you must select the format either plate format results are in a table format with rows and columns in the order shown in the plate map and or list format results are in a single column starting with those from the first well When this option is not set the Include in output checked list box is disabled This list box defines which parts of the assay run data should be included in the printed output List Plate Protocol description Error description Notes If you have selected File output you must specify the file type from the drop down menu of possible types The types you can save files in are Excel 5 or later versions Tab delimited text or MultiCalc 58 Protocol editing If the Include in output list box is checked then it specifies which optional parts of the assay run information should be included in the file saved when th
66. e output and give a command to be followed after an assay has been run with this protocol You can set parameters for up to 40 plates in one protocol Protocol entries are checked for validity by the software and invalid entries are informed when you try to switch from one view to another When you have edited a protocol you can use the command line or the button bar to save or print the protocol Note If you want to change the plate type do this before setting other plate parameters This is because when the plate type is changed all the previous plates will be deleted and one plate of the selected type will appear 49 Protocol editing Samples B kinetic Protocol editor BAF Samples Measurement Outeurs and Everts Number of plates and mapprag of each plates mln ala Piste 1 of 1 tf 2 3 4 s 6 7 e 9 0 1 12 eeeeeee000000 000000000000 000800088888 LEI 0000008890808 000 00080008 000 900000000000 Plate purametess Vespa each plste T 1 20 times Plate yox Delay between geg 0 0 2600100 Generic bi size pite sl Maspaeert Iert Standard soeren Omm C Urs drei wm from beiten Temper level checking TF Wam plate holder temperahune diftess hom Selecting this tab causes a plate map to be displayed to allow you to edit samples Buttons allow you to select the first plate previous plate next plate or last plate You can also add a ne
67. eated by WorkOut data analysis software if that is installed on the system To open a folder click on it and then select Open from the File menu Alternatively you can open a folder by double clicking the left mouse button when the cursor is on the folder icon You can also click once on the mark next to the folder to see the protocols contained in it To close a folder either double click on it or click the sign next to it If you are working on the Advanced or Service level you can create rename or delete folders A sub folder of Users can be renamed by selecting it and clicking the right mouse button and selecting Rename Explorer views Different view icons are provided to display data related to the currently highlighted protocol or the assay result x 9 8 8 Bam m This icon is for Original View In this view protocol tree is shown in the left pane and the assay results are shown on the right pane 44 Protocol editing gx aa gt e Slam e Tegn data Elie D Tror Wm 12 00 07 AMAZON 1112 17 EM DENO DATA GENERATED 348 2008 1 26 33 94 DENO DATA GENERATED This icon is for Protocol View In this view details of the highlighted protocol are displayed If an assay result is highlighted the view displays protocol and assay result in a single active window 45 Protocol editing yasuring Sequence ame Zabel Absorbance Protocol created by sE Protocol creation date
68. ecting the features accessible to the user Users appears only in the enhanced security mode for user management functions Audit Trail appears only in the enhanced security mode Archive appears only in the enhanced security mode Options includes demo mode selection barcode and plate map features 73 Tools menu User level Note In the enhanced security mode extra menu items appear in the Tools menu The User level this does not appear in the enhanced security mode in the Tools menu allows you to select the user level Routine Advanced or Service Tools Help User Level f Explorer User level in the PerkinElmer 2030 system defines what Start Wizard options and commands are visible in the Workstation J software user interface Possible choices are Latest Protocol run Latest Assay Result r User level Routine Only features needed for day to day J View Picklist Protocol basic measurement usage are enabled a bel Advanced Allows editing of Settings filters etc Labels From time to time there may be a need to Filters configure the system because the 4 requirements for day to day usage have Plates and Wells Settings changed j This user level also enables some i gt ati advanced editing commands in Explorer EuSm Dual Label Normalization eg protocol group dag amp dopl LANCE normalization isa H BM For authorized service personnel only Plate dimens
69. ector to let you specify a label to be inserted in the measurement operations list There can be up to 20 labels in the list Note For LANCE measurements two labels must be selected If only one is selected then the measurement will be treated as normal time resolved fluorescence 56 Protocol editing When using well or strips mode the label button will be disabled when there is already a label in the measurement sequence However copying and pasting of the same label is possible For detailed information about labels see Labels on page 67 Kinetics button Clicking this causes a dialogue to appear that lets you specify the parameters for a kinetic measurement You can give the number of repeat measurements up to 300 and the delay between each measurement 0 to 600 s You need to also select the label by clicking the label button and then selecting from the label dialogue If the repeat is less than 26 and the delay between repeats is 0 results will come in one group The kinetics button is enabled only when using well mode and there are no other labels in the measurement sequence However copying and pasting of the same kinetic label is possible Scanning button Clicking this causes a dialogue to appear that lets you specify the parameters for a scan measurement This means that several measurements are made for a single well and the measurements are made from different points of the well These points are in the form of an array for
70. efined otherwise it is not available N A and also any notes that have been included in the protocol When you create a protocol you can save it as a user protocol in the Users folder You can move a user made protocol to a different folder by selecting it holding down the mouse button dragging it to the target folder and releasing the mouse button Note The dual label protocol only available if TR FIA is installed should not be copied and edited e g to make a single label europium or samarium protocol You should begin with an existing single label protocol For dual label operation you must select the europium label first and then the samarium label After you have selected the second label a third samarium in the europium window will be created automatically When running with these three labels you must have a valid Eu Sm normalization see the Tools menu for how to make such a normalization For information about results see the chapter on Result viewing 48 Protocol editing Protocol editor Note Changes to protocols should only be made by authorized persons The protocol editor window contains four tabs ID you can identify the protocol and you can see information about the protocol Samples shows a plate map to allow you to edit wells and plates and select the plate type and other parameters Measurement you can select the labels and other operations used for measurement Outputs and Events you can define th
71. el Help Label name This is the name by which the label is identified A password can be associated with it to prevent parameters being changed by an unauthorized person Emission filters You can select the emission filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory preset emission filter names begin with a letter indicating the technology for which they are to be used L is for luminescence After the letter comes the wavelength Emission aperture In front of the photomultiplier tube there is a 4 position aperture slide which has three different apertures and a shutter position 86 Tools menu The normal aperture is circular with a diameter of about 4 mm and it is used for factory set labels The small aperture is circular but only 1 mm in diameter This allows you to shift the dynamic range of the emission signal for a particular user selected label which may otherwise exceed the linear range of the instrument This aperture is used for the top fluorometry factory set labels There is also a large circular aperture 5 mm in diameter The shutter is used at all times when no measurement is occurring to prevent stray light getting to the photomultiplier tube Counting time This parameter determines how long the luminescence is counted for Second Measurement In the case where you want to make measurements with two filters you ca
72. ement The second or additional counting window is fully independent of the first one and it useful when optimizing time resolved parameters or for calculations of the decay time of the label 93 Tools menu Factory set values for the main counting window Label Delay 1 Delay 2 Window 1 Window 2 Cycle High count 615 50 0 100 0 1000 High count 665 50 0 100 0 1000 Long decay 615 50 0 100 0 3000 Long decay 665 50 0 100 0 3000 545 50 0 100 0 3000 615 50 0 100 0 1000 665 50 0 100 0 1000 572 50 0 100 0 3000 TruPoint 615 50 200 100 100 1000 TruPoint 545 50 200 100 100 1000 Beam size This parameter can be selected to be Normal or Narrow provided the Adjustable beam size option has been installed It enables measurements on plates with very small well sizes by focusing the excitation light beam to a smaller size Second measurement parameters In the case where you want to make measurements with two filters you can use this parameter to select the type of the second emission filter The options are the same as for the first measurement emission filter Two measurements are made first with the normal filter and then with the second which the system automatically moves into place LANCE Label Properties e LANCE 615 100 flashes window 50 100 1000 615nm This label is for fast decay Eu chelates for example Eu W1024 in a TR FRET assays e LANCE 665 100 flashes window 50 100 1
73. empty and if not to empty it proceed as follows Open the slider on the front of the dispenser Remove the waste vial Empty out any liquid following the disposal procedure recommended by the manufacturer of the reagent you have been using Then replace the waste vial on its holder and close the slider This is important because it prevents light leakage to PerkinElmer 2030 148 Appendix 3 Dispenser operation Preparing the tubing Put the tubing into a container of water 149 Appendix 3 Dispenser operation Dispenser maintenance Note In the enhanced security mode this tool is enabled only for users who belong to the Operator user group PerkinElmer 2030 Manager e File Instrument Tools Help le ele Clicking the Dispenser maintenance icon or the item in the Tools menu invokes the Dispenser Maintenance Setup page to perform common dispenser maintenance operations This is enabled only when working with a system that has the dispenser installed The current state of the instrument server is shown at the top of the page y Dispense Maintenance E Instrument status dle Select pumpls and operation C Fill Empty Flush 500p 500 500pl 500 ph u m L The setup page presents you with a graphical representation of the number of pumps 1 4 available when performing an operation You can select the pump s which are going to perform the operation by checking b
74. ept an assay ID as large as 1234 If lt AssayID gt 234 and you add pad 4 to lt AssayIDpad4 gt the result would be 0234 This would be useful if you needed a four digits identifier The validity of possible keyword arguments is checked when you try to switch from this view to another If an invalid argument is found you are notified and the page switch is cancelled 60 Protocol editing File menu The protocol editor file menu contains the following items of which Save and Print have corresponding buttons Save When you select this item or click the button the current protocol information is saved Print When you select this item or click the button a dialogue opens up for you to select the printer type and other details of the printout Print preview This item allows you to see what will be printed out You can select whether you want a side by side two page view or a single page view You can also zoom in magnify the page to see more details alternatively you can zoom out reduce the magnification if the page is already magnified You can step to the next page or back to the previous one When you have finished viewing the pages that are going to be printed out you can click the Print button to get the printout or Close to return to the protocol editor Print setup You can define the printer type that will be selected You can also set the paper size and orientation Exit This closes the protocol editor If the protoc
75. es In the Instrument tab of Miscellaneous settings when you select properties of an instrument the nickname you selected appears and can be changed Also the official name and product number of the instrument as well as its serial number will be displayed You cannot change these Options This can only be accessed if you have Advanced level selected There can be four tabs but only three show on Advanced level the fourth requires Service level In the enhanced security mode this can be accessed if you belong to the Service user group All the four tabs plus the Automatic logout tab are shown 127 Tools menu General Options E General Instrument hardware Measurement technologies PerkinElmer 2030 assay result options Maximum number of assay results to be stored in the database 100 Assay results per protocol PerkinElmer 2030 Manager options TT Start all assay runs with Start Wizard PerkinElmer 2030 options 7 Run Workstation in demo mode no instrument needed Barcode usage mode Barcode is stored to results Platemap usage mode Assay ends after last plate in the plate map X Upon assay completion a new assay measurement is automatically started using the same protocol 7 Use Cw lamp filter Wheel B with 8 slots Cancel Help You can define the number of results to be held in the database for one protocol When this number is exceeded the oldest results for that protocol will be automa
76. es you the number of wells in the plate Number of well columns This is the number of wells in a column The product of this with the number of well rows gives you the number of wells in the plate Height of the plate This defines the height of the plate from its base to its top The units are millimetres 126 Tools menu Strip orientation You select whether the strips are aligned horizontally i e in rows so that the first strip enters the instrument first or vertically i e in columns so that first well of each of the strips enters the instrument first The normal setting is Horizontal The order of the measurement is according to the strip definition Plate dimensions The picture shows what dimensions are needed to define a plate The units are millimetres and you can specify the dimension to the nearest hundredth of a millimetre Note The distances should be measured from the outermost part of the plate Tune When you click the Tune button the Plate dimension wizard is started See the description starting on page 108 for how this works Well names In the dialogue that appears you can give a name for a well for identification purposes e g standard unknown etc You can define a short form single letter for the name and select a colour for it from a scroll bar of colours The short form is checked to see that it is not used for any other well name The default names are Empty and Measured Instrument properti
77. for the samples you are measuring so that you get the greatest variation in sample colours 20 Operating PerkinElmer 2030 Scale e Logarithm C Linear You can choose whether you want the scale to be logarithmic which means that the difference between smaller values will be more visible whereas the difference between higher values will be less visible or a linear scale in which every part of the range is treated equally In the enhanced security mode these controls are enabled only for users who belong to the Operator user group Kinetic Chart The Kinetic Chart showsa real time kinetic chart of the current well being measured The Y axis scale type can be changed between Logarithm and Linear If Scale to max is checked the Y axis maximum is linked to the maximum of the well repeat measurements 21 Operating PerkinElmer 2030 eleng ale el ciel Live display Kinetic Chart Temperature Type C Logarithm i Iw Scale to max Well repeat Measurement information Measurement technology TR fluorometry Latest result from Plate 1 repeat 1 label 1 well 401 well repeat 14 74203 Measuring assay 6 using protocol Kinetic For Help press F1 Temp C N A DEMO 22 Operating PerkinElmer 2030 Temperature e Kc PerkinElmer 2030 Manager Lie Eile Instrument Tools Help ale elel elei Live display Kinetic Chart Tempera
78. from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory pre set CW lamp filter names begin with a letter indicating the technology for which they are to be used P is for photometry After the letter comes the wavelength Excitation aperture PerkinElmer 2030 0050 only There are three sizes of excitation aperture controlling the size of the incident light beam You can select with the option buttons Small Normal or Large Normal is the default Reading time This parameter determines how long the sample is read to determine the absorbance Second measurement In the case where you want to make measurements with two filters you can click this check box and then set the parameter CW Lamp filter It has the same options as the parameter with the same name above When you select this two measurements are made the first with the normal settings and the second with this new setting The change is automatic 85 Tools menu Luminometry Luminescence in the sample is detected The parameters that can be set for luminometry are show in the picture and described below Luminometry Label Properties Name CPS Password Last Edited No user 2 15 2008 1 40 40 PM Parameters Emission Filter Nofiter SltA7 y Emission Aperture Ze Normal Small Large Counting Time 11 6008 Second Measurement Emission Filter No filter Slora z Y Canc
79. hat selecting medium instead of short approximately doubles the excitation energy for one measurement i e the number of flashes for one measurement doubles hence the time used for one measurement doubles and the number of counts doubles Selecting long instead of short triples the excitation energy the measurement time and the number of counts The number of flashes can be seen from the result viewer by selecting View all result data Note Each flash contributes to the total excitation energy Flashes keep on occurring until the required total excitation energy is achieved However because the energy comes in discrete flashes the last flash will cause the total excitation energy to exceed the required amount by a maximum of not more than the energy in one flash This will not be significant if there are many flashes but could be significant if there are only a few flashes E g in standard measurements the amount of flashes is about 1000 the variation in the total excitation energy is thus 1 1000 or 0 1 If the amount of flashes is about 100 the variation in excitation energies is 1 100 or 1 92 Tools menu Light Exposure Reference Level These values change the excitation energy approximately linearly Changing this value from 50 to 100 has about the same effect as changing the light exposure time from short to medium The number of flashes can be seen from the result viewer by selecting View all re
80. he Flush command again Note It may be necessary to use a solution of 70 ethanol rather than water for rinsing the tubing See the reagent manufacturer s instructions Needle maintenance Every time the dispenser has been used the tubing must be flushed properly When this is done the needles are rinsed at the same time However it is good to regularly clean the outside of the needles Avoid touching the needles with your fingers To clean the needles use e g a stick with a tip of soft material dipped in ethanol or use a spray bottle filled with ethanol Be very gentle when touching the needles so that they are not bent out of shape Rinse with water The position of the needles is very critical for high quality performance If a needle is bent call the service engineer to get the needle replaced Do not try to re adjust the damaged needle because the quality of dispensing could be affected Both the needle and the tubing will be replaced as a set It is good to check the flow of liquid dispensed during flushing If the flow is not normal it might indicate that the needle is partly blocked If this has happened due to dried reagents use water and e g ethanol to dissolve the plug If that does not help the needle needs to be changed contact the service 153 Appendix 4 Enhanced security special functions Appendix 4 Enhanced security special functions Note Enhanced security mode must be selected during installation You mu
81. here Notes This tab allows you to see any notes that are connected with the results being viewed 69 Result viewing Error Error gives you information about the general error situation and specific information about any errors that occurred for each measurement type Print Clicking this icon leads to an extended print dialogue which in addition to the normal print options gives you a choice of printing selected sheets or the entire workbook The former is the default You can also click the Preview button to get a preview of the pages to be printed In the case of high density plates the results from the first columns 1 to 18 or 1 to 24 appear on one page and the rest 19 36 or 25 to 48 appear on the next page Only rows of one plate will be printed on one page If you press OK in the print dialogue the selected parts of the run information will be printed on the printer that has been selected in the print dialogue Export results If you select Export in Result Viewer it allows you to export the results of the active plate A window appears in which you can give the path the file name type it in the file type select it from the list a Save button and a Cancel button The file types you can save files in are Excel 5 0 or later Tab delimited text or MultiCalc To help you select where you want the file to be saved you can browse to find the folder you want Buttons allow you to go up one level create a new folder and
82. in Administrators Administratars 2030Admin Edit User Password for 2030Admin Hidden Hidden 2030Admin Set Security Type Standard ES 20304dmin Register New Application N A PerkinElmer2030 2030Admin s H gt Export Clear Audit 2 fig OK Cancel Clicking the Protocol button launches a dialogue that displays protocol parameters from the selected audit trail entry The Protocol button is enabled when the audit trail entry is selected and it contains protocol parameters Clicking the Archive button launches the Archive dialogue similar to the Save As dialogue where the user can enter the path and filename of the archive text file The Save command archives the audit trail and deletes the archived entries from the database Only entries that are as old as or older than the selected date are archived Clicking the Export button launches the Export dialogue similar to the Save As dialogue where the user can enter the path and filename of the export text file The Save command exports all audit trail entries to the selected text file 159 Appendix 4 Enhanced security special functions Archive Note Archive appears only when the enhanced security mode is installed Archive is enabled only for users who belong to the Service user group Archiving deletes results from the database and archives the deleted results as text files The protocol parameters that were used with the assay run are also archived wi
83. ion Wizard This level allows editing of all system S a parameters Improper use of this mode Dispenser Maintenance can cause PerkinElmer 2030 system to behave unexpectedly User Level Options Cancel If the User Level is selected to be Routine this will automatically disable any system operations To enable these operations you must select the Advanced level Service is only for qualified service specialists It gives access to e g Instrument Options 74 Tools menu Labels Measurement Technologies In the enhanced security mode this tool is enabled only for users who belong to the Editor user group Labels TRF Fluorometiy Photometry Luminometry FP LANCE Europium Samarium Terbium Dysprosium Europium Fast Samarium in europium window LANCE 615 non VICTORY label LANCE 665 non VICTORY label Help In this window there are tabs corresponding to the measurement technologies labels in the instrument Each tab includes the names of the currently defined labels Names of user defined labels can be freely defined by the user but names of factory pre set labels cannot be changed See the sections below for details There are four buttons at the bottom of the labels window Add add a new item to the list Give the name and edit the properties Copy select an item to activate this Make a copy of an item It will appear with a new ic
84. it into a wheel positioned in front of the CW lamp There are eight positions on this wheel and two are reserved by default for fluorometry and three for photometry The remaining positions can have filters added by the user Note Default filters can be removed and replaced by user filters There are also two optional filter wheels filter wheel B and filter wheel B2 Filter wheel Bdeft side figure has four positions for the most common size of commercial filters i e round one inch Filter wheel B2 right hand figure has 8 positions for filters with a diameter of 15 mm Both wheels are supplied empty but with locking rings The system recognizes automatically if the type of wheel is A or B However if a B2 wheel is used instead of the default type B you need to inform the system through the workstation software This is done by selecting Use CW lamp filter wheel B with 8 slots in Tools Options General 102 General Instrument hardware Measurement technologies ee E r PerkinElmer 2030 assay result options Maximum number of assay results to be stored in the database 100 Assay results per protocol r PerkinElmer 2030 Manager options 7 Start all assay runs with Start Wizard r PerkinElmer 2030 options I Run Workstation in demo mode no instrument needed Barcode usage mode Barcode is stored to results Platemap usage mode Assay ends after last plate in the plate map
85. lide Emission filter and CW lamp excitation filter The former is a strip and the latter a wheel The instrument is loaded with slide A in each case but an empty slide B is also available for user filters as described below 99 Tools menu Emission filter slide e Emission filter stide A Properties Available unused Filters Filter Positions Al filters can be dragged from the Available Filters list to an empty position in the slide or from one position to an empty one or back to the list 020000000 MM Factory Defined Eject Slide W User Defined coca ze There are 8 positions in emission filter slides A and B The physical dimensions of the filter slots are diameter 25 5 mm depth 11 mm clear aperture 22 mm All standard filters with diameter of 25 4 mm 1 inch and thickness of 10 mm fit in the emission filter slides In emission filter slide A positions 1 2 and 8 are used for emission filters for time resolved fluorescent chelates if this option is included position 5 and 6 for emission filters for fluorometry labels umbelliferone and fluorescein respectively Position 7 is kept open for luminometry measurement If you want to add you own filters you must add them to empty positions or either use slide B which is fully customizable or change some of the existing filters Factory pre set filter names begin with a letter indicating the technology D is for Time resolved fluoro
86. ly You can find this program by clicking the PerkinElmer 2030 icon in your Windows Start menu It is called PerkinElmer 2030 Database maintenance Check that no other major resource consuming programs are running while using the PerkinElmer 2030 software Check that there is at least 30 MB available on your C drive Defragment your hard disk You can find this function in your Windows Start menu under Programs Acessories System tools Disk Defragmenter Processing error This dialogue will be displayed if the Instrument server goes to an error state or if the Manager cannot communicate with the Instrument server Right after the header there is text telling to user that Instrument server has stopped processing of the Workstation software because of an error After that there is a description about the error given by Instrument server or explanation that Manager cannot communicate with the Instrument server if that is the case When you click the OK button the reboot sequence is activated and the PC will be restarted System requirements not met This dialogue will be displayed during Manager startup if NoSystemCheckAtStartup command line option was not used and the minimum system requirements for Manager were not fulfilled The Current System panel is used to display information about the current system parameters The System requirements panel is used to display the minimum system parameters for the Manager to star
87. m size e Normal C Narrow I Second Measurement Emission Filter Cancel Help Label name This is the name by which the label is identified A password can be associated with it to prevent parameters being changed by an unauthorized person Flash Energy Area There are two options for this parameter Low and High The energy of the flash lamp used is dependent on the discharge capacitor value The Low Flash Energy Area selects a basic capacitor of 10 nF The High Flash Energy Area selects a second 10 nF capacitor in 77 Tools menu the lamp s discharge circuit thus doubling the energy If you select High you will reduce the counting time number of flashes but the counts will remain about the same Flash Energy Level The Flash Energy Level sets the voltage of the discharge capacitor the range is from 1 to 255 corresponding to 500 to 800 volts Note that although the lamp input energy depends on the squared power of the discharge voltage the output optical power is not linearly dependent on the input energy By changing this parameter you will fine tune the Flash Energy If you increase the number you will reduce counting time but counts from your sample will remain about the same Excitation Filter The non changeable filter in the flash excitation path D340 has a maximum transmittance at 340 nm and half bandwidth of about 35 nm The other two filters are only used for UV
88. mer I Ca Fluorometry EA Luminometry 1 GQ Photometry E Lei Time Resolved Fluorometry DELFIA m Eg Fluorescence polarization o L Time Resolved Fluorometry LANCE E L Assay examples L Luminescence Assays 1 Gq Bradford Protein Assays E Ca DELFIA Assays m TruPoint Assays a G LANCE Assays Aequorin Assays it ol x EN E lt Back Cancel Finish Definition of the sample names and number of plates See Samples tab on page 50 for a description of the principles involved in sample definition _ PerkinElmer 2030 Start Wizard Please specify number of plates and measured wells vlalrln e K l Plate 1 of 1 35 wells TE e 7 e e fol A 000000 B 000000 c 000000 a 666666 E 000000 F 000000 o 99999 H 000000 Note Errors in protocol selection or plate layout definition will lead to incorrect results 30 Operating PerkinElmer 2030 Comments that you want added to the results just type in what you want PerkinElmer 2030 Start Wizard Please insert free form text description that will be added to results lt Back cI Load the first plate the protocol name and comments appear in order to confirm that it is the one you want Note The Al position must be in the left corner furthest from the user i e the side that enters the reader first If
89. metry including LANCE Fis for fluorometry P is for photometry not used for emission filters L is for luminometry After the letter comes the wavelength 100 Tools menu User defined filters can be named freely but there is only space for five letters to be shown in the figure of the filter position The full name will be shown if you move the mouse cursor onto the slot where the filter is Physically changing the emission filter slide Click the Eject slide button on the Emission filter slide dialogue A message will appear telling you to unscrew and remove the plate on the side of the reader When you have removed the plate click OK to cause the slide to be ejected Pull the slide out carefully Avoid touching the filters with your fingers When you have got the slide out you can then change the filters or load filters into emission filter slide B When the slide is ready carefully return the slide to its position Push it in as far as it will go Replace the filter slide cover and screw it in place then click OK on the dialogue on the screen After this you can operate the instrument 101 Tools menu CW Lamp filter wheel CW lamp filter wheel A Properties Available unused Filters Filter Positions All filters can be dragged from the Available Filters list to an empty position in the slide or from one position to an empty one or back to the list WB Factory Defined HB User Defined Excitation filters f
90. mount of flashes is about 100 the variation in excitation energies is 1 100 or 1 78 Tools menu Light Exposure Reference Level These values change the excitation energy approximately linearly Changing this value from 50 to 100 has about the same effect as changing the light exposure time from short to medium The number of flashes can be seen from the result viewer by selecting View all result data Emission filters You can select the emission filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory pre set emission filter names begin with a letter indicating the technology for which they are to be used D is for time resolved fluorometry After the letter comes the wavelength Emission aperture In front of the photomultiplier tube there is a 4 position aperture slide which has three different apertures and a shutter position The normal aperture is circular with a diameter of about 4 mm and it is used for factory set labels The small aperture is circular but only 1 mm in diameter This allows you to shift the dynamic range of the emission signal for a particular user selected label which may otherwise exceed the linear range of the instrument This aperture is used for the top fluorometry factory set labels There is also a large circular aperture 5 mm in diameter The shutter is used at all times when no measurement is occurring to prevent str
91. n be the number of the protocol to be used for measuring that plate This system is especially useful for models using stackers In this case the first plate to be loaded from the stacker must have a barcode Measurement can be started simply by briefly pressing the button on the reader See page 118 for more details on barcode usage Plate orientation The figure shows a typical 96 well plate and how it is to be orientated when loading it either directly into PerkinElmer 2030 or into an input stacker The A1 position must be in the left corner furthest from you i e the side that enters the reader first If a barcode is used it should also be on the side that enters the reader first The barcode can then be used to identify the plate or to select the protocol 24 Operating PerkinElmer 2030 Loading direction Position Al Plates used in a stacker must have this ledge at least on the short sides Starting operation with the Start button 1 Open the loading position lid 25 Operating PerkinElmer 2030 2 Load your first plate and close the lid See the picture on page 20 for correct plate orientation 26 Operating PerkinElmer 2030 You can follow the measurement process by selecting the Live display Live display Kinetic Chart Temperature Protocol ATPlite 5 When the plate has been measured remove it and load the next plate 1 Remove the sure plate 2 Loadthe next plate
92. n click the Second measurement check box and then set this emission filter It has the same options as the normal emission filter When you select this two measurements are made the first with the normal settings and the second with this new setting The change is automatic 87 Tools menu Fluorescence polarization PerkinElmer 2030 0050 only Fluorescence polarization light of a particular wavelength is selected with a filter from the spectrum of a continuous wave lamp This light is then polarized and used to excite the fluorochrome in the sample The emission light is then viewed through two polarizers one parallel to the incident polarization S plane and one perpendicular to it P plane These are each in turn moved automatically into the emission light path The ratio 1000 Parallel G Perpendicular Parallel G Perpendicular is calculated to get the polarization in units of mP G is a correction factor see the end of this section The parameters that can be set for fluorescence polarization are the following Fluorescence polarization Label Properties Name FP Fluorescein D Del Password Last Edited No user 2 15 2008 1 41 21 PM Parameters DA Lamp Energy 65535 KREE D Lamp Filter Fa85 SlotAs sl Polarizer Aperture C Small Normal Emission Filter F535 SitA5 y Emission Aperture Ze Normal C Small Large Counting Time 1 01 6008 G factor H Cancel Help Label name
93. n the workstation interrupts the measuring process immediately The plate being measured is brought out to the loading position You cannot resume measurement after clicking Stop but must begin the assay from the beginning if required ie 39 Operating PerkinElmer 2030 Button on PerkinElmer 2030 This button has different functions depending on how long you press it for and what is happening when you press it If a plate has been measured you can remove it load a new plate and briefly press this button to continue the assay with the same protocol as the plate just measured However if you press this button for more than 2 s the current assay will be stopped and a new assay will be started with the current protocol This will not work if the protocol barcode option is selected If a plate is being measured and you press this button nothing happens If no assay is running and you press the button for more than 2 s then a new assay will be started with the current protocol This will not work if the protocol barcode option is selected or the enhanced security mode is installed Note Make sure in all cases that you have loaded the correct plate before pressing the button because the plate in the loading position or the empty plate holder will be taken into the reader Light on the button The following table shows you what the light on the button will be like depending on what is happening with PerkinElmer 2030 and whether or not the
94. natively once results have been received and stored in the computer you can view them by means of the PerkinElmer 2030 Explorer See the picture on the next page The protocol connected to the latest results will be selected automatically but you can select any protocol that was used to make the measurements that you want to view The results files associated with the protocol will then appear on the right side of the window Each entry for a results file comprises the following items Assay ID the date and time when the assay began and when it ended Notes and Errors Note To change a column width in the Results Viewer move the mouse cursor to the line between two columns The cursor form will change to a vertical bar with two arrows Hold down the left mouse button and move the column division in the direction you choose The information in a column is complete if no are shown at the end 65 Result viewing Click on the result file and then click the Open icon Ea gt e ajaja el Note These results are in the form of raw numerical data The Result Viewer gives details of the actual measurement There are five tabs and these described below The Result Viewer also provides you the following functions by means of command line menus or in most cases buttons File Export Button Page setup Print Button and Exit View List Button Plate Button Protocol Button Errors Button Notes Button View
95. nfirmation page Clicking Cancel closes the wizard and the normalization factors are not recorded in the database 115 Tools menu Normalization confirmation page PerkinElmer 2030 TRF LANCE Normalization Wizard The normalization measurement is now completed Check the results and press Finish to accept the new normalization parameters or Cancel to ignore them Normalization parameters Blank 482868 Cross talk ratio 0 0439 High amp 493443 High D 831153 SE Help This is the last page of the LANCE normalization wizard and appears after normalization has been successfully completed It shows you the measured normalization values and requires you to confirm that you accept these values before they are stored in the database The values obtained are the blank measurement in the acceptor window crosstalk i e the blank corrected contribution of the donor to the acceptor window signal as a proportion of the donor signal the maximum acceptor signal in the acceptor window and the maximum donor signal in the donor window These values are then used in a measuring sample to obtain the actual acceptor value i e the blank and crosstalk corrected acceptor signal normalized to the maximum donor signal thus eliminating quenching effects See the Calculation methods in the instrument manual for more details Note To avoid accidental clicking of the Finish button it only becomes visible after fiv
96. of the photomultiplier tube there is a 4 position aperture slide which has three different apertures and a shutter position The normal aperture which is circular with a diameter of about 4 mm should be used for fluorescence polarization Counting time This parameter determines how long the excitation continues The following times are recommended for 96 and 384 well plates Concentration nM Measuring time s 2 or more 0 1 1 0 2 0 5 0 3 0 1 1 0 89 Tools menu G factor In fluorescence polarization assays a G factor is used to correct for the bias in polarization values caused by differences in instruments and assay conditions To calculate the G factor prepare buffer and 1 nM fluorescein samples replicates should be at least 24 The formula used is ae G CorrectedS 1000 CorrectedP d L 1000 where the corrected parallel and perpendicular S and P counts are background subtracted fluorescence count rates obtained from a pure fluorophore solution e g CorrectedS averageS 1nM averageS buffer and CorrectedP averageP 1nM averageP buffer L is the literature polarization value for the fluorophore in mP E g the value for 1 nM fluorescein is 27 mP in aqueous buffer at room temperature The G factor is typically between 0 8 and 1 2 The G factor is used in the calculation of the corrected mP for samples as CorrectedS G x CorrectedP CorrectedS G x CorrectedP mP
97. ol has been changed it will prompt you to save it first Tools menu The Tools command line menu has one item Options When you click this a dialogue with a check box appears If you do not check it then whenever you click the Start icon measurement of the selected protocol will begin directly If you do check it then whenever you click the Start icon the Start Wizard will be used to start the measurement of the selected protocol In the enhanced security mode the Options menu item is enabled only for users who belong to the Service user group 61 Protocol editing 62 Result viewing 63 64 Result viewing Latest results The results from the latest assay can be viewed by clicking the Latest assay result icon PerkinEtmer 2030 Manager Eile Instrument Tools Help alel ey Ol In the enhanced security mode you must belong to the Operator user group to view the results of the latest assay The Result Viewer is started and the latest results shown after the measurement has been completed This command performs following actions 1 Tf there are no assay runs in the database yet a warning is given and the Result Viewer is not invoked 2 If this button is clicked while an assay run is already in progress that assay is displayed in the Viewer 3 If this button is clicked and there is no current assay running the latest assay run is displayed on the Viewer Results in the Explorer Alter
98. on showing it is different from a default item and the name will be preceded by the words Copy of You can give it a different name and edit other parameters by selecting Properties Remove select a user created item to activate this Remove the selected item from the list Preset items and labels used in protocols cannot be removed This command requires confirmation because you cannot undo Remove 75 Tools menu Properties select an item to activate this View the properties of the selected item If the item is user defined then you can edit the properties but if it is a factory pre set you cannot edit them 76 Tools menu Time resolved fluorometry Note To be enabled this requires the TR FIA option to be installed In time resolved fluorometry a flash lamp is used to illuminate the sample different lanthanides are used as labels The picture shows the parameters that can be set TR Fluorometry Label Properties Name Europium Password Last Edited Nouser 2 15 2008 1 32 58 PM Parameters Flash Energy Mode C Low High Flash Energy Level 245 E A ef Excitation Filter D340 C Optional C Optional Light Exposure Time C Shot e Medium C Long Light Exp Ref Level 157 ff Emission Filter D615 Slot 411 v Emission Aperture Normal C Small Large Counting Delay 1 400 2 j0 0 65515 us Counting Window 1 400 2 J0 0 65515 us Counting Cycle 1000 1000 65535 us Bea
99. or number There are also unique internal identifiers that the system gives for each protocol and assay run Keywords The following keywords can be used for file names in the protocol editor Outputs tab and in Assay end system command in the Events tab lt ProtocolName gt the name of the protocol Specified in the ID tab in the protocol name edit box lt ProtocolNumber gt the protocol number for the protocol Specified on the ID tab in the protocol number box lt ProtocolID gt a unique numeric identifier for the protocol lt AssayID gt a unique numeric identifier for the assay run lt RunID gt a numeric value identifying the specific run of a certain protocol 59 Protocol editing lt RunDate gt date when the assay was run in the format YYYYMMDD lt Plate barcode gt the barcode of the first plate of the assay If there is no barcode on a plate you should define an explicit file name for the plate lt Output file name gt the name of the output file given in the Outputs tab In some cases the numeric value of the AssayID or the RunID may not be suitable for the program handling the file You can add a function pad to allow you to decrease or increase the number You can specify lt AssayIDpadX gt where X 1 7 or lt RunIDpadX gt where X 1 4 Examples If lt AssayID gt 1234 and you add pad 2 to get lt AssayIDpad2 gt the result will be 34 This would be useful with MultiCalc which could not acc
100. own in the plate indicator text after the plate order number When you have selected the wells and released the left mouse button a drop down menu of sample types will appear These will be the types defined in the PerkinElmer 2030 Manager Tools menu item Miscellaneous settings By selecting one of these types you specify what the type of the selected area is to be Plate repeat parameters The number of times each plate is to be measured 1 300 If this is more than 1 then you must give the time between the end of one measurement and the beginning of the next 0 3600 secs This allows you to do slow kinetic measurements Measurement height Standard This selection is only enabled when using a plate type that has its dimension data stored in the instrument The default height set is about 8 mm above the bottom of the plate User defined If you do not want to use the standard measurement you can give anew value here You must click the user defined button The minimum value is 3 0 mm 8 0 mm if the dispenser is used and the maximum 18 0 mm depending on the plate type and if the dispenser is used or not The height is measured from the bottom of the plate This is useful if you are measuring samples on filter paper and you need to optimise for the sample being at the top or at the bottom of the wells Plate type Select the one you want from the list of available plate types under the scroll bar If the plate size and or format
101. oxes under the respective pump pictures The operation to be performed can be either Fill Empty or Flush and you can select which of these you wish to perform The operation is then performed on all the selected pumps Fill is used for filling the syringe and tubing before operation begins 150 Appendix 3 Dispenser operation Empty is used after operation ends to empty the contents of the syringe and tubing back into the bottle Flush is used for cleaning the syringe and tubing after it has been emptied Note Before performing this operations make sure the tubing from each dispenser has been put into water as described earlier Click Flush Take the tubing out from the container of water Empty any water remaining in the tubing by clicking Fill This draws air into the tubing and expels any water drops Fit the end of the tubing into the reagent bottle Click Fill to fill the tubing with reagent 151 Appendix 3 Dispenser operation Measurement Close the covers of the dispenser The system should now be ready for operation Proceed with the measurements in the usual way 152 Appendix 3 Dispenser operation Maintenance After measurement has been completed perform the following maintenance operations Click the Empty command This will cause the reagent to be sucked back into the reagent bottle so that none is wasted Remove the tubing from the reagent bottle and put it into a container of water and select t
102. r changing 101 Slide aperture 79 83 86 89 93 Slowdown in program 139 Software installation 13 Stabilized energy mode 82 Stacker 34 36 38 146 Start button 25 26 37 Start Wizard 29 37 61 73 128 129 Startup With enhanced security mode 17 Without enhanced security mode 17 Stop button 39 Strip orientation 127 System requirements not met 139 T Temperature 11 17 23 52 68 Temperature tab 23 Time resolved fluorometry picture 77 Tools menu 73 In main window 14 Troubleshooting 139 167 Index Tubing 149 Tune Plate dimension 127 U Up button Measurement tab 55 Usage of MultiCalc 144 User groups and permissions 154 User level 73 74 User management 154 155 V W Warning 139 Waste vial 148 Well columns 126 Well name Miscellaneous settings 127 Well names 124 Well operations 50 Well rows 126 View results 65 66 Wizard Dual label normalization 107
103. sssnassoscoveeseassbassessesvedacouscesncessensosnasonseseadoedesen 137 Performance degr dati n 2 cit edd EeeEet eeh EE aset 139 Processing CO ride Beie Seed tee 139 System requirements NOt met 139 Appendices engeren erden 141 Appendix 1 Connecting MultiCalc to PerkinElmer 2030 143 gr tee 143 MultiCale ssay protocols iii erben men bt tees 143 Installation of PerkinElmer 2030 output option in MultiCale 0 eee eeeeeeereeeeenees 144 MultiCalc assay protocols nono cnn n on nrnn ies 144 Usage of MultiCalc uma 2882 ans Eeer 144 Appendix 2 Plate types for stacker oooooccoonnonoconononnonnnononacnncononononcnnnonnonncncn conocia neraninnone 145 Wall e 2 22 422er aba ana an ed Hehe 145 Ne eu RO 145 VE 145 O Ee EE eet 145 Appendix 3 Dispenser Operation ccesesesesecseeeecseeeeceseeeesecseesecnersecnaeeecsaecaeesesaeeseens 147 Introduction e E 147 Emptyine the waste Val ptes lat er 148 Preparing the tubing its a ia Een 149 Dispenser maintenance rierien n n NEEN ee EEN 150 MEA 840688 Eed Bin IER ERAN 152 A EE 153 Needle maintenance i 4 hcg era ant an ine 153 Appendix 4 Enhanced security special functions useseessesnersesnersesnnesennenennsnennennnn 155 User groups and Permissions nennen 155 User Management 2 112 385 2482 8msm 8llssas ih E E R E E aR 156 Password Controls einn a E E e E E E E E dis 157 Auditado oe aap eae ee aE has 158 Archicad 160 Index cerir io E EE ENEE E N T EN E 161 I
104. st select the Custom setup option instead of the Typical setup See the Installation instructions in the Instrument manual for further information User groups and permissions The enhanced security mode usage of PerkinElmer 2030 is based on user groups and their permissions There are five different user groups and each group contains permanent permissions Groups and permissions are created during the installation and they cannot be added removed or modified Users can belong to several groups to get more permissions The available groups and their general permissions are e Administrators o User management o Change the security settings e Service o Archive the protocols and results o Change the general options e Auditor o View archive and export audit trail e Editor o Edit protocols o Edit labels o Edit filters o Edit miscellaneous settings o Run normalizations o Modify plate dimensions e Operator o Run protocols o View results of latest assay run o Use Dispenser Maintenance Note The following enhanced security functions can be found in the Tools menu 155 Appendix 4 Enhanced security special functions User Management Note User Management is only enabled for users who belong to the Administrators user group With User Management the PerkinElmer 2030 administrator can add remove and edit users and manage the password control Click the Add button to add users to PerkinElmer 2030 The New User
105. sult data Emission filter You can select the emission filter from the drop down list of available filters See the chapter on Filters for more information about how to define filters Factory pre set emission filter names begin with a letter indicating the technology for which they are to be used D is for time resolved fluorometry After the letter comes the wavelength Emission aperture In front of the photomultiplier tube there is a 4 position aperture slide which has three different apertures and a shutter position The normal aperture is circular with a diameter of about 4 mm and it is used for factory set labels The small aperture is circular but only 1 mm in diameter This allows you to shift the dynamic range of the emission signal for a particular user selected label which may otherwise exceed the linear range of the instrument This aperture is used for the top fluorometry factory set labels There is also a large circular aperture 5 mm in diameter The shutter is used at all times when no measurement is occurring to prevent stray light getting to the photomultiplier tube Counting parameters These determine the time factors involved in the measurement process all times are in microseconds Counting delay the time after the excitation pulse at which counting of the emission signal begins Counting window the time period within which counting occurs Counting cycle the time between excitation pulses within a measur
106. sured normalization values and requires you to confirm that you accept these values before they are stored in the database The two values produced by the normalization are the background and the europium contribution to the samarium window During actual dual label measurements the europium counts in the europium windows are first calculated by subtracting the background from the measured counts Knowing the europium counts and the normalization ratio the actual samarium counts in the samarium window can be calculated by subtracting the europium contribution to the measured counts Additional information about the reliability of the factors is also shown so that you can be sure that the results of the measurement are correct Note To avoid accidental clicking of the Finish button it only becomes visible after five seconds have elapsed since the page was opened During the time the Finish button is disabled the text of the button is Wait t where t is the remaining waiting time and it is updated every second When you click Finish the new values replace the old values in the database The Back button is enabled only if the wizard moved to this page from the Measurement warning page 111 Tools menu LANCE normalization Note TRF technology must be installed for this to be enabled In the enhanced security mode this tool is enabled only for users who belong to the Editor user group Using the LANCE normalization wizard Before LANC
107. t The Suggestions panel will list the reasons why the Manager cannot be started and a description about what must be done to fix the situation When the OK button is clicked the dialogue together with the manager is closed 139 Troubleshooting The minimum system requirements are Windows XP Intel Pentium Processor or equivalent 128 MB RAM or more 30 MB free hard disk space or more CD ROM Drive XGA Drive minimum setting is 1024 by 768 pixels Wallac Instrument Interface Board 140 Appendices 141 142 Appendix 1 Connecting MultiCalc to PerkinElmer 2030 Appendix 1 Connecting MultiCalc to PerkinElmer 2030 Introduction MultiCalc can read assay text files from PerkinElmer 2030 Evaluation of assays can be fully automated Measured wells are read sequentially Plate printout and re order work only for full plates MultiCalc version 2 6 or higher is required Note The Windows properties setting for MultiCalc should have Misc Background Always suspend NOT CHECKED Idle sensitivity Low To see what the situation is click the MultiCalc shortcut icon with the right mouse button and select Properties from the menu In the dialogue that appears select the Misc tab and then in the Background field make sure that Always suspend is not checked See also Installation in the Instrument Manual for how to install this option in MultiCalc MultiCalc assay protocols Create or edit a protocol using the Explorer an
108. t plate gives information directly relating to the label or result type displayed in the plate grid below it The information included is Name of the label or result type displayed in the result plate Unit for the results from the result plate Instrument Background value measured for the label or result type Protocol _ Assay 7 3 6 2008 1 26 33 PM PerkinElmer 2030 Result Viewer og Eile View Plate Help aja moe ele gt 2 Protocol description Protocol name vou Kinetic Protocol number NIA Name of the plate type Generic 8x12 size plate Number of repeats 1 Delay between repeats 08 Measurement height Default Protocol nates Kinetic repeats 50 Kinetic delay 00s Europium Time resolved fluorometry D615 Second window delay time Ops Second window time 0 pS Light integrator capacitors 2 Light integrator ref level 157 Flash energy area High Flash energy level 245 Flash absorbance measurement No Beam cc Normal 4 List Plates 1 1 A Plate Pagel A Protocol A Errors A Notes Ready Errors may have occurred in measurement Check results Selecting Protocol causes a list of protocol parameters to appear There is also a plate map showing which sample types were on the plate Any notes written in the protocol editor Notes box will appear
109. th the results The archive module shows the protocol tree in the left pane and results in the right pane as in the protocol explorer The User can select multiple results from the right pane for archiving Right clicking selected results brings up the Archive menu command the Archive command is found also from the Archive File menu The Archive command launches the Archive dialogue similar to the Save As dialogue where the user can enter path of the archive directory The Save command archives results and deletes the archived results from the database Each result is saved as a separate text file The archived files are named by protocol name and assay ID From the left pane the user can select protocols to be deleted Only protocols that have no results in the database can be deleted i e results must be archived first From the File menu the user can select the Archive old results command The command launches a dialogue where the user can enter the date Only results that are as old as or older than the selected date are archived The Archive button launches the Archive dialogue similar to the Save As dialogue where the user can enter the path of the archive directory The Save command archives results and deletes the archived results from the database 160 Index 161 162 Index 2 21 CFR Part 11 12 A Account lockout 157 Active protocols 26 Administrator 154 Advanced level 74 127 Aperture slide 79 83
110. the Operator user group When the workstation is used without an instrument i e running in demo mode the status bar includes the text DEMO When PerkinElmer 2030 workstation is connected to an instrument running in normal mode the demo mode indicator is blank You can switch between normal and demo modes using the Options selection in the Tools menu Note that you must restart your computer before this change will take effect 19 Operating PerkinElmer 2030 GPerkintmer 2030 Manager Jia g File Instrument Tools Help e pa ce Live display Kinetic Chart Temperature Protocol Absorbance 2 450 0 1 d L H Scale 11 Logarithm C Linear oma oma um ER soon vn a 004 em 1 0004 o amp r o 000 A Measurement information Measurement technology Photometry Latest result from Plate 1 repeat 1 label 1 well G01 well repeat 1 1 400 A Measuring assay 2 using protocol Absorbance 450 0 18 For Help press F1 Temp CO NIA DEMO You can choose the result range you want used to determine the colour code of the wells This scale utilizes rainbow colours so that the closer a result is to the upper limit of the range the more red it will appear The closer to the lower limit it is the more purple it will appear These colours will correspond to the values shown in the legend appearing beside the plate map Choose the range that is appropriate
111. thing in that area switch power off first N Caution Do not put your fingers into the sample loading area In the stacker 1 In stacker operation plates are loaded into the input stacker with the first plate to be measured at the bottom See the picture on page 20 for correct plate orientation 2 The empty output stacker the one with the tabs for the quick release mechanism should be fitted first into the multilabel reader it occupies the left hand position 3 Fit the input stacker with plates in the right hand position 1 Load the output stacker Release tab for the stacker 4 Select the protocol and click the Start button or use the Start Wizard 36 Operating PerkinElmer 2030 5 The plates will be moved one by one from the input stacker to the measurement position After each plate has been measured it is moved to the output stacker When the assay has been completed operation will stop If there are still plates in the input stacker it means that you must start a new assay by selecting the protocol and clicking Start as described above 6 When all plates have been moved to the output stacker it can be removed To do this press the release tab at the bottom of the stacker then lift it up After you have removed the stacker pull up the handles of the quick release mechanism to allow the plates to be emptied easily from the stacker Note Make sure you do not operate this mechanism when the stacker is loade
112. tically deleted from the database to make room for new ones The maximum number of results is 100 If you want the start to be always by means of the Start Wizard check this box This means that when you click Start the Start Wizard will be activated This is useful if e g you need to define the number of plates before each measurement If you just want to run the software as a demo without having the reader connected check the Run Workstation in demo mode check box Note If you change to Demo mode your PC has to be rebooted before the change takes effect Make sure you have no other programs running when you make this selection because they will be closed during the shutdown procedure If do not have the barcode reader option then the line Barcode reader switched off will be selected If you have the reader and want to use the barcode as a plate number to be 128 Tools menu connected with your results select Barcode is stored to results In both these cases you must select the protocol manually see Start or Start Wizard for more information If you have the reader and you select Barcode is used as a protocol number then the barcode will determine the protocol used to measure the plate If a plate does not have a barcode then the protocol used to measure the previous plate will be used See the Plate orientation picture on page 20 for where to fix the barcode You can specify what happens
113. tically if property A has been set as described above Evaluation can also be started manually by selecting COUNTER 1420_2 in MultiCalc and pressing Enter Re evaluation is also possible there after the Recover function has been used 144 Appendix 2 Plate types for stackers Appendix 2 Plate types for stackers All stackers are tested with following plates until further notice given Note Lids or loose frames can not be used in the stacker Wallac 1244 550 DELFIA microtitration strip plates C554 160 Rigid low fluorescence plate C120 105 Anti mouse coated plates C122 105 Streptavidin coated plates 1450 510 Rigid sample plate 1450 410 Rigid sample plate 1450 514 515 Isoplate Nunc 475515 FluoroNunc Black strip plate 437591 FluoroNunc White strip plate 45161 FluoroNunc Clear non strip 477178 8x 12 strip plate clear 242757 93 384 well white 242757 384 well clear Costar White clear bottom Black clear bottom Black White Greiner 655101 Clear non strip 655074 White non strip 655077 Black non strip 655180 Clear sterile Only without lids Falcon 3915 Clear non strip White non strip 145 Appendix 2 Plate types for stackers Labsystems 95029550 Combiplate white 9502817 Cliniplate black Acceptable range of plate dimensions for plates in a 1420 216 Stacker Length 127 2 128 2 mm Width 84 5 86 0 mm Height 14 0 25 0 mm Ledge height 1 5 6 5 mm Note The length and width incl
114. ties of an emission filter or you want to define the properties of a new filter a further dialogue appears where the name of the filter and the technology for its uses can be defined There is also a section where you can add additional information that you can connect with the filter properties Factory pre set emission filter names begin with a letter indicating the technology for which they are to be used D is for time resolved fluorometry LANCE filters are marked as such F is for fluorometry L is for luminescence After the letter comes the wavelength 97 Tools menu CW Lamp filters Filters Emission Filters d Filter Slides Close Help If you select properties of a CW lamp filter or you want to define the properties of a new excitation filter a further dialogue appears where the name of the filter and the technology for its use can be defined There is also a section where you can add additional information that you can connect with the filter properties Factory pre set CW lamp filter names begin with a letter indicating the technology for which they are to be use F is for fluorometry P is for photometry absorption After the letter comes the wavelength 98 Tools menu Filter slides Filters Emission Filters Cw Lamp Filters e Emission filter slide A Emission filter slide B Cw lamp filter wheel A Cw lamp filter wheel B There are two types of s
115. time resolved fluorometry LANCE homogeneous time resolved fluorometry 131 Tools menu 132 Exiting 133 134 Exiting File menu The File menu in the main window contains one item Exit This allows you to exit from the workstation software You need to confirm the operation If you try to exit while an assay run is taking place you will get an error message telling that this is not possible In the enhanced security mode the File menu also contains a Logout menu item This allows you to log out from the workstation software You need to confirm the operation After the logout the login dialogue appears Automatic logout Note Automatic logout tab appears only when the enhanced security mode is installed You can define the timeout before the user is automatically logged out Options E General Instrument hardware Measurement technologies Automatic logout Automatic logout options Timeout before automatic logout 1 60 15 H minutes The range for the timeout is 1 60 minutes The default value is 15 minutes 135 Exiting 136 Troubleshooting 137 138 Troubleshooting Performance degradation If you find your PerkinElmer 2030 software is not running as fast as it used to please run the PerkinElmer 2030 Database Maintenance program You should anyway do thison a regular basis This program will compact your database and will in some situations increase the software performance drastical
116. ture Plate heating On Off Target temperature 37 15 50 C Temperature TC 0 1210 1220 1230 1240 1250 1300 1310 1320 1330 1340 1350 14 00 Time 24 Hrs ____ A a For Help press Fi Temp C 37 4 In the Temperature tab of the main window you select plate heating and the target temperature The latter can be set with 0 1 C precision within the interval 15 50 C The minimum temperature is 15 C or room temperature plus 2 C depending on which is higher If the target is changed the new value is only effective when the Apply button has been clicked The figure shows the temperature rising rapidly to its new level and then stabilizing there The current temperature is shown in the bottom right hand corner of the window irrespective of which tab is selected 23 Operating PerkinElmer 2030 In the enhanced security mode these controls are enabled only for users who belong to the Operator user group Barcode options An optional barcode reader can be included This allows you to load barcode labelled plates which are identified by the barcode reader Codabar Code39 Interleaved 2 of 5 Code 128 UPC and EAN barcodes can be read There are two sorts of identification that can be used This identification can be by plate number in which case you must select the protocol as part of the start up procedure Alternatively the barcode ca
117. ty mode does not alone ensure compliance with 21 CFR Part 11 Any facility that wants to be compliant with the 21 CFR Part 11 regulation must also implement the necessary procedures and controls set by the regulation 21 CFR Part 11 The regulation 21 CFR Part 11 sets the criteria under which the Food and Drug Administration FDA of the USA considers Electronic Records and Electronic Signatures if applied to be trustworthy reliable and generally equivalent to traditional paper records and handwritten signatures executed on paper The regulation applies to records in electronic form that are created modified maintained archived retrieved or transmitted under any records requirements set forth in agency regulations i e GMP GLP GCP It also applies to electronic records submitted to FDA under requirements of the Federal Food Drug and Cosmetic Act Enhanced Security mode PerkinElmer 2030 has an Enhanced Security mode intended for facilities that have to comply with 21 CFR Part 11 regulation from the FDA During installation you can select if you want to use the Enhanced Security mode Once you have enabled this mode you cannot disable it because to do so would not be in compliance with 21 CFR Part 11 The Enhanced Security mode for PerkinElmer 2030 provides the technological controls and features to support full 21 CFR Part 11 compliancy These features can be classified under three main headings e improved access control unique
118. ude the size of the ledge 146 Appendix 3 Dispenser operation Appendix 3 Dispenser operation Introduction The optional dispenser allows you to make measurements that require e g the addition of reagent to start the process as in the case of flash luminescence or start and stop an enzyme reaction It can also be used for dispensing Enhancement Solution for a DELFIA time resolved fluorometry measurement It can be used with any of the technologies supported by PerkinElmer 2030 The dispenser can have up to four pumps fitted This allows you to have separate pumps dedicated for specific purposes All the needles are directed to the same well allowing dispensing of more than one reagent to the same well The number of pumps fitted and the total volume of each pump are set during installation This information then appears when dispenser maintenance is selected The default setting for the dispensed volume can be between 5 to 350 microlitres Note Dispensing to 384 well plates is possible if just one pump is installed The optional dispenser is a separate unit from PerkinElmer 2030 itself The information about installing the dispenser unit will be found in the Instrument manual as part of the installation instructions The following instructions tell how to use the dispenser 147 Appendix 3 Dispenser operation Emptying the waste vial Note The waste vial must be empty when operation starts To check that the waste vial is
119. ument Click Next to start the measurement of the plate Measurement status page PerkinEtmer 2030 TRF LANCE Normalization Wizard The measurement has now been started Measurement status Assay is running Measurement progress i Measurement in progress This is shown during the actual normalization measurement and it gives you feedback about the measurement The measurement status pane shows the current status of the measurement The progress of the measurement is shown by the progress bar 114 Tools menu O PerkinElmer 2030 TRF LANCE Normalization Wizard The measurement has now been started Measurement status Assay is ready Measurement progress Cancel Help The Next button becomes available when the measurement has been completed If the measurement was completed without any warnings the window shows the text Assay is ready Click Next to move to the Normalization confirmation page otherwise the wizard moves to the Measurement warning page Measurement warning page This is used to inform you that something out of the ordinary has happened during the measurement of the normalization assay This page gives you enough information to decide whether the results can be accepted regardless of the problem that caused the warnings Clicking Next on this page advances the wizard to the normalization co
120. ut is 10 Lockout duration defines the time spent after the maximum number of failed login attempts The Permanent until administrator unlocks option button means that the user will be unable to log in again until the administrator has assigned a new password The Duration option button prevents the user being able to login again until the time specified has elapsed Duration is greyed if Permanent is selected By default the Lockout duration is 60 minutes Audit Trail Note Audit Trail appears only when the enhanced security mode is installed Audit Trail is enabled only for users who belong to the Auditor user group 158 Appendix 4 Enhanced security special functions By using the Audit Trail tool you can view the operations performed by authorized PerkinElmer 2030 users The dialogue contains a list where all non archived audit trail entries are shown sorted by date User Management Users Password Control Login History Audit Trail Summary Function Previous Value Current Value Full Name Clear Login History N A NJA 2030Admin Change Login Type Windows Login PerkinElmer Login admin Edit User Groups for PERKINELMERN NJA Operator Editor admin Edit User Groups for LASSGMLDO9 a Administrators Administrators admin Load Windows Users for Windows Lo PKIUsers Administrators 2030Admin Change Login Type PerkinElmer Login Windows Login 2030Admin Edit User Groups for 20304dm
121. w plate delete the currently selected plate or duplicate a plate An added plate has all the positions marked as Measured and will become the last plate in the sequence When you delete a plate it is the one currently visible that is deleted A duplicated plate is a copy of the current plate and will become the last plate in the sequence A map of all the wells appears An empty well i e one that will not be measured will be grey a well with a sample i e one that will be measured will be coloured If you move the cursor onto a well the sample type will appear If you click on the well a list of available well types will appear To change the markings for the individual wells in a plate you can select a group of wells by using the mouse To start the selection click the left mouse button at the desired starting 50 Protocol editing position and then drag the mouse cursor to the desired ending position on the plate without releasing the left mouse button Complete rows or columns can be selected by clicking on the character number on the plate frame corresponding to the row column More than one row column can be selected by clicking the mouse and dragging in the same way as when selecting free well areas The whole plate can be selected by clicking on the upper left corner of the plate frame To select a rectangular area of wells on the plate hold down the Shift key while dragging with the mouse The number of wells selected is sh
122. y Level ida 92 Excitation filter z stos palas asias 92 Lisht Exposure 92 le 92 Light Exposure Reference Level 93 Emission UE 93 EMisSSIOM ApertllE tantito dada ets 93 Counting parameters cuidara 93 Factory set values for the main counting Window nen 94 J Eet oea ANN 94 IKEA 94 Delay EE 94 Window E 94 Window 2 nr rd nen 94 Cycles inn A rata 94 Bedia ii dde atte 94 Second measurement parameters 0 0 0 ee eeeeceseceseceseceseceecseeeseeeaeeeeeeeseeeeeeeenseenaes 94 LANGE Label Pr perties 2 43 04 24868 e Taan Eear EEOAE SEEE santos 94 Filters sn a niit nit 96 Emissi n filters e incest ec 97 CW Lamp oie ee Resets Shee bee 98 Filter slides ai A 99 Emission iter Sle oe risann rasen nme 100 Physically changing the emission filter sde 101 CW Lamp filter wheel Heer een ins 102 Contents Physically changing the CW Lamp filter wee 104 Changing the EW Eamp 2 22 sore petia ee Eeer e Ea eR Era a ea Ea aSa 104 Changing filters in the software 105 Filter propertiestss assent htc seenen Eeer 106 EuSm Dual label normalization oooocconononocnononnnonnononnnononncnnnononnnono cnn nnnon non cono no na ncrnninnone 107 Plate type Selection mii Ee deet BEE 107 Preparing the plate ur eege dek Age Eeer EES sr 109 Measurement status page 110 Measurement warning D ge nono nono nn non nonnonnon non n canon nc non nn ran rnnonncnno 110 Normalization confirmation page 111 LANCE normalizatio s cit ehe hase aa ER EREE
123. y preset emission filter names begin with a letter indicating the technology for which they are to be used F is for fluorometry After the letter comes the wavelength 82 Tools menu Emission aperture In front of the photomultiplier tube there is a 4 position aperture slide which has three different apertures and a shutter position The normal aperture is circular with a diameter of about 4 mm and it is used for factory set labels The small aperture is circular but only 1 mm in diameter This allows you to shift the dynamic range of the emission signal for a particular user selected label which may otherwise exceed the linear range of the instrument This aperture is used for the top fluorometry factory set labels There is also a large circular aperture 5 mm in diameter The shutter is used at all times when no measurement is occurring to prevent stray light getting to the photomultiplier tube Counter position The counter position can be selected to be Top or Bottom Counting time This parameter determines how long the excitation continues Second measurement parameters In the case where you want to make measurements with two filters and or two different energies you can set these parameters in this field Click the check box Second measurement to activate them There are three of them CW Lamp energy CW Lamp Filter and Emission Filter See the corresponding sections for the first measurement for more details When you
124. y used protocol 129 Tools menu Instrument hardware Options General Measurement technologies Instrument hardware options Stacker Installed Barcode reader Installed Temperature control Installed Adjustable beam size Installed Counting from below Installed High power flash driver Installed UY Absorbance Installed Dispenser Installed Pump 1 Installed Pump 2 Installed Pump 3 Installed Pump 4 Installed Cancel Help This tab can only be accessed if Advanced level has been selected in User Level In the enhanced security mode this tab can always be accessed It shows which options are installed These include Stacker option Barcode reader options Temperature control Adjustable beam size Counting from below High power flash driver Dispenser and pumps UV absorbance 130 Tools menu Measurement technologies Options General Instrument hardware r Available measurement technologies Fluorometry Installed and available FP Installed and available Luminometry Installed and available Photometry Installed and available TR fluorometry Installed and available LANCE Installed and available Cancel Help The final section of this tab tells what measurement technologies are installed and available Fluorometry FP Fluorescence polarization Luminometry Photometry TR Fluorometry dissociation enhanced
125. you want dispensing for every repeat then leave it unchecked Clicking the OK button inserts a dispense operation in the measurement operations list Delay button Clicking this causes a dialogue to appear that lets you specify the duration of the delay between the completion of the preceding operation and the start of the one succeeding the delay operation in the operation list You use a slide bar to do this Clicking the OK button inserts the delay operation into the measurement operations list A single delay can be from 0 1s to 3600 s At the bottom of the dialogue is a check box Perform on first plate repeat only If you only want delay for the first of the plate repeats check this box If you want delay for every repeat then leave it unchecked Shake button Clicking this causes a dialogue to appear that lets you specify the parameters for a shake operation You can specify the duration of the operation the speed slow normal or fast the extent the shaker moves and the path that it follows straight line elliptical or figure of eight linear orbital or double orbital respectively Clicking the OK button inserts the shake operation into the measurement operations list At the bottom of the dialogue is a check box Perform on first plate repeat only If you only want shaking for the first of the plate repeats check this box If you want shaking for every repeat then leave it unchecked Label button Opens the Label Sel

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