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1. 45 Using the Assay Start Wizard Select the technology to be used The name selected will appear at the bottom of the window Technologies marked with are ready to be used for measurements Click Next Note If you are using a protocol with multiple technologies you must run the Assay Start Wizard as many times as there are labels to optimize When the final technology is optimized then you can run the assay Optimization information Note This step in the wizard and the following ones are common to both the Create new protocol and Optimize existing protocol options Y Assay Start Wizard Optimization info Please select optimization s you want to run For advanced options check the Advanced made checkbox Click Next to continue Optimization info Optimizations to run Plate Dimension Not optimized Measurement Height Not optimized Detector Gain Not optimized z Not optimized ib h i Flatfield Correction Not optimized Advanced mode 96 General e Fluorescein lt Back Next gt Cancel 46 Using the Assay Start Wizard This shows you which optimizations are needed for the plate and technology combination you have chosen and if any of these optimizations have been done already You can select any or all of the first four optimizations The fifth Flatfield will be disabled because it cannot be done at the same time as any of these Assay Start Wizard Optimization inf
2. Result viewing Curve fitting standards on each plate Label Calculation Fluorescein 1 Channel 1 535 Fitting LinRegr v Show Info Frames Y axis transformation here E Edit Fitting Parameters i i aM Edit Display Parameters Y axis transformation None he Activate Inactivate Replicate Points Reject outliers O lick Advanced Parameters Ed Show Gridlines E p Calc Unknowns Concentration unit v STD Concentration LJ 1 5 i 2 500 320 o0 wee eee eee eee wee eee eee eee eee eee eee eee see eee eee ee on Show info frames an extra pane appears with more information about the curve including the slope the intercept the estimated dose at various percentages and the variance Assay J Standard curve fitting i Protocal General settings Aue Weib selection Group t Label Catcutation Piucrescein Channel 1 S25 a avoc toe Reg v Advanced Pas seontees sends transformation v Nore Toara tranformar None ws Reject outhers C ick Advanced Parameters Ede Fating Parameters Conemrts ater unt STD Concentration t s 2 w lt gt Rasuk vel be re calcuated nhan you press the Ansty burton 86 Result viewing Edit fitting parameters a dialog appears allowing you to set many parameters which affect the curve fitting Some of the parameters to do with curve fitting also appear in the main calculation window Fitting Options Cur
3. High scale froooood 1 000 000 800 000 600 000 400 000 200 000 fi This view has three tabs The first two Counts and Graph the same as Kinetics correspond to those described in Reader control Assay 7 Results File Edit View Tools Actions Help gt lo x Q 0 Back Forward Up on Group 1 a B Assay B Protocol A Notifications lt Plate 1 Group 1 ID Fluorescein 1 B B Calculations fe 1 Curve fitting star Assay ID 7 Simulated 2 avg of type 3 avg of type 81 View Plate Size to fit z ERES Result viewing The same features and side bar functions are available as in Reader control The third tab List shows results in the form of a list of numerical values Assay 7 Results loj x Eile Edit View Tools Actions Help 9 090 2 f Back Forward Up Export Cale Group 1 Assay ID 7 Simulated bond E H Assay Bp Protocol LD Notifications B s Plate 1 4 Group 1 ID Fluorescein 1 B Calculations 4 PlateID PlateNumber PlateRepeat GroupID Well Sample Pointy Point Labellndex Labelinst 2 Avg of type i 3 avgot type 8 1 1 1 AOL UNKI Q o 4000000 1 8 1 1 A02 UNKI o 0 4000000 1 8 1 1 1 A03 UNK2 o o 4000000 1 Note In Group view any saturated result will be highlighted with gray under the Counts tab 82 Result viewing Calculations 3 avge type aki
4. 479904 Click Next to go to the final window of the wizard 111 Advanced optimization 112 Chapter 5 Dispenser control 113 114 Dispenser Control Dispenser Control Click Dispenser Control to access functions and parameters involved in the operation of the dispenser There are two tabs Maintenance and Settings Maintenance You can perform maintenance operations with the dispenser by selecting the pump and the operation Initialization Select the pump or pumps to be initialized then click the Init button to perform initialization Wallac EnVision Manager 1 11 Simulation mode Fle Ede Vew Toss Actions Help 0 8 7 Bod Log Out Latest ihre Nat whikoed Initializing Press F1 for help Smmidetion mode aaa 115 Dispenser control Initialization resets the pumps by setting the valves and syringes to their home positions This may involve liquid being expelled through the aspiration tube into the liquid reservoir If you are not sure what liquid was used last direct the aspiration tube into an empty reservoir to avoid accidentally mixing liquids Note the other maintenance operations are disabled until Init has been clicked and initialization performed Rinse 2 OR Start Later Martenance Settings Iratiakuation Orro t trtistoed Puro 2 Intiskned Dispense s selected volume of kgad into the waste cortaner fill tubanga haga ate orcas hepa to fil the b
5. First Retrieve the reagent and then Fill the tubing 135 Dispenser control if rinse liquid has been left in the tubing then you have to get rid of this before dispensing reagent You can Retrieve the rinse liquid and then Fill the tubing Alternatively you can empty the tubing using Rinse without aspirating any liquid i e with the inlet tube in air Then replace the inlet tube in the reagent reservoir and Fill the tubing After operation Empty syringe then Retrieve reagent back into the reagent reservoir Replace this reservoir with one containing rinse liquid Rinse the tubing Either leave the tubing filled with rinse liquid or empty it i e Rinse with the inlet tube in air All these operations as well as others can be found under the Maintenance tab when you click Dispenser Control Changing a tip mount Note before changing a tip mount the tubing should be emptied You can do this by selecting Retrieve liquid or Rinse with the inlet tube in air To change a tip mount follow the instructions in the Routine maintenance part of the Instrument manual 136 Index 137 138 Index A Absorbance 3 Active protocols 68 73 Add curves 24 Advanced settings 145 AlphaScreen 3 Aperture 30 Aspiration speed 146 Aspiration tube volume 144 146 Tip tube volume 146 Total tube volume 146 Assay Results 85 Assay combinations 67 70 72 74 Assay Start Wizard 45 111 Asterisk 14 31 B Barcode Ru
6. Size of scanned area 56 TRF window 39 54 61 Wavelength 38 Z 40 63 Z target value 113 Optimization plate 45 65 Output 4 Overlaid curves 24 Remove all 26 Overview 3 P Pause button 69 71 73 75 Plate Loading 42 Optimization 45 65 Orientation 42 Results 87 Plate dim optimiz 37 55 111 Plate view 19 26 Prepare inst for transport 11 Protocol Results 85 Pump Connection to tip 146 PUMP button 128 131 134 137 139 141 R Reader control 19 Counts 19 Events 19 Kinetics 19 Temperature 19 Reader control window 10 Menu title bar 10 Toolbar 10 Relative to greatest 26 Remember settings 144 Remove curve 26 Result view Export 107 Results 81 Assay 85 Group 88 Notifications 87 Plate 87 Protocol 85 Refresh 82 Selection by amount 82 Selection by date and time 82 Selection by protocol 82 142 Results tree 85 87 88 Results window 84 Retrieve 147 Retrieve liquid 133 To empty tubing 148 Rinse 127 147 Aspiration tube volume 144 Show advanced options 142 Speed 144 Tip tube volume 144 To empty tubing 148 Total tube volume 144 Total volume 143 Run button 69 Run log 29 S Save Screenshot to File 13 Scan plate for strongest sample 56 Scanning 4 Settings 145 Advanced settings 145 Aspiration speed 146 Aspiration tube volume 146 Connected to tip 146 Heating 145 Set button 145 Stirring 145 Shaking 4 Shortcuts bar 16 Dele
7. Y a S Smax b S B S B max a 1 b 05 a 05 b 1 a 1 b 1 Choose between these three options to get the plot that most clearly helps you determine the window you want to use You can adjust the delay and window width manually if required Detector gain optimization FP and FI This optimization determines the optimum gain for the detector A single sample is required This sample should represent the strongest intensity in the assay Unless you choose otherwise the strongest sample will be identified and used in the case of fluorescence intensity For G factor optimization in fluorescence polarization a low binding or free fluorophore sample is needed The optimum gain is the one that is the maximum possible without saturating the detector for high signal samples Z optimization All technologies The optimization determines how well the sample results are separated from the background when the scatter of results is taken into account Six high samples and six lows samples are needed You can select the target Z factor the default is 0 6 In the results the minimum number of 37 Options for running EnVision flashes to achieve the Z target is selected You can adjust this flash number manually if required Flatfield correction This corrects for variation in the signal across a plate due to e g plate curvature In such a case the signal at the edges may be less than at the center due to the difference in dist
8. recently obtained Edit protocol edit the protocol shown in the drop down menu on the Reader control toolbar Run Assay start running an assay using the protocol shown in the drop down menu on the Reader control toolbar Introduction to the User Interface Pause this is only enabled if an assay is running You can temporarily stop the run and then continue it by clicking Run Note that with on the fly and HTS AlphaScreen measurements measuring continues until the end of the row before pausing Stop this is only enabled if an assay is running The run stops any results are saved and the instrument returns to the Idle state Unload Load the function available depends on if there is a plate in the instrument to be unloaded or if there is no plate in which case one can be loaded Re stack this allows you to move plates from the output stack to the input stack Full Screen Plate select this to see a full screen view of the counts tab plate Press Esc or right click the mouse to close this view Help menu Click this to get help on the use of the software At any point during operation pressing the F1 key will cause the appearance of help information relevant to that particular operation If your software malfunctions you can select Create Error Report to generate a report of the software status This will help the service person handling your case The report is created as a zip file on your compute
9. v2 0 21 22 23 26 Number of flashes e 5 1 5000 Size of the scanned area eeecose 4 5 1 4 5mm eeeeee 0 85 Z value target Z high sample zZ low sample a g c E a g 1 H Tl R 5 H r Height sample TRF high sample TRF low sample a0 oo O bos lt gt 384 OptiPlate e Homogeneous TRF Laser lt Back Next gt J Cancel If Plate dimension optimization is selected you must have samples at least in the corners of the plate e g an assay plate with samples in every well would be suitable The sample for height optimization can be the same as the A1 corner well for plate optimization The sample for Gain optimization is a separate sample in well B1 Z optimization requires high and low samples in the positions shown in the screenshot 50 Using the Assay Start Wizard lt Assay Start Wizard Plate preparation i Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Plate preparation TA e 4 22 Number of flashes 10 1 500 Sample is B c D E F ka Fs lt gt 96 General e Fluorescein lt Back L Next gt J Cancel If flatfield correction is select a platemap appears showing where samples must be placed Every well should have an identical sample in it so that the flatfield correction can be calculated When you have ensured that your
10. Cave fttn standards on each plate Carve fitting Lardards on each plate where Label Fhuoresceri1 darnel 1 veg of type Average of the samples where Label Calc L Curve fitting standards on each plate Sample All samples wath index Cale 3 Avg of tyne Average of the samples where Label Fiuerescein t channel L Sampie All samples wath index Cn ee Oe Es ie oe i 2 view 5000250000900 a GQ f Color scale I Loprthmi hio 9900090090920099 mi O8 8 O0eO0 000707000029 All the calculations selected for the protocol are listed Note If a calculation error has appeared in Notifications the calculation that has been defined might disappear from the list with no results shown To see the results of these calculations click on them one by one in the navigation tree Each time you select a calculation the main pane in the window will show the results of the calculation 83 Result viewing B Assay 1 Results 5 x Elle Gdr View Tools Actions Hep o0 Se Forward Up Export Cak assw ID 1 Sriated EE AS AS EED EA S S ES A E ENS e SS ee eee 3 Avgo type Normally the Counts view will be open but if the calculation is curve fitting the Graph view will open automatically to show the curve This shows the points from the standards that were measured and the curve fitted to them Clicking the Show fitting info check box causes
11. Next gt Cancel Crosstalk correction optimization AlphaScreen Assay Start Wizard Advanced mode checkbox Click Next to continue Optimization info f Please select optimization s you want to run For advanced options check the Optimization info Optimizations to run Plate Dimension Optimized 13 11 2007 13 11 04 Crosstalk Correction Optimized 13 11 2007 13 11 04 Flatfield Correction Not optimized The plate map shows the correct arrangement of samples 54 Using the Assay Start Wizard Assay Start Wizard Plate preparation Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Plate preparation t 2 a s sl el7 s s ifu 12 13 14 516 17 18 18 20 21 z z z 3 AlphaScreen sample e e L e L gt AAE JHE n 1 on k e gt 384 OptiPlate e HTS AlphaScreen 384 OptiPlate lt Back Next gt Cancel Note The platemap for 1536 AlphaScreen is set so that samples can be transferred from 384 well plates shown in the map by a robotic system using 384 needles tips 8 Optimization progress s6 ceneri Fluorescein This is what you see if the optimizations are selected 55 Using the Assay Start Wizard until the wizard Finishes the optimization To cancel the optimization click Optimizing Flatficld SS Wallac
12. Test Plate 4 Fluorescein This is what you see if flatfield correction is selected 9 Results If TRF optimization is selected you will first be shown a window with a proposed delay Assay Start Wizard TRF results Please check the delay value and correct them if necessary Click Next to continue Delay results E 20 30 40 RK Delay 40 lt gt 384 OptiPlate e Homogeneous TRF Laser 56 Using the Assay Start Wizard You can accept this or move the cursor to the delay you want Then click Next Assay Start Wizard TRF results Please check the window value and correct them if necessary Click Next to continue Window results 200 300 Window a 1 b 1 Oa 1 b 0 5 lt gt 384 OptiPlate J Homogeneous TRF Laser Oa 0 5 b 1 a Window 200 a 5 Smax b 5 B S B max The proposed window setting will appear Once again you can approve it or modify it by moving the cursor Then click Next Note These windows will not appear if TRF is not selected The next window only appears if Z is selected 57 Using the Assay Start Wizard lt Assay Start Wizard z results Please check the flashes and modify it if necessary Click Next to continue 2 results 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 Flashes Flashes 1 z 011 lt gt 96 General J Fluorescein You will be shown the results of the
13. Z optimization and asked to check it and if necessary change the number of flashes to be used 58 Using the Assay Start Wizard Assay Start Wizard 2 results Please check the flashes and modify it if necessary Click Next to continue Results Raw results z 1 0 0 9 0 8 0 7 2 results 05 0 4 0 3 0 2 01 ttt 40 60 80 100 120 140 160 180 200 220 240 260 280 300 Flashes Flashes 74 z 0 72 gt 96 General e Fluorescein You can also see the results numerically Assay Start Wizard 2 results Please check the flashes and modify it if necessary Click Next to continue Counts 124658 141520 250035 180192 196078 z results 148347 181914 127346 241495 258211 164960 299633 25191 31178 When you have done this click Next The last window of the wizard will appear 59 Using the Assay Start Wizard 10 Wizard completed Assay Start Wizard Wizard completed Assay start wizard has been finished successfully If Start the assay is selected click Finish to start the assay IF you dont want to start the assay immediately uncheck Start the assay In cases where an optimization plate must be used Start the assay will be unchecked Click Finish to end the optimization Load an assay plate and re run the Assay start wizard Assay notes shown in Result Notification 2 selected Start the assay The St
14. additional information to appear beside the curve Curve fitting calculations Navision Tree z T 1 Curve fitting standards on cach plate Aan 10 7 Enine j Fernan om am md J Giras Cak 1 Carve Mitro standards on each piata Curve Itno standards on aach piate where Label Fluwescer darnel 1 535 W nen i i Carve tetng star gate 2 meget oe vew OSametes on plate Chew terre ete 84 Result viewing If you click on one of the unknowns listed in the right hand pane the intercept with the curve from the response of the unknown and the appropriate concentration will be shown on the graph If the response of the unknown is outside that of the range of the calibration curve no intercepts will be shown on the graph The response and concentration values will be listed in red instead of the normal black To edit the curve select the Cale button from the toolbar A new window appears showing a curve and various curve fitting parameters see Advanced parameters as well as the concentrations of the standards and the units Assay 7 Standard curve fitting A Focescentt Label Caleulstons ve fitting standards cn anc TUOrescersl harnet 1 535 d type feting Leg Advanced Parameters Avg of ty 3 Avg of type None None dick Advanced Parameters Edt Mtting Parameters There is a button Advanced parameters giving you the following options 85
15. front are valid labels i e all the necessary modules and components are installed Configuring your user interface The View menu allows you to select whether you want the Shortcuts bar and or the Navigation Tree or neither to appear Introduction to the UserInterface Navigation Tree Wallac EnVision Manager 1 11 File Edit View Tools Actions Help Q 0 8 4 Back Log C Navigation Tree x Ae Reader Control Protocols Results Inventory Dispenser Control Temperature control 3 Measurement Technologies SS Plates a Samples i Barcode Settings M Reader Settings gt Audit Trail lt Recycle Bin This gives you access to all of the functions of the workstation There are various folders e g Reader Control Results etc most of which have sub folders If you are in a sub folder you can move to the next higher level by clicking the Up button Click the Back button to go to the previous location in the tree If you have gone back you can go forward by clicking the Forward button Clicking the arrows beside one of these buttons allows you to see the possible locations in the tree and to jump directly to the one you want Clicking the right mouse button leads to a menu 14 Introduction to the User Interface Open opens the selected item New shortcut makes a short cut of the selected item Hide Navigation Tree closes the navigation tree Select it from the View menu to re
16. left corner furthest from you i e the side that enters the instrument first Note If you are using strip plates make sure the strips are not sticking up at all but are properly in position otherwise the plate may jam in the instrument Results will not be correct if strips are not properly in position If you are using barcodes these can be attached to the plate on the front or back long side or right short side depending on which is selected in Reader Settings 39 Options for running EnVision Note Correct plate labeling sample preparation and sample order on a plate are critical factors in obtaining correct results 40 Using the Assay Start Wizard Using the Assay Start Wizard To get optimum results with EnVision you should optimize the measurement technology and plate combination you are going to use in your measurements Assay Start Wizard The Assay Start Wizard guides you through the process of optimization If you have an optimized plate and technology combination you can still use the Assay Start Wizard to start your run Use of an optimization plate For most technologies you do not need a special optimization plate although you can use one A normal full assay plate is sufficient When you have done an optimization with an assay plate you can start to run it directly from the Assay Start Wizard Note Special luminescence and AlphaScreen require an optimization plate to be used Click the
17. lt gt 96 General Fluorescein lt Back Next gt Cancel The plate map parameters are as follows 101 Advanced optimization Number of flashes You can select the number of flashes to be used This can be different from the number set in the label parameters E g if in the label parameters the number of flashes is set to 30 you can change it to e g 10 to speed up the optimization process With AlphaScreen the measurement time is fixed and it cannot be changed The measurement is made with an AlphaScreen sample Size of scanned area Enter the size of the edges of the area of the wells to be scanned See the table for typical size settings Size settings No of wells Spacing of Typical size Allowed centers setting range mm mm mm 1536 2 25 1 8 1 4 5 384 4 5 27 1 9 0 96 9 6 3 1 18 Note There are 10 movements in the scan in both the horizontal and vertical directions producing a measurement of 121 points in an 11 x 11 array 102 Advanced optimization Z value target Give the Z target value This is typically between 0 6 and 1 0 When you click Next the optimization will begin If Plate optimization has been selected a window will appear with four tabs each with a picture showing the distribution of signal intensity across a corner well These pictures are to enable you to mark the center of each well This will be the area with the most inten
18. or Small icons in a group depending on what appearance you like All these operations are possible with the menu under the right mouse button Note The Advanced Use group contains all the main icons from the Navigation Tree Introduction to the User Interface Recycle Bin The Recycle Bin folder stores deleted results protocols plates etc Wallac EnVision Manager 1 11 Simulation mode File Edit Yiew Tools Actions Help E B P iD ol Back Log Out Start Latest Restore Empty Navigation Tree x Ae Reader Control i Protocols Name Deleted by Delete Date 9 Results 100100 New Protocol aaa 20 11 2007 15 28 46 amp 4 Inventory Dispenser Control Temperature control To restore a deleted item select it and click the Restore button on the toolbar Alternatively you can click the right mouse button A Restore button will appear Click this The item will disappear from the Recycle Bin and will be restored to the location from which it had been deleted Note When a protocol is deleted all results measured with that protocol are also deleted When you restore a deleted protocol any results that were deleted with it are not restored You can also permanently delete all items from the Recycle Bin Click the Empty button on the toolbar Confirm that you want to permanently delete all items by clicking OK or click Cancel to avoid deleting the contents of the Recycle Bin Introductio
19. the syringe to the empty postion Ladis expelled ite the baud reservar OFM syringe Drive the syringe to the full position Uguid s aspirated from the kqsd reservow Cre Click Next when you have selected this option Select the pump or pumps with the syringe s you want to empty See Show advanced options for some extra parameters you can edit Click Next 127 Dispenser control Wallac EnVision Manager 1 11 Simulation mode Ble Edt View Toots atos tep 0 6 7 Q tack tog nt emote sd Dispenser Control 2 proved Click Start or press PUMP button on the dispenser to Empty syringe Cae Com Click Start or press the PUMP button on the dispenser The syringe will be emptied into the liquid reservoir Temperature control activated Current 22 53 C Ready Emptying syringe You can repeat this operation if required to empty the syringe of PUMP2 Click Finish The Maintenance display will reappear 128 Dispenser Control Fill syringe This operation drives the syringe to the full position Liquid will be aspirated from the liquid reservoir to fill the syringe Wallac EnVision Manager 1 11 Simulation mode Temperature control activated Cumont 22 63 C Ready Rinse Dispense a selected volume of baud into the waste container Ont tubings Apae wazh kasd to fil the tubing Amal excess 6 dhiperiod into the waite container ORetrieve liqu
20. typa Average of the samgies where Label Fuorescming s channi 535 Sampin All samckes with andere 1 Curve fiting star OCak 4 Peak ol type Maximan vakas of the someles where Label Floresen channel t 535 Sampie AF samehes with inden 2 Arg type aade 4 Peakot type 1 2 2 4 s g 9 10 u 12 vew 42 gt 4 4 amp 4A 4 4A Sy 4B By BM Oco Note The raw data is not changed by adding or removing calculations 96 Result viewing To delete a calculation click the Calc button to get the calculation window Select the calculation you want to delete then click the Delete button then click Apply Assay 7 Standard curve fitting YX FLX Apply Cancel Calc 41 Delete Protocol General settings Plate 1 Wells selection Group 1 Measurement Fluorescein 1 Label Calculation O Calculations 1 Curve fitting standards on eac Average of the samples Fluorescein 1 Channel 1 535 2 Avg of type Sample type po Ava of type All with Index If you click the Cancel button instead of Apply the calculation window will close without making changes 97 Result viewing Export Export Export To File O Export To Printer File name DefaultDataFolder gt lt Date gt lt ProtocolName gt _ lt AssayID gt cs Export format Plate 2 wth well headers Assay information to include Plates Basic assay information plates N
21. unit S10 Coreeetraten 1 5 2 w gt Rens wd be re calodsted when you press the Apply button 91 Result viewing Show gridlines this toggles the gridlines on and off STD Concentration J 2 s00 320 000 Slope at 50 binding 300 000 ia ii 421 1 280 00 260 000 Intercept 240 000 64810 220 000 D20 200 000 23 13 180 000 EA 160 000 173 0 140 000 ED 80 120 000 369 2 100 000 Variance Ratio 80 000 0 000 100 200 300 400 500 Calc unknowns type in a response value and see the concentration calculated from the curve The response value must be within the range of the curve ect outliers click Advanced Parameters Edit Fitting Parameters w centration unit gt Concentration 5 500 320 000 Inspect Unknowns Tt Slope at 50 binding aoaoga Response 66914 j L 421 1 280 001 Concentration Ss 260 000 E Intercept 220 000 D20 200 aia 23 13 180 000 if ED 50 160 000 173 0 140 000 t ED 80 120 000 369 2 100 000 ji Variance Ratio 80 000 0 000 100 200 300 400 500 92 Result viewing Other types of calculations If you select a calculation that does not use a curve e g Average of type then the display is different 1 2 3 5 7 s 10 n nR CELILILIILILII iaa EBL LE amp amp amp amp A BB oo If you now click the Calc button the display shows details of the calculation
22. you get the greatest range of values for each curve but you cannot compare the size of curves with each other directly Show points select this if you want the actual points to be visible It requires adequate magnification to see the points 24 Introduction to the User Interface Reader control window Temperature Wallac EnVision Manager 1 11 Simulation mode Fle Edt View Tools Actions Help 2 e 3 28 OOO Log Ot Stet Latest Ese ws Reader Control Temowratire control xtvatod Curent 22 35 Roxy YA Load Rosta iS Idle Select protocol vaki protocols marked wrth and dkk Ruri button to start the assay Protocol Qordard orm fiting Label A Source Charnel 1 MO Assay 004 Plate 001 repest OL Counts Geach Temperature Events Temperature GD Assay examples CQ Wala protocols Q Reags H AG inventory T Dispensar Control Temperature corral Measuremert Technologes UG Reader Settings Be muse trast E Recycle Bn Humidity a Man arn aA Temperature and humidity values are shown in this chart These values are the temperature inside and outside the instrument and the humidity inside the instrument Consistent AlphaScreen measurements require a stable temperature This is ensured by the cold plate which is in close proximity to the assay plate You can check from this chart that the temperature has remained stable during the time of your measurements See Temper
23. 2104 9010 02 March 2008 User manual EnVision Software version 1 12 Wallac nVision Manager 1 11 Simulation mode lite an Reader Control Temperature conte actwated Current 22 35 C Ready Ae Idle Select protocol vaks protocols marhad with are cick Baa button to start the ancy Protecel AES 405 Labet Absorbance 45 Source Absorbance value MO Assay 003 Plate 001 repeat OL CEREN Temperature sel ne A A E CIIL 1002000000000 E Trak omer wet O06 0606666666 _ amp amp COS 2S OO 6246 6666666 O2622266660 2060006028662 2006660680606 pP PerkinElmer precisely 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 Pai G EC GN OB a cisiecesensdecdvcseonanaweidenahonzaceniesccindt seroso steedde ossee eitast 3 SOLE Wale Stall AWW ss aca a hades s a entu ga sda ii aaa 5 StA UDan e e i Neel E R a E E E i ie Sah 5 Shut GOWN essun sin A aiaa E E asa RSS 6 Using thismantal aei E E E A e 7 Conventions sediar a a E N E A coments 8 Introduction to the User InterfaCe seesessossesossossesessoesessossesossossesossosseseoss 9 Reader control Window sissyeves ccveavacladosdvacastesacasengesecd yeddnanstoeapeiastaontaeasceadantaves 9 Reader Control toolbar is ieseis a casssecievsedsdissead es Lossensanueesesourdeaees teaseatede
24. Assay 7 Standard curve fitting Y X X Apply Cancel Calc41 Delete E S Protocol General settings Average of the samples E Wells selection Group 1 D Measurement Fluorescein 1 Label Calculation Calculations p 1 Curve fitting standards on eac Cale 1 Curve fitting standards on each plate 2 Ava of type Sample type 3 Avg of type All With Index 93 Result viewing Adding additional calculations Note If a calculation was not added to the protocol prior to measurement it can be done afterward First click the Cale button in the Results window B Assay 7 Results File Edit Yiew Tools Actions Help 9 0 2 S f Back Forward Up _ Export Calc r E B Assay Calculations por coms am z en aad 1 Cale 1 Curve fitting standards on each Fluorescein 1 f O Calc 2 Avg of type Average of the sa e a f O Calc 3 Avg of type Average of the sa 1 Curve fitting star 2 Avg of type fr 3 Avg of type 94 Result viewing In the calculation window that appears select the arrow beside the Calc button to see the list of calculations Assay 7 Standard curve fitting V X FIX Cancel Cale 41 E Protocol Gi Es Plate 1 wel 3 Label ratio l 4 Label addition a 5 Label subtraction f 20 Peak of type 21 Sum of type 22 Avg of type 23 C of type 24 Standard deviation of type 25 Copy 27
25. Dispenser Control An animation will show what is happening ode b 7 ot t Wot i Dispenser Control Targato contral activated Catwnt 22 46 Gundy You can repeat this operation if required to fill tubing for PUMP2 Click Finish The Maintenance display will reappear Retrieve liquid allac EnVision Manager 1 11 Simulation mode Ble Edt Wew Tools actons tip 0 8 7 Wa Formas logon rman rea 5 A Cont Dispenser Control 4 1 IB Protocols Markenance settings Lovertony Dispenser Control Inttiaieation Vemperense conte Dauno 1 irtsakzod Damo 2 irtsaknad 4 a gt Serghes m aod tna Oninse Rander sattes Cupense a selected voise ot baud ino the waste container Aud Trak Recycle Bn On tubiegs hapaa rec bead to fl the tubs A ermal exces chupmenind into the mnte contara Retrieve liquid back to the bottle Return baud from the syringe and tubing to the kad reservor O Test dispense Dispense a set vohane to the maste container Ottmety syringe Orve the orga to the empty pouten Ligad mapeled into the kyai reservor OPA syringe Crave the syringe to the ful postion Liquid is aspirated from the kgad reservor Next gt 121 Dispenser control Returns liquid from the syringe and tubing to the liquid reservoir use this if the liquid is expensive reagent Click Next when you have selected this option Wallac EnVision Manage
26. Flatfield 28 General 31 Curve fitting standards on each plate 32 Curve fitting standards on first plate only 33 Curve fitting blank corrected standards on each plate 34 Curve fitting blank corrected standards on first plate only 35 Validation Note The calculations available depend on the measured labels and the measurement operations Select the calculation type you want to add The new calculation will be added to the calculation list 95 Result viewing Assay 7 Standard curve fitting YV XIP X Apply Cancel Calc20 Delete Protocol General settings f A Eas Plate Maximum value of the samples Wells selection Group 1 ED Measurement Fluorescein 1 Label Calculation O Calculations I 1 Curve fitting standards on eac Fluorescein 1 Channel 1 535 2 Avg of type Sample type 3 Avg of type 4 Peak of type AlI With Index There may be parameters you need to set See Calculations in the Reference manual for more information Click Apply to obtain results using the calculation The Results window will open with the additional calculation added Cak 1 Curve feing tandede on nach plate Curve fitting standards on mach plate where Label Phaoresceindt channel 1 S35 Oars Preses 1 O Cak 2 avg of type Average of the semeles where Label Cak 1 Curve fitting andards on each piata Sampie All sampi wth ates s l O Cak 3 Avg o
27. Latest P fF X Filter Open Delete Results Fiter applied T Protocol I Amount Sho 5 0 resi F Time From rero oo Te esjorjeoor fisioo 1s Ne 7 a 3 9 23 01 9 Standard curve fitting 23 23 LUM Single 23 01 21 59 00 23 01 20 59 08 TRF Eu Top Single 23 01 2007 14 57 52 23 01 2007 14 58 01 LANCE EuJAPC Top Dual 23 01 2007 14 57 32 23 01 2007 14 57 45 FP FITC Dual 23 01 2007 14 56 58 23 01 2007 14 57 08 FP FITC Dual 23 01 2007 14 56 46 23 01 2007 14 56 56 FI FITC On the Fly 23 01 2007 14 56 28 23 01 2007 14 56 33 In the view window a number of selection criteria appear If you have many protocols use a selection criterion to limit what appears so that you can more easily find the one you want More than one selection can be used at the same time Note All results of a protocol can be viewed under the protocol in the Result folder 75 Result viewing Results filters The selections are Selection by protocol Select the protocol from the drop down list Selection by amount You can select how many of the most recent results are listed Selection by date and time Give the first date and time and the last date and time for the period you want to select Note Each time you make a change to the selection settings you must click the Refresh button Any assays in the selected range will be listed in a pane below the filter options along with their date and time of r
28. Start button to start the assay start wizard 41 Using the Assay Start Wizard Steps in running the Assay Start Wizard 1 Welcome to the Assay Start Wizard Assay Start Wizard Welcome to the Assay Start Wizard This wizard will help you to start the assay and run the required optimizations if necessary Also using this wizard it is possible to perform various optimizations without starting the assay If you want to perform optimization s please Follow instructions in the wizard IF you just want to start the assay please load the plate with samples into instrument To continue click Next TE Click Next 2 Select action Three option buttons allow you to select your action Select action Select action A Please select action want to proceed Click Next to continue Optimize existing protocol O Run existing protocol O Create new protocol Optimize existing protocol selected This operation will let you choose the protocol and optimize run it Select the option button and click Next 42 Using the Assay Start Wizard The table shows the different steps of the wizard depending on the option you have selected Create new Optimize Run existing protocol existing protocol protocol Select folder for Select protocol new protocol Select plate Select technology Select optimization Define plate layout Run optimization Finish The following explanation
29. To tbe vote 0 2000 yt m 70 EEI Aspiration tube voine h Total tube voma lt a Select the pump or pumps with which you want to do a test dispense Select the tip mount to be used This must be the one in the instrument as shown by the mark See Show advanced options for some extra parameters you can edit Click Next Temperature corso activated Current 22 64 C oady Click Start or press PUMP button on the dispenser to Test dispense 125 Dispenser control Click Start or press the PUMP button on the dispenser Liquid will be dispensed to the waste container An animation will show what is happening Wallac EnVision Manager 1 11 Simulation mode You can repeat this operation if required to dispense liquid from PUMP2 Click Finish The Maintenance display will reappear Empty syringe This operation drives the syringe to the empty position Liquid will be expelled into the liquid reservoir 126 Dispenser Control Temperature control activated Curent 22 64 C Ready E iode settings Orme Raso Settings Dispense a selected vohane of kaud into the raste container Aydt Trad O Recyde tn O Mi tubings Aspa sta arca kgad to Fi tha hbng A amal ascos x departas into the waste contaras O Retrieve Bquid back to the bottle Return kepad from the syringa and biting to the baad reservar O Test dispense Dripnee a set vohama to the wane cortaner Ompty syringe Drive
30. added to the list under the Overlaid curves button 22 Introduction to the User Interface aly NNW NA WN ie Ae Ad Va ad WY a A NY tt ply MN a Ms YW YVR AP WY i vin Fup jah Ar vig LAAN ya N a A gf Ny ae WAM Wa a al AY MYT VA ey MAA YA ANA A VP A AY A View Plate size to Fit Overlaid curves Qaat aoz dd curve Remove curve Remove all Curve scale IV Relative to greatest IT Show points When you have selected all the curves you want click the Overlaid curves button The selected curves will be displayed with measured values plotted against time 4 0 0 0 1 0 2 0 3 0 4 0 5 Time sec View C Plate size to Fit 7 Overlaid curves Oa aoz Add curve Remove curve Remove all 23 Introduction to the UserInterface To remove a curve from the display select it from the Overlaid curves list and click Remove curve Click Remove all if you want to get rid of all curves from the Overlaid curves list Curve scale These check boxes are only visible if the Plate option is selected Relative to greatest all the curves are scaled relative to the greatest value of all the curves on the plate This enables you to compare all of the curves but it may hide the variation in values for each curve if there is a large difference between curves If this check box is not selected then each curve is scaled independently This way
31. ai ced as haven Ea RSi ase 83 Adding additional calculations eeeeeeeeseeeeesesesesressersresrersessrerrresreseese 94 EXPOr sate honeran R dest ah A A AE RAITA 98 Advanced Optimization e sssesssocesooesoocessecesoccssocesocesoosssoesssccssocesocessosesse 101 Plate dimension optimization ssseeseseseesesseesreerersersrerressteseresresseseresreses 101 Measurement height optimization sseeseeseseseesesserssesreeseesresrresseseresresse 105 Detector gain optimization FI and FP ee eee eeseceseceseeeeeeeeseeenaeenes 107 Crosstalk correction opt Enh lum and Ultra Sensitive lum 110 Crosstalk correction opt AlphaScreen and HTS AlphaScreen 111 Contents Dispenser COmtroll siccvesssssvccesesvd cosvsenvscenesovedoszscnceeugvessesessvencesvsvensssavevarssnvers 115 IVa ING ead wc aes at Coe E E Saath ane aes teeth te ee Aca 115 Tri Gai ZattOn on sssini saree eet eeee ibi aiia 115 RNS Cs cise cele Sa ee ie eee eed a aa 116 FI GUO Sahel cache bce evap Re dada hte a aE Se 119 Retrieve liGuid 32 02 si ssn gaiesiitutitee nd nantiedwn iia 121 Test dispense seis g etcetera sh tone eae east EE 124 EMpty SYN SE ent ot ee ete eet alata BNE ESAE 126 Pull Syringe nasin easa eel Chere aeeds 129 SHOW advanced Options cessit soie ecese riesa eiiie iei 131 POC MTS na e a a Cue Nh Rta tale a Onan E Ci R a 133 Dispenser d idelineS nnani ensi fovea A a Ea acta eae Le eae 135 When starting operation aies
32. alk results To view the raw results select the Raw results tab Click Next to continue Raw results Crosstalk results Glow Correction Factor 100 00 You can also click the tab to see the Raw results Assay Start Wizard Crosstalk results Please check the calculated crosstalk results To view the raw results select the Raw results tab Click Next to continue Crosstalk results Time ms Counts 0 52688 66 96005 85703 20943 27403 60964 27999 99123 27905 43567 40131 Click Next to go to the final window of the wizard 110 Advanced optimization Crosstalk correction optimization AlphaScreen and HTS AlphaScreen After you have run the optimization as described in Assay Start Wizard the results will appear Assay Start Wizard Crosstalk results Please check the calculated crosstalk results To view the raw results select the Raw results tab Click Next to continue Raw results Crosstalk results After Glow Correction Factor E Time ms 80 380 690 990 1290 1592 1890 2188 CT 0 00 0 00 830 10 0 00 656 34 0 00 622 66 246 25 Glow Correction Factor 1281 05 Yo Bleach Correction Factor Bleaches CT Crosstalk results Please check the calculated crosstalk results To view the raw results select the Raw results tab Click Next to continue Crosstalk results Counts 458982 499239 499153 475713 477052 475166 463860 455383 462853 454743
33. ance from the detector Identical samples need to be used in each well of the plate The system determines the differences between each part of the plate The correction factors calculated are applied to subsequent plate measurements Since the position of samples for this correction cannot be combined with the positions for other optimizations this correction has to be performed separately Crosstalk correction optimization special luminescence This allows the glow crosstalk to be measured and corrected for A single normal sample and a blank are used for the glow crosstalk measurement Additional blanks are used to obtain an average blank value See Special luminescence for more information about this optimization An optimization plate must be used Crosstalk correction optimization AlphaScreen This allows three types of crosstalk to be measured and corrected for These are glow afterglow and bleaching In order to measure these different types of crosstalk several configurations of samples are needed on the plate The wizard shows what is necessary See AlphaScreen for more information about this optimization An optimization plate with a specified plate map must be used 38 Options for running EnVision Plate orientation The figure shows how a plate is to be orientated when loading it manually into the instrument The same also applies when loading the plate to a magazine of the stacker The A1 position must be in the
34. anced options if you want to see or change certain basic parameters If you do not select this option then the default values will be used There are two sets of identical parameters for Pump and Pump2 131 Dispenser control 11 Simulation mode DER elp P D ogOut Start Latest gl Di Control al Dispenser Coni Temperature control activated Current 23 15 C Target 22 00 C Adjusting e Settings Rinse Pump s F Pump 1 miPume 2 Tipmount 5 Real Time Pre96 Dual Tip V Show advanced options Pump1 Pump2 Total volume 500 10000 pl fzooo Total volume 500 10000 pl 2000 Speed 100 500 p s ko Speed 100 500 pls 200 Tip tube volume 50 2000 pl 200 Tip tube volume 50 2000 pl 200 Aspiration tube volume pl 270 Aspiration tube volume pl 270 Total tube volume pl 470 Total tube volume l 470 Remember settings Total Volume This parameter determines the volume of liquid aspirated For Rinse the default is 2000 ul and the range 500 10000 ul The parameter name is Total volume because several dispensings occur and this parameter is the total volume used for all the dispensings For Fill the default is 50 ul and the range 50 10000 ul The parameter name is Volume For Test the default is 50 ul and the range 2 475 ul The parameter name is Volume 132 Dispenser Control Speed 100 500 ul s This para
35. art Wizard e Or press the START button on the instrument These options are described as well as the use of a stacker with barcodes or without barcodes Note If you later change the label or plate parameters used in optimization you must redo the optimization Options for running EnVision Types of optimization Plate dimension optimization All technologies This enables EnVision to determine the exact positioning of the plate being used The position of each of the four corner wells of the plate is measured It requires a plate with samples in each of the four corner wells using the label you intend for normal measurements E g a full plate is suitable Measurement height optimization All technologies This enables EnVision to determine the optimum focal point for measurement A single sample is required for this Unless you choose otherwise the strongest sample will be identified and used The plate is measured over a range of 35 Options for running EnVision vertical positions and the one giving the highest signal is determined Monochromator wavelength optimization Abs This optimization determines the wavelength at which maximum absorbance occurs A single sample is used Select the lower and upper limits of the wavelength range Select also the step length between wavelength measurements and the number of flashes A plot of wavelength against counts is displayed and the wavelength for the lowest coun
36. art the Assay check box in the last window of the wizard allows you to select if you actually want to run a plate using the optimization just done or if you just want to save the optimization information For optimizations where a special optimization plate is not necessary this box will be checked by default If an optimization plate is necessary the box will be unchecked by default In the latter case the window will just close when you click Finish If Start the assay is checked the currently loaded plate will be run and the results displayed You can add a note that will be included in Result Notification 60 Running an assay without the Assay Start Wizard Running an assay without the Assay Start Wizard There are different ways of operating EnVision depending on the combination of plate loading method and barcode setting you have Read the text describing the combination you are going to use Note Before beginning any loading make sure your plates are orientated properly Running an assay no protocol barcodes no stacker 1 If the plate carrier is in the instrument press the LOAD button or click the Unload button on the toolbar The plate carrier will come out 61 Running an assay without the Assay Start Wizard Note The software button shows Unload if the plate carrier is in the instrument and Load if it is out 2 Load your first plate Make sure the plate is orientated correctly with the A1 po
37. ating Select this check box and then select the temperature for the liquid in the liquid reservoir The range is 25 to 60 C Advanced settings Select this to see extra parameters for both Pump and Pump Connected to tip select which tip you want each pump to be connected to 1 2 or None Aspiration speed is x of dispensing speed the range is 5 to 100 and the default 70 Aspiration tube volume set the volume for the aspiration tube if it is different from the default 270 ul The allowed range is 50 to 700 ul This value is used in Show 134 Dispenser Control Advanced options for the various dispense operations to calculate the Total tube volume by adding it to the Tip tube volume Dispenser guidelines When starting operation Perform Initialization when you have switched on EnVision software is rebooted after being completely shut down you have pressed a PUMP button during dispenser operation The software will tell you that you cannot run assays requiring dispensing if you have not initialized the system Before daily work At the beginning of a working day we recommend that you perform the following operations depending on the situation if the tubing has been rinsed and emptied when operation finished the previous time recommended procedure then Fill the tubing with reagent if reagent has been left in the tubing not recommended some may have been lost through evaporation
38. ations you want to be done or redone Check boxes allow you to make this selection If you click Next without selecting an optimization the wizard will jump to the last window Note Check Advanced only if you want to inspect or change the selections made by the software This example continues without Advanced being selected See Advanced Optimization for examples of screenshots that appear when Advanced is selected Click Next 48 Using the Assay Start Wizard 7 Plate preparation Assay Start Wizard Plate preparation Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Plate preparation e ol0 44 42 Number of flashes o 10 1 5000 Size of the scanned area 9 1 9 mm 0 85 2 value target 6 Z high sample Z low sample Height sample Gain sample a0 bn The first example shows where samples must be placed for the optimizations selected when Detector Gain is one of the optimizations The second example shows where samples must be placed for the optimizations selected when TRF is one of the optimizations 49 Using the Assay Start Wizard Assay Start Wizard Plate preparation Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Plate preparation ila a e s s 7 8 s 01 12 3 16 5 16 1 i
39. ature control in the Reference manual for how to set the temperature for AlphaScreen 25 Introduction to the UserInterface The temperature control system allows you to raise and stabilize the temperature inside the instrument for cell based and enzyme measurements which require e g 37 C See Temperature control in the Reference manual for how to adjust the temperature control The side bar on the right of the window gives you the following options Values These are the current temperature around the instrument Ambient and in the instrument at the position of the plate Chamber The humidity in the instrument is also given Show raw data click this to see more detailed temperature information from the sensor inside the instrument at the position where air is sucked out of the instrument right back corner at the conveyor level This is a little above ambient the upper sensor above the plate the lower sensor below the plate conveyor The chamber temperature is obtained from the upper sensor unless the required temperature is set In this case the chamber temperature is the average of the upper and lower sensors Legend This identifies the different curves on the temperature and humidity graphs 26 Introduction to the User Interface Reader control window Events Wallac EnVision Manager 1 11 Simulation mode d im ratx example latest gt Idle Select protocol vabi protoc
40. caisucssdsdtegs aes Sgadsy tees asndacvosdadamierineneganssaueines 135 Before daily Work lt 1 sgrcsvesissiasazenisaaveceatie deseds woaaaaesouracrasvaseatesssegenaonsaats 135 After Operation 22 43 c ae GR eS ee ee 136 Changmg atip MOUN Gc BA etal Seale Bs teat areas ade 136 PANN C A OEE AA E EE S O 139 Trademarks Trademarks Wallac LANCE FP and EnVision are trademarks and PerkinElmer AlphaScreen and DELFIA are registered trademarks of PerkinElmer Inc Windows Windows XP and Windows Vista are registered trademarks of Microsoft Corp in the U S and other countries Chapter 1 Introduction Introduction Introduction The EnVision high throughput screening microplate reader from PerkinElmer Life and Analytical Sciences is a complete platform for quantitative detection of light emitting or light absorbing markers It is suitable for measurement of fast or glow luminescence absorbance fluorescence intensity fluorescence polarization FP high sensitivity time resolved fluorometry DELFIA and homogeneous time resolved fluorometry LANCE Very high sensitivity measurements can be made with the Ultra Sensitive luminescence option A laser equipped model allows AlphaScreen measurements to be made HTS AlphaScreen allows faster throughput than standard AlphaScreen Another option with an external laser enables enhanced time resolved fluorescence measurements The monochromator option allows a wide range
41. e prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Scan plate for strongest sample Plate preparation 1 2 3 4 s5 e6 7 8 9 40 11 42 Number of flashes 1o 5000 Gain sample Blank sample in Ole gt 96 General J FP Fluorescein Dual lt Back Next gt Cancel Note For AlphaScreen and special luminescence you must use an Optimization plate with the correct arrangement of samples to do crosstalk correction Crosstalk correction optimization special luminescence Assay Start Wizard Optimization info Please select optimization s you want to run For advanced options check the Advanced mode checkbox Click Next to continue Optimizations to run Plate sion Not optimized z Not optimized Crosstalk Correction Not optimized Flatfield Correction Not optimized The plate map shows the correct arrangement of samples 53 Using the Assay Start Wizard Assay Start Wizard Plate preparation f Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Piate pr epenen i i eB a z Measurement time sec sled ih 01 60sec E Blank sample i fa K al 0 o Crosstalk sample o SS 384General US LUM 384 cps lt Back
42. eeeeeeseeereesrrsrrsrresresree 42 Contents Running an assay without the Assay Start Wizard sccssscssssscesees 61 Running an assay no protocol barcodes no stacker eee eeeeeeeeeeeeeeeee 61 Running an assay with protocol barcodes no stacker eee eeeeeeeeeeeee 64 Running an assay with a stacker no protocol barcodes cesses 66 Running an assay with a stacker and protocol barcodes eee 67 Buttons on En Vision snssscneriarisnuanin siati anii 69 START al UL 6 e eee Enea Venere riers DRC MU nae terse tne Rery EE a ate errr 69 STOP DUON ante ete cilia aie tela di ele nten 70 LOAD DUttOn fuk hk oi eee riaa anessa eE an ia eae 70 Order of Dual label measurement in single detector instruments 70 BROS ULEE Viewing 5 2 ce dacs ca carecnucetyiaceicencosatncbesueseeqetievecntnseseesadecdedesiatuoretieskedeoies 75 Res lts fitefiS lt 2 ananin Sota tue cect ot aSa E ee A ato an 76 Results toolbar buttons tiacsco8 cecegeeic a hed aes cam eigen ieee eeeeniis 11 E E E E ate ee ic ati dpe hie eda alate oaanuaien 78 Results Window 225 cen eG Rh id GS a ee eee 78 PRS SAY acta E Sie ae E e E ha A E Ee ha eee ds 78 Protocol sii c3ectsass sateen ee ieee ied eed eee eS Wied Ei aS 79 Notifications sere en es tee ek alae cet ie cok tod a 80 Plate states ia eR eden ae elas Miata ke Road oles ti tated 80 GIT OU asics addi su cenedaxsananasgenaccashasesdesoeynncaavesaceusaeee E 81 Calc lati ons nshan aa a
43. erence manual for detailed information about all types of parameter setting Part 1 Introduction Tells you how the system works in general Part 2 Operation Takes you through use of the Assay Start Wizard including optimization and proceeding to running assays Part 3 Results Describes result viewing and output Part 4 Advanced Optimization Explains some of the additional steps involved in optimization if you choose to use the Advanced selection in the Assay Start Wizard Part 5 Dispenser Control Describes the software parameters used to control operation of the dispenser option Gives guidelines for daily use of the dispenser option Please refer to the Instrument manual for routine maintenance of the dispenser Tip Mount and Temperature Control settings are described in the Reference manual Introduction Conventions used Buttons or other software items to be clicked with the mouse are in bold text e g File Items in the main Navigation Tree are in bold italic e g Protocols Buttons on the instrument that need to be pressed are in block letters e g START Note The term AlphaScreen refers to both standard AlphaScreen and HTS AlphaScreen unless a distinction is specifically made The term special luminescence refers to Enhanced luminescence and Ultra Sensitive luminescence Note If there are features described in this manual which are not in your software it means that the feature is an option which is not
44. erep esse uara jeving en back yf the bottle peorse tn we a a A i You can repeat this operation if required to retrieve liquid from PUMP2 Click Finish The Maintenance display will reappear 123 Dispenser control Test dispense This operation allows you to dispense a set volume into the waste container Wallac EnVision Manager 1 11 Simulation mode Q Ompeenee Contd eetiokoticn Tengo cote Frp patat Measurement Tecrrelogies ek mre Llereep 2 tneisons Merce E barcode setteas Oime WG Reader Settnos Oncerse a selected vokane cf kasd nto the maste cortaner AR Trot D Recyde on Dll tubings Aspirate enough kaud to fil the tubing A small excess is dispensed into the maste contare O Retrieve liquid back to the bottle Reba kasd Irom the syringe and tubing to the laud rezervor Olen Capen a ont valine to the wate contar Otmoty syringe Orive the syringe to the empty position Liquid is expelled into the iqud reserwow Fil syringe Orive the syringe to the full position Lipad is arated freee the kgad reservar mE Click Next when you have selected this option Note When the instrument lid is open you can by hand or with a tool move the tip mount outside the instrument so that you can observe the flow of liquid through the tip In this case the tip mount used must be identified 124 Dispenser Control Vome 2475 1 w 20 Senet 100 500 1a Tete wine soat 200
45. has been too high so that the detector is saturated A red cross along with the Assay ID icon shows an error Other notifications are in blue Plate B Assay 7 Results o x Eile Edit View Tools Actions Help 9 90 39 5 Forward Up Export Plate 1 Assay ID 7 Simulated woe EP Assay PLACE TD ose eeeeeeeeeeeeeeeeeeeeeeeeceeseeeeee een eeereeseneees B Protocol a LD Notifications z cart 23 01 2007 15 0 Fluorescein 1 emperature at start chamber ons 809 Be emperature at end chamber sine 2009 C S Calculations emperature at start ambient Beer mgt 1 Curve fitting star Fremperature at end ambient cue 22 C 2 Avg of type fim Giby ab E E E cas gis sins cdee dees ves hcaaceaassseetene stains 20 9 fi 3 Avg of type fim di ty sac ends E s see E AEN 21 Selecting this will show information about the plate 80 Group Assay 7 Results Eile Edit View Tools Actions Help Result viewing loj x Q 0 8 Back Forward Up A Export E E Assay B Protocol AD Notifications B s Plate 1 ao Fluorescein 1 EH Calculations fe 1 Curve fitting star 2 Avg of type Raw results for channel 1 RFU label Fluorescein 1 3 Avg of type i e 8 i 0 een fee 1D 7 Simulated View Color C Value Color and value size to Fit x Repeat 1 Color scale T Logarithmic
46. id back to the bottle Ratuzn kaad from the syringe and tubs to the byad reservoir O Test dispense Dispense a set volane to the maste container Otmety syringe Drive the syringe to the empty postion Liquid is expelled into the iquid reservoir OFA syringe Crave the syringe to the full postion Lid is asprated from the kgad reservoir Cra Click Next when you have selected this option Wallac EnVvision Manager 1 11 Simulation mode Temperahre control activated Curent 22 69 C Ready 129 Dispenser control Select the pump or pumps with the syringe s you want to fill See Show advanced options for some extra parameters you can edit Click Next Wallac En ision Manager 1 11 Simulation mode lo Cdt Yew Tools Actions tep 0 6 7 Bak tego i Let i erg Mareenance Settings WG rwertory Temperabare control Neenserart Tecioges Brees gt barcode sers t personae Click Start or press PUMP button on the dispenser to D Rocyde Dn Fill syringe Cae Cre Click Start or press the PUMP button on the dispenser The syringe will be filled from the liquid reservoir 130 Dispenser Control Wallac EnVision Manager 1 11 Simulation mode Fle Edt Yew Tools Actions Hep Filling syringe You can repeat this operation if required to fill the syringe of PUMP2 Click Finish The Maintenance display will reappear Show advanced options Select Show adv
47. id protocols are marked with Please select the valid protocol before continuing Click this balloon for more info voeococeee Iser 9 44 Using the Assay Start Wizard 4 Select plate Assay Start Wizard Select plate ore Please select the plate for the new protocol and click Next to continue Select plate Name Rows WelllX WelllY well4x Welly 481536 AlphaPlate NEW 32 48 11 03 7 88 116 78 77 63 81536 CulturPlate NEW 32 48 11 03 7 88 116 78 77 63 1536 General 32 48 11 01 7 87 116 76 77 62 381536 Greiner 32 48 10 94 7 74 116 6 77 44 91536 Greiner Low Base 32 48 11 11 7 91 116 65 77 49 3H 1536 OptiPlate NEW 32 48 11 03 7 88 116 78 77 63 Select the plate to be used The name of the plate selected will appear at the bottom of the window Click Next 5 Select technology Assay Start Wizard Select label Please select the label for the new protocol and click Next to continue Select label Absorbance HTS AlphaScreen Fluorescence Intensity O Fluorescence Polarization 5 US Luminescence Luminescence DELFIA Time resolved Fluorescence Wallac TRF Eu Test Plate Wallac TRF Tb Test Plate Wallac TRF Sm Test Plate Wallac TRF Dy Test Plate Europium Updated Wallac TRF Eu Laser 445 Test Plate Wallac TRF Eu Laser 446 Test Plate LANCE Time resolved Fluorescence S 1536 General Europium lt Back Next gt Cancel
48. installed in your instrument Please consult the Order guide for a list of options which can be installed by a service person Introduction to the User Interface Introduction to the User Interface When you boot the system the main Reader control window appears You can start assays by clicking the Start button This begins the Assay Start wizard You can also select from a list of ready protocols and start running them with the Run button Reader control window This is the window used to control and view a run The main parts are shown in the figure Wallac nVision Manager 1 11 Simulation mode The menu title bar has six titles Introduction to the UserInterface File menu This menu allows Exit from the software Edit menu Not used in Reader control View menu You can use this to go to one of the views e g Results etc Keyboard shortcut can also be used and these are shown in the View menu If you have a tree displayed then you can move back or forward in the tree or go up to the next highest level You can also select whether you want the Shortcuts bar and or the Navigation Tree to appear Tools menu The Assay start Wizard shows you how to optimize and start running plates Before moving the instrument click Prepare Instrument for Transport Confirm the selection by clicking OK Actions menu The following functions are available under Action Latest Results display the results that have been most
49. irg Retrieve liquid back to the bottle Denan bead Prom the syringe and butler to the baud reserver This operation allows you to dispense a selected volume of liquid into the waste container You can use this operation to empty tubing if you perform it with the inlet tube in air Click Next to continue This operation allows you to rinse the tubing of the selected pump or pumps 116 Dispenser Control Select the tip mount to be used This must be the one in the instrument as shown by the mark See Show advanced options for some extra parameters you can edit e l Dispenser Control Mortenarce senna Maver rere Binwet Ehre Tomoni 5 ea tee neri D Te B7 ee abe ara end opta Tutal vohama 20010000 of pens 100400 yj Tp tube vote 50 2000 14 hrga ation uta vihane ih EESE REI EIEE Tota iute voima Ue Click Next to continue or Back to return to the Maintenance page f oot tet l Dispenser Control Tarpas artsd actvatack Cret 22 68 C Paade Click Start or press PUMP button on the dispenser to Rinse 117 Dispenser control Click Start or press the PUMP button on the dispenser Rinsing will then take place An animation will show what is happening ft simlaion mode eR AE a J S t l Dispenser Control You can repeat this operation if required to rinse with PUMP2 Click Finish The Maintenance display will reap
50. is based on the Create new protocol option with comments about the other options where relevant Note The protocol created with the Assay Start Wizard will use default parameter settings and is for one plate only You must use the protocol editor Protocols if you want to define the protocol parameters yourself 43 Using the Assay Start Wizard 3 Protocol If you are creating a new protocol you will be asked to tell in which folder the protocol is to be saved Select the folder from the tree and click Next Assay Start Wizard Create new protocol Please select the folder and name for the new protocol Click Next to continue Select protocol New Protocol 2 User protocols SE O User 2 User 3 User 4 User 5 Note If instead you are using an existing protocol for optimization or running select it from the tree Assay Start Wizard ion Select protocol Please select the protocol you want to run The wizard will jump to the last window where Select protocol you can start the assay Click Next to continue m User protocols w Assay examples O Wallac protocols Note If you select a protocol for a run but the protocol is not optimized an error message will appear and you must optimize the protocol User protocols 5 User 1 New Protocol 1 Kinetic example Copy of Standard curve fitting A Invalid protocol Selected protocol is not valid val
51. lick Next 104 Advanced optimization Measurement height optimization If you leave the Scan plate for strongest sample check box checked then the position of the sample to be used for the measurement height optimization is determined by the system lt Assay Start Wizard Plate preparation Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Scan plate for strongest sample Plate preparation 1 2 3 4 61 6 7 8 9 40 41 12 Numberof flashes eceeeeeeeeooo 0000060060000 0 If you uncheck this box you can either accept the default position which is shown in the plate map Scan plate for strongest sample Plate preparation 1 2 a 4a s e 7 s a 10 14 42 Number of flashes A 10 1 5000 8 c D E F 6 H Height sample a 105 Advanced optimization or you can show the position of the sample you want to use by clicking on the plate map using the left mouse button and selecting the word Height that appears in the tool tip Assay Start Wizard Select action Plate preparation Soler praboccl Please prepare the plate according pea Soar instrument Click Next to start the Optimization info Plate preparation Cl Scan plate for strongest sample i ie ee sa res elol ol When you click Next the optimi
52. loaded the plate either press the LOAD button again or press START to move the plate carrier and plate into the instrument The LOAD button will light up during the time the loading or unloading is occurring otherwise it will be off Order of Dual label measurement in single detector instruments In the case of single detector instruments a different procedure is followed for dual label measurements than for instruments with two detectors This is to decrease the 70 Running an assay without the Assay Start Wizard number of times the emission slide moves and to speed up the measurement of dual labels The order of measurements is e Well 1 emission slide position 2 followed by emission slide position 1 e Well 2 emission slide position followed by emission slide position 2 e Well 3 emission slide position 2 followed by emission slide position 1 e etc Note This is valid for Dual excitation labels which are always measured with one detector e g two excitation filters and one emission filter as in the case of the Fura 2 label shown with the Dispenser option in simulation mode 71 Running an assay without the Assay Start Wizard 72 Chapter 3 Result viewing 74 Result viewing Result viewing To see results click on Results in the Navigation Tree A list of assays and dates will appear Wallac Envision Manager Simulation mode File Edit View Tools Actions Help 09 0 8 e D Back Forward Up Start
53. meter determines the volume per second of liquid aspirated The default is 200 ul s The more viscous the liquid the higher the speed you should use Tip tube volume 50 2000 ul The volume of the tube between the syringe and the tip mount is known for a particular tip type and this volume appears as the default parameter value The software allows you to change this value if it is necessary Aspiration tube volume ul The value of this parameter is fixed here but can be changed under the Settings tab Total tube volume ul This is the sum of the Tip tube volume and the Aspiration tube volume If you change these parameters and want to save the changes select Remember settings Settings Select the Setting tab to see these parameters Note the following settings only take effect if you click the Set button 133 Dispenser control 11 Simulation mode DER og Out Start Latest Maintenance Settings F Stirring Speed 150 rpm Heating Temperature 25 C Set Advanced settings Pump 1 Pump 2 Connected to tip 1 g Connected to tip 2 m Aspiration speedis 70 of dispensing speed Aspiration speedis 70 of dispensing speed Aspiration tube volume 270 pl Aspiration tube volume 270 ul Stirring Select this check box and then select the speed to be used for stirring the liquid in the liquid reservoir The range is 100 to 500 revolutions per minute He
54. n to the UserInterface Note If you are using Enhanced Security there are restrictions on deletion and restoring See the Enhanced Security Administrator manual for more details Tabs in the Reader control window In the Reader control window there are four tabs Counts Graph Temperature or Events Reader control window Counts Wallac EnVision Manager 1 11 Simulation mode Fie Edt View Took Actions Help oer 3e Ooo LOR sat Latest Es Run oxi Resad W Idle Sainct protocol vad peotocots marked weh and cick Ruri button to start the assay Protocok ABS 405 Labet Absorbance 405 Source Absorbance value Y Assay 005 Plate 001 repeat 01 Courts Graph Temperature Events 1 2 3 s 6 u a ee ee ven 808268626266 0 5 Note If results have been obtained after clicking Run the intensity of the signal from each well is shown by how the well is colored If it is gray the sample has saturated the detector At the top of the window is information about the measurement The status of the instrument is shown and information is given on what you can do E g if it is Idle you can start the run by clicking Run The side bar on the right of the window gives you the following options 18 Introduction to the User Interface View Three option buttons allow you to select how you want results displayed The options are by Color Value numerically or Color and value If you select Color then the s
55. ned the links between the barcodes and protocols as described in Barcode settings 67 Running an assay without the Assay Start Wizard 1 Load the magazines into the stacker One of the magazines should have plates loaded in it and the other should be empty In Reader settings Stacker you can select which stacker is for loading plates and which for unloading Make sure the plates are orientated correctly with the Al position in the left hand corner furthest from you At least the first plate should have a barcode on it to select the protocol to be used to count it Note Check that the handles of both magazines are down during a run so that the plates can move upwards without hindrance 2 Press the START button on the instrument The plate carrier with the plate will go in to the instrument and measurement will begin During a run you can click either the Pause or Stop buttons if you need to pause or stop the measurement Pressing the STOP button on the instrument has the same effect You can follow the measurement by means of the plate map with colors indicating the intensity of the signal from each well At the top of the window there is information about the status of the measurement All the plates in the stacker will be measured if you have selected the Unlimited for the Number of plates parameter in Protocols Otherwise the run will stop when the selected number of plates has been counted 68 Running an assay
56. nformation about the status of the measurement Depending on the measurement you can select the Source of the signal e g which channel is displayed from the drop down list fluorescence polarization also has the mP value calculated 6 When the plate has been measured the plate carrier will come out and you can remove the plate If more than one plate has been defined in the protocol then load the next one 7 Press the START button on the instrument or click Run on the user interface Continue this procedure until the run stops It will stop automatically when the number of plates defined in the protocol has been reached If Number of Plates is Unlimited you have to press STOP or click Stop to end the protocol Running an assay with protocol barcodes no stacker Note Make sure you set the barcode mode in Reader settings under the Barcode tab You must select there the barcode to be read and then either select Define the protocol using or Split barcode for the protocol You must also have defined the links between the barcodes and protocols as described in Barcode settings Note In Barcode mode the Run button is not enabled 64 1 Running an assay without the Assay Start Wizard If the plate carrier is in the instrument press the LOAD button or click the Unload button on the toolbar The plate carrier will come out Note The software button shows Unload if the plate carrier is in the instrument and Load if i
57. nformation you need to run EnVision in the best way for you Do you have a protocol ready No Yes Use the Assay Start The next question is then Optimization Wizard under concerned with Tools or the Start button to optimization create and optimize one Or go to the protocol editor Protocols and create a protocol then come back to these questions Note The protocol created by the Assay Start Wizard has default parameters for a single plate and single label It allows you to do an optimization and then run a single plate assay You can edit the protocol later To start with a protocol using parameter values different from the default ones you must create it first before using the Assay Start Wizard 33 Options for running EnVision Do you have the plate and label combination optimized No Yes Use the Assay Start No protocol barcodes Wizard to do the optimization e Use the Assay Start Note Optimization is the procedure for fine tuning the parameters for your plate and label combination to get the very best results For some applications you can get satisfactory results by using the default settings without optimizing See the table Note For multiple label assays you must run the Assay Start Wizard for each label Wizard e Or select the protocol and click the Run button on the user interface Protocol barcodes and the barcode mode selected e Use the Assay St
58. nning with 70 74 Buttons on the instrument 75 C Calc 83 139 Index Calculation Adding additional 103 Calculations 90 Deleting 106 Color 20 31 Color coding 20 Color scale 22 Generic range 22 Logarithmic 22 Colors 14 Connected to tip 146 Control unit 6 Conventions used 8 9 Buttons 8 Navigation Tree items 8 Counts 88 Counts view 91 Crosstalk correction optimization 58 59 120 121 AlphaScreen 41 Enhanced luminescence 41 HTS AlphaScreen 41 Ultra Sensitive lumines 41 Curve fitting 92 Calculate unknowns 100 Curve parameters 93 Editing display parameters 97 Fitting options 95 Index Replicate points 99 Show gridlines 100 Show info frames 93 Curve fitting types 95 Curve parameters 93 Curve scale 26 Relative to greatest 26 Show points 26 D Delete 83 Detector gain optimization 39 117 Different combinations 76 77 Dispenser 4 Dispenser control 125 Animation 129 Maintenance 148 Settings 144 145 Show advanced options 127 142 Dispenser guidelines 146 Dual label measurement 77 E Edit protocol 12 Empty syringe 137 Empty tubing 148 Empty tubing with rinse 127 EnVision With stacker 4 Without stacker 5 Exit 6 Export 107 F File 11 Fill 130 147 Fill syringe 140 Filter 83 Filters 30 Fitting options 95 Flatfield correction 40 51 Fluorescence intensity 3 Fluorescence polarization 3 G Gain Detec
59. o Please select optimization s you want to run For advanced options check the Advanced mode checkbox Click Next to continue Optimization info Optimizations to run V Plate Dimension Not optimized Measurement Height Not optimized v v Detector Gain Not optimized vz Not optimized Flatfield Correction Not optimized Advanced mode Alternatively if you select Flatfield correction the other four will be disabled Assay Start Wizard Optimization info Please select optimization s you want to run For advanced options check the Advanced mode checkbox Click Next to continue Optimizations to run Optimized 26 11 2007 15 43 18 Optimized 26 11 2007 15 43 18 Optimized 26 11 2007 15 43 18 Optimized 26 11 2007 15 43 18 Flatfield Correction Optimized 26 11 2007 15 32 30 Advanced mode Note If the optimization involves the TRF laser technology there will be a TRF optimization instead of Detector Gain 47 Using the Assay Start Wizard Assay Start Wizard Optimization info Please select optimization s you want to run For advanced options check the Advanced mode checkbox Click Next to continue Optimizations to run v Plate Dimension Not optimized V Measurement Height Not optimized Vv TRE Not optimized v z Not optimized Not optimized Advanced mode Choose the optimiz
60. of wavelengths for absorption measurements The dual monochromator option allows also wavelength selection for both excitation and emission light in fluorescence intensity measurements EnVision is a very compact small footprint bench top unit with features such as shaking reading from above or below and scanning Also available are a dispenser with up to two pumps and temperature control of the instrument chamber Introduction The software is a 32 bit application running under Windows XP or Windows Vista Output can be to a file on the PC and or to a laser printer or on the network Figure 1 Wallac EnVision with a stacker Figure 2 Wallac EnVision without a stacker Introduction Software installation Note Installation must be done only by PerkinElmer service personnel Note The following instructions assume that EnVision the CAN card or optional control unit PC and printer have been connected up and switched on see the installation instructions in the Instrument manual 1 Load the software CD 2 The setup loader will start automatically This will guide you through the installation 3 When you have completed the installation you should remove the CD from the PC Start up 1 Switch on EnVision If you have a control unit switch it on This is a separate CPU used to run the instrument software In an alternative configuration the job is handled by the workstation PC without a separate control uni
61. open it Shortcut Bar Reader Control k2 Inventory It may be that the Navigation Tree contains a lot more items than you normally want to use E g if you just work with Reader control and Results you may find it more convenient to use the shortcut bar and set it to only show these two items To edit the contents of the Shortcut Bar first move the functions from the Navigation Tree to the Shortcut Bar by dragging them or selecting a function in the Navigation Tree clicking the right mouse button and selecting New Shortcut When you have the items you want in the Shortcut Bar you can use the View menu or 15 Introduction to the UserInterface right mouse button menu to close the Navigation Tree You can then just use the icons in the Shortcut Bar until such time as you need some additional function from the Navigation Tree To delete a shortcut from the Shortcut Bar right click on a shortcut and select Delete From Shortcut Bar With the right mouse button menu you can also Reset Shortcut Bar so that it goes to the default configuration Reader Control and Results or Hide Shortcut Bar You can also create other shortcut Groups e g you could have one group with shortcuts for running and viewing results and a second group connected with protocol editing You can freely Delete Group or Rename Group except if there is only one group then it cannot be deleted You can select whether you have Large icons
62. or right click the mouse to close this view Track current well checkbox select this if you want the current well to be always visible during a live display This is useful when the plate is magnified and only some of the wells appear in the live display Color scale Logarithmic if this checkbox is not selected then the scale for the signal intensity is linear The range for the scale is selected from the drop down list box If no label has been selected then this will be a Generic range If the label is selected then there is a selection of scales to choose from These go from Extra low 1 1000 to UltraHigh 1 to 100 000 000 The default range for most labels is UltraHigh and linear scale For fluorescence polarization it is Extra low and linear scale For absorbance the scale is 0 6 and logarithmic The scale that is selected is then used the next time the label is measured There is also acolor scale with sliders You can move the sliders using the mouse 20 Introduction to the User Interface The numbers by the sliders depend on the scale range chosen and the position of the slider within that range Any results that are smaller than the lower slider will appear in e g violet Any results higher than the upper slider will appear in e g red You can define the colors in Reader settings Results between the values of the two sliders will appear in a color that corresponds to the value It is also
63. otifications Full protocol information Place at end of output Plate information to include X Plate information Background information lace at beginning of plate Other options Show picture Each plate to a separate file Don t add plate number to the file name Use system list separator Clicking this button on the toolbar allows you to send results to a file and or printer The options for the format of the output are the same as described in Protocol Output 98 Chapter 4 Advanced Optimization 100 Advanced optimization Advanced Optimization These windows normally appear only if you selected Advanced in the Optimization Info window of the Assay Start Wizard Only the additional advanced windows are described here otherwise operation is as described in Assay Start Wizard In normal optimization you would not change these parameters but you may want to as part of Advanced optimization Plate dimension optimization lt Assay Start Wizard Plate preparation Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of the plate Plate preparation 6 7 e 40 14 42 Number of flashes e 10 1 5000 Size of the scanned area EES 1 9 mm 0 85 Z value target 2 high sample oO 2 low sample Height sample Gain sample ao bn
64. ots marked with and cick Puri button to start the assay Protocol Qordord ave fitting Label Aixrexen Source Charnel Assay 004 Plate 001 repast OL Counts Geach Tomeerature Everts Deseretion Completed exporting meny 4 Sharcland curve fine Comeleted exporting plate 1 repeat 1 of assay 4 Standard curve fitting to fle C Decuments and Settings Al Users Aephcation Oe Completed saving assay s Standard curve Fitting Completed ening of plate 1 repeat 1 of anya Standard carve fitting Comeleted processing assay 4 Standard curve Fitting Completed processing of plate 1 repeat 1 of assay 4 Standard curve fitting Started assay 4 Roreiond curve ting 3 97 29 Completed exporting essay ABS 405 II Completed exporting plate 1 repeat 1 of assay 3 ABS 405 to fle C Oucuments and Setunge Al Users ipgicabon DatalPerkin l 3 37 38 Rortedd exporting plate t repeat 1 of assay 3 ABS 405 to fle C Documents and Settings User shAppiication Daba PerkinEkne 3 97 97 Completed saving essay ABS 405 3 37 36 Completed saving of plate 1 repeat 1 of assay 3 ABS 405 IIM Comeleted processing assay 3 ABS P 105 Completed processing of plate 1 repeat 1 oF assay 3 ABS 405 3 37 23 Started assay 3 ABS 405 This tab shows a log of the operations performed by the instrument during a run The log is in chronological order with the most recent operation first The log is cleared when you shut d
65. own the Manager software 27 Introduction to the UserInterface Inventory Wallac EnVision Manager 1 11 Simulation mode Temperature control actwated Curent 23 13 C Paay YA Top mirror modules 412 481 gt T US Reade Settings 2 402 By Ause Trad 2 Recyde lin 2 x x2 Bottom mirror module g Aperture 304 Plate MTS Alphasoreen agentuze 5 This window shows you visually which components are installed in your EnVision These components are Filter slides individual filters are shown in their slides Mirror modules top and bottom Tip mounts Apertures From this information you can determine if you have the components installed that are necessary for the operations you wish to perform If a component is missing any protocol that needs it will not be valid i e there will not be an and a green bar beside the protocol name in the protocol list 28 Introduction to the User Interface Install any missing components and ensure you have a valid protocol before attempting to run the assay Refer to the reference manual for information about setting up protocols filters mirror modules etc Refer to the routine maintenance section of the Instrument manual for how to physically install components 29 Introduction to the UserInterface 30 Chapter 2 Operating EnVision 32 Options for running EnVision Options for running Envision The following questions guide you to the i
66. pear 118 Dispenser Control Note If any PUMP button is pressed while the dispenser is operating that operation will be terminated immediately and dispenser initialization must be done again Fill tubing This operation aspirates enough liquid to fill the tubing A small extra amount about 150 uL will be aspirated to ensure there are no air bubbles in the tubing The excess will be dispensed into the waste container Click Next when you have selected this option This operation allows you to fill the tubing of the selected pump or pumps Select the tip mount to be used This must be the one in the instrument as shown by the mark 119 Dispenser control See Show advanced options for some extra parameters you can edit l Dispenser Control Torget contol sctwated Cument 22 66 C Modd Ansis e Pumo t Bary Tonanti 5 fes Te Pred tus To E Drs advanced optione Aol neo Volume 5010000 pt 620 Volume 5010000 ph Speed 100 500 pjs 10 Speed 100 500 utis 100 Te tube wolume SO 2000 yt 200 Tee tube vaime SO 2000 ys 200 Aspe anon tube vme 27 Aspa adon tube youre uf 270 Total tube woke m Tota tube voimme of 70 Cem Diamante saenge Click Next 2a o aoa dm J Dispenser Control Click Start or press PUMP button on the dispenser to Fill tubings Cae Cms Click Start or press the appropriate PUMP button on the dispenser Tubing will be filled 120
67. possible to manually change the range by typing a value in the box and then pressing Enter Note The type and number of colors used can be changed in Reader settings Reader control window Graph IRA H AW y y YY Press Ft for help Select this view if you are doing Plate or Assay repeats Kinetic measurement or Dispense Measurement 21 Introduction to the UserInterface The side bar on the right of the window gives you the following options View Plate select this to see a picture of the plate For each well you will see a curve showing how the signal changes with time There is a drop down list where you can select the size of the plate display You will need this for numerical values and higher density plates where the values are hard to read Magnification can be up to 300 Select Size to fit if you want the software to determine the optimum size to see the whole plate Note Press Ctrl F to see a full screen view of the plate Press Esc or right click the mouse to close this view Track current well checkbox select this if you want the current well to be visible during a live display This is useful when the plate is magnified and only some of the wells appear on screen Overlaid curves before you select this click the cursor on the wells you are interested in Each time you have selected a well click the Add curve button you can also double click the well The well coordinates will be
68. r 1 11 Simulation mode tink Log Ou Rot Latest Nepagation Tree x bn Dispenser Control Temoerature contro activated Cumont 22 65 C ROY e rreccis z B renas Hortenance setings KE bwvertory amp Cepenser Control Retrieve liquid back to the bottle Tengerre control a Messeren Tedrdoger Angis Ejo Drees Yinme2z gt Sages BS Barcode Setungs Te move S Ran Tene Prete Dus Tio UG Reader settings aara a D Recre an E Show advanced options anor poset Volume S0 10000 pf Voke 5910000 yi Speed 100 500 isi 400 Speed 100 500 pai s 100 Tonte wise somone 200 Tonte whee soz 200 Rope dan tube voume yh 270 Age ion tube vre yh 270 Tota tube volume pt m Total tube volume yi m Cena ners O Ransntar saetnge Select the pump or pumps from which you want to retrieve liquid Select the tip mount to be used This must be the one in the instrument as shown by the mark See Show advanced options for some extra parameters you can edit Click Next 122 Dispenser Control n Abanager 1 11 Simulation mode Click Start or press PUMP button on the dispenser to Retrieve liquid back to the bottle Cen Cine Click Start or press the PUMP button on the dispenser Liquid will be retrieved to the liquid reservoir An animation will show what is happening Wallac EnVision Manager 1 11 Simulation mode Temperature control actwated Current 22 Martenance s
69. r desktop Copy and paste this into an email with information about the problem and send it to your service engineer Introduction to the UserInterface Select Save Screenshot To File if you want a screenshot of your user interface at any phase in its operation The file format is bmp You must give the path to where the file is to be saved About gives information about the software version Reader control toolbar Below the menu title bar is the toolbar Some of the buttons always remain the same others change according to the view selected Wallac EnVision Manager 1 11 Simulation mode Buttons that do not change Three buttons Back Forward and Up allow you to navigate in the tree structure of the Navigation Tree The same commands are found in the View menu You can also view latest results by clicking the Latest button at the top of the window The Logout button appears if you have the Enhanced Security option The Start button allows you to optimize and run plates The Assay Start Wizard guides you through these steps 12 Introduction to the User Interface Buttons that change The buttons here depend on the view selected In the Reader control view the buttons enabled are Edit Run and Load Pause Stop and Re stack are disabled See the Action menu for a description of these functions The drop down menu allows you to select the label you are going to use Labels with a green bar and an asterisk in
70. s 1 Load the stacker In Reader settings Stacker you can select which stacker is for loading plates and which for unloading Make sure the plates are orientated correctly with the Al position in the left hand corner furthest from you Note Check that the handles of both magazines are down during a run so that the plates can move upwards without hindrance 66 Running an assay without the Assay Start Wizard 2 Select the protocol you want from the drop down list of protocols 3 Use the mouse to click the Run button on the user interface During a run you can click either the Pause or Stop buttons if you need to pause or stop the measurement Pressing the STOP button on the instrument has the same effect Note If you want to run assay repeats with an undefined number of plates in the magazine i e select Unlimited then select End the assay when stacker is out of plates in Reader settings Stacker 4 You can follow the measurement by means of the plate map with colors indicating the intensity of the signal from each well At the top of the window there is information about the status of the measurement All the plates in the stacker will be measured if you have selected the Unlimited number of plates mode in Protocols Otherwise the run will stop when the selected number of plates has been counted Running an assay with a stacker and protocol barcodes Note Make sure you set the barcode mode and have defi
71. samples are in the correct layout in the plate load your plate and click Next Note In general unless you have selected Advanced you should not edit the Number of Flashes or Size of the Scanned Area even though you can do this See Advanced Optimization for information about these parameters Note If your protocol uses fluorescence polarization the Advanced check box will be selected by default If 51 Using the Assay Start Wizard Detector Gain optimization is selected you can select to use a Blank sample and give the Target mP value This is so that you can check that the S and P signal levels are in a suitable range 200 000 400 000 per flash and that they match the target mP value Assay Start Wizard Optimization info Please select optimization s you want to run For advanced options check the Advanced mode checkbox Click Next to continue Optimization info Optimizations to run Plate Dimension Optimized 27 11 2007 11 52 46 Measurement Height Optimized 27 11 2007 11 52 46 Detector Gain Optimized 27 11 2007 11 52 46 Zz Optimized 27 11 2007 11 52 46 Flatfield Correction Optimized 27 11 2007 12 01 30 Advanced mode Use Blank Correction Targeted mP value 27 0 500 mP The blank sample should be put next to the Gain sample on the cassette 52 Using the Assay Start Wizard Assay Start Wizard Plate preparation Pleas
72. se signal in the case of absorbance the signal will be lowest here Color coded contours show the signal intensity with red the most intense The order of the wells is Well upper left corner Well 2 upper right corner Well 3 lower left corner Well 4 lower right corner 103 Advanced optimization Assay Start Wizard Plate results Please check each well that the crosshair is in the center of the well and modify if necessary Click Next to continue Well well2 well well4 Plate results eae a RFU X 0 ay i 2 J 234983 Y 0 e LJ 252843 S af E 210702 E 168562 o E 126421 E 84281 E 42140 E 0 42140 O9 a 1 1 0 9 mm o 4 5 2 A 1G Al 2 E 1step 0 9 mm X Position 14 38 mm Y Position 11 24 mm 96 General amp Fluorescein Note In fluorescence intensity and fluorescence polarization measurements the signal should not exceed 500 000 RFUs for a single flash otherwise saturation of the detector may occur and you will not be able to be sure of the position of greatest intensity In this case and if the red area is not symmetrical you should point to the center of the corner well as indicated by the general shape of the contours The crosshairs should be in the centre of the well If they are not you can move them with the mouse or by clicking the Step arrows You can also select the size of the step When you are satisfied with the position for each well c
73. sition in the left hand corner furthest from you 3 Select the protocol you want to use from the drop down list of protocols Wallac EnVision Manager 1 11 Simulation mode Ee ER Yeow Jods atos Hio O Assay NfA Piste N A repest Nja 62 Running an assay without the Assay Start Wizard 4 Use the mouse to click the Run button on the user interface The plate carrier with the plate will go into the instrument and measurement will begin Note If START is pressed on the instrument the last run protocol is repeated not the one selected from the drop down list During a run you can click either the Pause or Stop buttons on the toolbar if you need to pause or stop the measurement Pressing the STOP button on the instrument has the same effect Y Wallac nvision Manager 1 11 Simulation mode HD Measur ing Processing assay meanaing wets Chk Rep button to abort the aray lop Seve Label Exrepnan Source Chine S Assay 600 Plates C01 repast OL Conte wah tenerae Everts PRS PS es ar Wd Ek Cd ee Pod Pad et Track curent wet O98 56056802 8060 Crogen 0062866620600 D 10000000 10 000000 6 000 090 6000 000 c 400000 Proce F1 for heip Sion mode aa 5 You can follow the measurement by means of the plate map with colors indicating the intensity of the signal 63 Running an assay without the Assay Start Wizard from each well At the top of the window there is i
74. stgease 12 Configuring your user interface s cs2 03 cccssececebeadscdbecastcedsnieecodietenentaeds 13 Navigati n Tree eie ien erie ates aaiae le cans dye eee cane deaets 14 Shortcut Batine osn E EAE A EA 15 JELET Cle TRIM EE A E EE E aces E eae 17 Tabs in the Reader control window ssssssesssesessseesseessseessressessseeesseeessees 18 Reader control window Counts cccccessceceeneeeeeeececeeeeecseeeeeseeeenaeeees 18 Reader control window Graph esesssesessssessesseessessssersserssseeesseesseesse 21 Reader control window Temperature sseseeeeeeereeseesreereesriseresreseee 25 Reader control Window Events cscccesseeceeeecesececeeeeeceeeeecseeeeeneeees 27 InyentOry a e a lee a a a aaa a a Sat 28 Options for running Envision sssesssssssecssecssocesocesoocssocessecesocesocesoosesoeessee 33 Do you have a protocol ready ous saxactesdasscate Soazaceds seacduatsnasthdl eaceessemacnaiaeees 33 Do you have the plate and label combination optimized eee 34 Types of optimization siciseisjcacdusvaveisiseyeaseassnszeea iesenunnsvarsceasdevecennsccaatelabeveces 35 Plateonentat om i ea casa sat ve Se Geant a a a Es 39 Using the Assay Start Wizard seeescoescoessocsssccssocesoossoosessesssccssocesocesooseso 41 ASSay Start Wizard siiu nen a asad E E RE 41 Use of an optimization plate cee eescceenececseececseececeeeeeeseeeeseeeeeaeeees 41 Steps in running the Assay Start Wizard eeeeeeeee
75. t 3 Wait until the red light on the EnVision panel is steady not blinking This takes a couple of minutes 4 Switch on the user PC 5 Start Wallac EnVision Manager by clicking the EnVision icon Caution For users with the Stacker option keep your hands away from the stacker area when the software is started or restarted The rods in the stacker will come up during the initialization process Introduction Shut down 1 To shut the loading door press the LOAD button 2 Close Wallac EnVision Manager by clicking File and 5 Exit or closing the window in the usual way You will be given a choice to shut down either the workstation software only or the whole EnVision software Choose the option you want and click OK Shut down the user PC If you have a control unit press the ON OFF button to shut it down Wait while the light is flashing on the control unit two or three minutes Only if this button has not ceased flashing after several minutes should you press the same button again Shut down EnVision Note If you are not going to use EnVision for a while you can leave the power to the various units switched on but you should shut the loading door Introduction Using this manual This manual describes the main steps to be followed when running samples on EnVision It assumes that protocols have been set up and that all the needed components filters etc have been installed Please refer to the Ref
76. t is out 2 Load your first plate Make sure the plate is orientated correctly with the A1 position in the left hand corner furthest from you The plate should have a barcode on it to select the protocol to be used to count it Press the START button on the instrument The plate carrier with the plate will go in to the instrument and measurement will begin During a run you can click either the Pause or Stop buttons if you need to pause or stop the measurement Pressing the STOP button on the instrument has the same effect You can follow the measurement by means of the plate map with colors indicating the intensity of the signal from each well At the top of the window there is information about the status of the measurement When the plate has been measured the plate carrier will come out and you can remove the plate If more than one plate has been defined in the protocol then load the next one Press the START button on the instrument 65 Running an assay without the Assay Start Wizard Continue this procedure until the run stops It will stop automatically when the number of plates defined in the protocol has been reached Note If plates after the first one have no barcodes they will be counted with the protocol defined by the first plate barcode If they have their own barcode then the protocol defined by that barcode will be used to count the plate Running an assay with a stacker no protocol barcode
77. te 17 Delete groups 17 143 Index Groups 17 Large icons 17 New shortcut 16 Rename groups 17 Reset shortcuts bar 17 Right mouse button 17 Small icons 17 Show advanced options 142 Show fitting info 91 Show points 26 Size of the scanned area 112 Size of the Scanned Area 56 Size to fit 24 Software 4 Software installation 5 Speed 144 Speed of stirring 145 Stacker 6 74 Magazines 72 74 Running with 72 74 Start 128 131 134 137 139 141 Start button 73 START button 36 72 73 74 76 Stirring 145 Stop button 69 71 73 75 STOP button 69 71 73 75 77 Strip plates 42 Switch on 6 T Temperature 27 Temperature of reservoir 145 Index Test dispense 135 Time resolved fluorometry 3 Homogenous 3 Tip Connection to pump 146 Tip mount 127 130 133 136 Tip mounts 30 Tip tube volume 144 Toolbar 13 Total tube volume 144 146 Aspiration tube volume 146 Total volume 143 Track current well 21 24 Transport prepare for 11 TRF window optimization 39 U Unlimited 74 75 Unload button 67 71 Vv Value 20 View 20 24 Both 20 Color 20 Overlaid curves 24 Plate 24 Size to fit 21 24 Track current well 21 24 Value 20 W Wavelength optimization 38 Wizards Assay Start 111 Z Z optimization 40 144
78. the plate Scan plate for strangest sample Plate preparation a2 a gt 4 5 68 7 8 9 10 11 12 Number of flashes 10 1 5000 Height sample Gain sample a AE lt gt 96 General e Fluorescein lt Back Next gt Cancel or you can show the position s of the sample s you want to use by clicking on the plate map using the left mouse button Select the sample type Height or Gain Y Assay Start Wizard Plate preparation Please prepare the plate according to the plate instrument Click Next to start the measuremer Optimization info Plate preparation Cl Scan plate for strongest sample 1 2 lle eee eae ETA e eo gt 108 Advanced optimization When you click Next the optimization will begin An additional window will appear to allow you to check and if you want to adjust the gain Assay Start Wizard Gain results Please check the gain value s and G factor and correct them if necessary Click Next to continue Channel 1 RFU Excitation Light Gain results lt gt 96 General e Fluorescein Click Next to go to the final window of the wizard 109 Advanced optimization Crosstalk correction optimization Enh lum and Ultra Sensitive lum After you have run the optimization as described in Assay Start Wizard the results will appear X Assay Start Wizard Crosstalk results Please check the calculated crosst
79. tions Number of assay repeats B s Plate 1 Start assay repeat each N A AO Group 1 Number of plate repeats 1 Fluorescein 1 Start plate repeat each N A Calculations Is label meas height used Yes 1 Curve fitting star 2 Avg of type 3 Avg of type Height of measurement Mode of measurement Rotated plate Protocol notes Defined in labe This can be dew Plate type Name of the plate type Number of rows Number of columns 12 Number of the wells in the plate Wallac Test Pla Coordinates of corners Wallac Test Plate x 113 14 m 74 06 um 0l 02 03 04 05 06 07 08 09 10 ll 12 A UNK1 UNK1 UNK2 UNK2 UNK3 UNK3 UNK4 UNK4 UNKS UNKS UNK6 UNK6 IB UNK UNK UNK8 UNKS8 UNK9 UNK9 UNK10 UNKLO CTL1 CTL2 UNK11 UNK11 C UNK12 UNK12 UNK13 UNK13 UNK14 UNK14 UNK15 UNK15 UNK16 UNK16 UNK17 UNK17 D UNK1S UNK18 UNK19 UNK19 UNK20 UNK20 UNK21 UNK21 UNK22 UNK22 UNK23 UNK23 79 Result viewing Notifications Assay 7 Results loj x File Edit View Tools Actions Help oo lali Back Forward Up Notifications Assay ID 7 Simulated Ti EF Notifications S Plate 1 5 Group 1 Fluorescein 1 HES Calculations fe 1 Curve fitting star 2 Avg of type 3 Avg of type If any error messages or warnings have been produced they will appear if you click Notifications A yellow exclamation mark shows a warning e g if the signal
80. tlier rejection you can set criteria for rejecting outliers If a point satisfies one of these criteria then it will be rejected and not included in the curve Adjust with reference curve the default selection is No but you can also select Constant or Linear In these cases the other parameters are enabled so that you can edit them if required Edit display parameters you can adjust how the curve is displayed by changing these parameters You can 88 Result viewing experiment with the options and check boxes to see which one gives you the display you want There are three tabs Display Options Plot Metrics and Difference Plot Show options Axis boundary limits Plot metrics X transformation transFormation ol me fae o meee mere poems fare X axis annotation Y axis annotation Real world Real world O Transformed O Transformed Difference plot compared to Main curve Which mait No difference plot p MEENA O Absolute difference O Relative difference Cancel In the first one the option buttons allow you to select which metrics are to be used and if a difference plot is displayed The next tab shows check boxes which determine the way the curve is displayed 89 Result viewing Display Options Plot Metrics and Difference Plot Axis boundary limits Show options for curves Show curve s Show calibrator averages Show replicate points Color and pattern when unselected Main cur
81. tor gain optimization 39 Generic range 22 Graph 23 Graph view 91 Grid view 19 23 Group 88 Counts 88 Kinetic 88 List 88 Results 88 Guidelines for dispenser use 146 H Heating 145 Help 12 13 140 About 13 Create Error Report 13 Save Screenshot to File 13 I Init button 125 Initialization 146 Installation of software 5 Instrument buttons 9 Inventory 30 K Kinetic 88 Kinetics 23 L Laser option 3 List 89 Load button 67 71 LOAD button 67 71 77 Loading plates 42 Logarithmic 22 Logout 14 Luminescence 3 M Magazines 74 Maintenance 125 Empty syringe 137 Fill 130 Fill syringe 140 141 Index Init button 125 Initialization 125 Retrieve liquid 133 Rinse 127 Test dispense 135 Maintenance tab 148 Measurement height optimization 38 115 Menu title bar 11 File 11 Help 12 Pages 11 Settings 11 Mirror modules 30 Monochromator 3 Absorbance 38 Fluorescence intensity 38 N Navigation tree 8 15 Back button 15 Forward button 15 Right mouse button 16 Up button 15 Notifications Results 87 Number of flashes 111 112 Number of Flashes 56 O Open 83 Optimization 111 Index Advanced 56 Assay notes 66 Crosstalk correction 120 121 Detector gain 39 54 117 Detector Gain 57 Flatfield 51 Flatfield correction 40 Measurement height 38 115 Monochromator 38 Number of flashes 56 Plate dimension 37 111 Plate maps 55
82. tronger the signal the more red the color is The weaker the signal the more blue it is If you select Value you will see the actual counts or absorbance values on a white background E breede Settings N Reader Settings Courts Grach Temcersture Evenes ree ra 1 2 3 4 5 6 8 9 10 it 12 2 Recyde Bn A 0 173 0 099 0355 0 325 0 957 0550 0407 0 709 0 177 0 649 0681 1246 B 043 062 0357 08S L141 0103 0203 O47 0 0 0 089 0 562 0 163 Scetore x E Track current wel CON 0 684 0 009 0 103 1 447 0 685 0409 0420 0 997 0 62 0 983 0 267 Color scale Megat D 0 133 0 609 0 198 78 0 176 0 038 0 038 0 566 9 Absorbance scale Aaaa 0060 0 828 0 243 0 161 404 6 6 F 0018 0 279 01 0 225 O15 1 10 0 086 025 Ja s09 1 G 04 1 469 10 0 9 K 0 006 0 404 0 0 H 0 473 0 468 H 0 499 0 314 0 254 126 0 079 0 4 0 03 0 153 00 48 9 0 129 0 036 0 Press F1 for help Senlstion mode ass If you select Color and value then they will appear on a colored background where the color shows the signal strength There is a drop down list where you can select the size of the plate display You will need this for numerical values and higher density plates where the values are hard to read Magnification can be up to 300 Select Size to fit if you 19 Introduction to the UserInterface want the software to determine the optimum size for seeing the whole plate Note Press Ctrl F to see a full screen view of the plate Press Esc
83. ts value is selected You can adjust the result manually if required by clicking on the wavelength you want Monochromator wavelength optimization FI This optimization determines the wavelength at which maximum emission occurs A single sample is used Select the lower and upper limits of the wavelength range Select also the step length between wavelength measurements and the number of flashes A plot of wavelength against counts is displayed and the wavelength for the highest counts value is selected You can adjust the result manually if required by clicking on the wavelength you want TRF window optimization TRF This optimization determines the best delay between the excitation flash and the opening of the measurement window It also determines the duration of the window This minimizes the background fluorescence in the measurement window while allowing the measurement to start as soon as possible and be as short as possible thus reducing the measurement time A low and a high sample are required The number of flashes to be used for the 36 Options for running EnVision optimization needs to be selected the default is 50 Two sets of results are displayed The first proposes an optimum value for the delay based on the signal to background ratio obtained The second proposes the duration of the window Three criteria are available for determining the optimum duration These come from different parameter values in the equation
84. ults window just like the one that appears when you select Open Results window When you open the results window you will see the results tree in the left pane Click on a branch to see the corresponding view Assay Assay 7 Results lo x File Edit View Tools Actions Help 90 268 N f Back Forward Up Export Calc Navigation Tree i B 7 Simulated 23 01 2007 14 5 Notifications zarazan 15 0 B s Plate 1 Peas HQ Group 1 Standard curve ID Fluorescein 1 2 Calculations 1 Curve fitting star 2 Avg of type 3 avg of type Protocol This displays information about the assay 78 Result viewing Protocol Assay 7 Results 5 x Eile Edit View Tools Actions Help ororekin Forward Up Export Simulated Instrument nickname EnVision Instrument serial number 0 Assay started 23 01 2007 14 59 Es Plate 1 N A EH Group 1 or Fluorescein 1 EH amp Calculations 1 Curve fitting star 2 Avg of type Standard curve fit Wallac Test Plate Labels This displays the parameters of the protocol used to run the assay Select either a short list with the Basic tab or a longer list with the Extended tab Assay 7 Results loj x Eile Edit View Tools Actions Help 9 0 3 ng fi Back Forward Up Export Assay ID 7 Simulated Protocol Standard curve 1 LD Notifica
85. unning The example shows the results of selecting by Protocol Wallac Envision Manager Simulation mode Eile Edit view Tools Actions Help 9 0 2 Forward Up e 6 2 BX Start Latest Filter Open Delete Results Fiter Applied Protocol FP FITC Dual I Amount show last 500 results oO I Time From 16 01 2007 00 00 00 To aayo1j2007 15 00 13 76 Result viewing Results toolbar buttons In the variable part of the Results toolbar there are three buttons Filter If you click this button once the filter options disappear leaving only the assay list Click it again and the filter options reappear Open Clicking this button when you have selected an assay from the list of assays causes the results of the run to be displayed in a separate window You can also double click on the result to open it Delete Select the results you want to delete You can select several adjacent results by holding down the left mouse button and dragging the cursor over the ones you want Hold down the Ctrl button and click individual results to select results that are not adjacent When the results are selected click the Delete button Confirm the deletion by clicking OK Note Results can be restored from the Recycle Bin provided it has not been emptied 77 Result viewing Latest The Latest button enables you to see the results of the most recent run It opens a separate res
86. ve Comparison Unsaved Adjusted curves sum curves Color and pattern when selected The last tab gives you options to select the axis boundary limits Display Options Plot Metrics and Difference Plot Show option Axis boundary limits Min Axis limits are given in annotation metrics See Max tab Plot metrics 90 Result viewing Activate Inactivate replicate points if you want to see the effect of omitting a replicate point you can do this by unchecking the box for the point Toggle Replicate Points ed Toggle Replicate Points x Replicate point l Is active Is active Replicate point Conc 5 Conc 5 Resp 66914 v Resp 66914 v Resp 279275 v Resp 279275 Conc 500 Conc 500 Resp 220400 Resp 330306 Resp 220400 Resp 330306 The unused point appears as an open square on the graph instead of a cross Assay 7 Standard curve fitting vx f X Apoy Cancel Cak4i Oclete T Mool General settings ae Patel F Wells selaction Group 1 UL Mesruremert Frucrescen t Label Coleudaticns Lem 1 Curve tating standards or sanai nes ites eee X aan trarafermation None Phucrmicmn Chanel 1 535 Yoanistraraformation Nore Reject outers C chek Achearced Par amators Edt Fitting Par arreter Concentration
87. ve name Standard Curve Fitting properties Outlier rejection Fitting LinRear Absolute difference 0 Method options Weighted From Median 100 Curve direction Indefinite Direction Indefinite X axis Transformation None From Curve 100 j Y axis Transformation None Discriminator 2 Adjust with reference curve Curve read only parameters Fitting status Fit OK Intercept 172100 Summing Count 0 Variance ratio 0 000 Fitting select from the following types Linear regression Spline in this case the Spline smooth parameter is enabled 4PL 5PL MichaelisMenten Spline smoothing this only appears if Spline is selected You can select the amount of smoothing applied to the Spline curve from simple interpolation 87 Result viewing through various percentages of smoothing to full auto smoothing Weighted fit click this check box if you want the fit to be weighted Curve direction select if the curve must go up or down or without a special direction Transformations for the X and Y axes Hyperbolic Logarithmic or None in the case of the X axis and many more in the case of the Y axis Scale factor for the X and Y axes for hyperbolic transformations select from Log 0 1 1 or 10 Curve read only parameters these cannot be changed they just tell about the Fitting status Intercept Summing count and Variance ratio Curve name You can edit the name in this field if you want Ou
88. without the Assay Start Wizard Note If plates after the first one have no barcodes they will be counted with the protocol defined by the first plate barcode If they have their own barcode then the protocol defined by that barcode will be used to count the plate Buttons on EnVision These buttons have the following functions START button This button will light up when you press it and remain lit as long as a measurement is continuing If the plate carrier is out when you press the START button the carrier will go into the instrument and the measurement will start with the last protocol that has been run If the system has just been switched on there is no previous run and no measurement is done 69 Running an assay without the Assay Start Wizard When a plate has been measured the plate carrier will come out If other plates have been defined in the protocol the START light will be on If all the plates required by the protocol have been measured the START light will be off STOP button The light on this button will be lit as long as no measurement is occurring If there is a run occurring and you press this button the run will stop and the plate carrier will come out and the STOP light will come on The raw results of the samples measured up to the time when the STOP button was pressed will be valid LOAD button Press the LOAD button to bring out the plate carrier so that you can load a plate When you have
89. zation will begin Assay Start Wizard Height results Please check the height and modify it if necessary Click Next to continue RFU 400000 300000 200000 100000 1 3 6 naan 2S 10 A 2 Height mm Height 8 2 RFU 396313 gt 96 General b Fluorescein 106 Advanced optimization An additional window will appear to allow you to manually adjust the optimum height of the plate Click the height you want to select The red line will move to that position and the height at that position will be displayed Click Next to continue Detector gain optimization Fl and FP If you leave Scan plate for strongest sample checked the position of the gain sample is determined by the system Note When you use the Scan plate for strongest sample option in fluorescence polarization it is recommended that you check with a low binding assay sample that the S and P signal levels are in a suitable range 200 000 400 000 per flash and that they match the target mP value which is shown as part of the optimization results If you uncheck this box you can either accept the default position that is shown in the plate map along with the Height sample if that optimization has been accepted 107 Advanced optimization lt Assay Start Wizard Plate preparation Please prepare the plate according to the plate map below Load the plate into the instrument Click Next to start the measurement of

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