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ImageQuant™ LAS 4000

1. Auto a5sec 1f30sec a Select Sensitivity Resolution Sensitivity Resolution V High Resolution tsh rg TIP Select Sensitivity Resolution in the Help menu You can display Help that describes the relation between sensitivity and resolution 5 SensitivityResolation 6 Click the Start button The orange Busy LED lights on the instrument and a progress dialog is displayed during exposure 58 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 9 Exposing dye stained samples and films Trans illumination Step Action Adjust the gradations of the exposed image then save and print the image Click the Complete button The display returns to the main screen 4 10 Exposing consecutively Increment The Increment exposure type enables a series of images to be exposed consecutively The data for each image is added to the previous images to provide accumulated exposure data This section describes how to perform this type of experiment Place a sample on the appropriate tray Place the tray in the ImageQuant LAS 4000 then close the door Note e Make sure that the tray is placed at the required position ImageQuant LAS 4000 User Manual 28 9607 42 AB 59 4 Operating the ImageQuant LAS 4000 4 10 Exposing consecutively Increment gt ImageQuant LAS 4000 Fle COR View Option Heb ImageQuant AS 400
2. 6 Pie Note Administrator priveleges are required in order to perform a Flat Frame calibration 7 Select the method to calibrate from the drop down list Choose which tray positions to calibrate then click Start to begin the calibration 8 Once the calibration is complete click the Close button to close the dialog The calibration plate can now be removed ImageQuant LAS 4000 User Manual 28 9607 42 AB 79 5 Software reference guide 5 Software reference guide 5 1 Exposure type This function is for selecting exposure methods There are four types of exposure methods Exposure Type Description Precision Exposes for the entire time set in Exposure Time Increment Exposes for each time set in Interval Time and accumulates images Repetition Exposes for each interval time set in Interval Time and displays images for each section Program Exposure is made for the desired time and exposure intervals A file is created automatically for each read image and the file of accumulated images is also saved automatically 80 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 1 Exposure type Precision Sets the exposure time and creates one image Increment Sets the exposure interval and creates multiple accumulated images Image 3 Repetition gt Sets the exposure Image 1 interval and create multiple images CE gt Image 2 J
3. Change filter and filter position Current Filters Select Filters 1 Through Through 2 Through Through 3 Through Through 4 Through Through 5 Through Through OK Filter down Cancel 1 Ensure that the camera head and PC are connected Turn on the instrument and the PC and start the ImageQuant LAS 4000 Control Software 2 Close the instrument door ImageQuant LAS 4000 User Manual 28 9607 42 AB 17 3 Exchanging accessory parts 3 1 Changing or installing a filter 3 Select Filter Customization in the Option menu in the ImageQuant LAS 4000 Control Software main window 4 Click the Filter down button The filter changer is lowered 5 Open the instrument door 6 Turn the filter changer so the desired position is easily accessible If a filter is already in place at this position unscrew it carefully and put it back in its cover 7 Screw the new filter into place 8 Select the appropriate filter from the drop down list and click the OK button Change filter and filter position Current Filters Select Filters 1 Through Through v 2 Through i v Through 3 Through 605040 515 Di 4 Through 510DF10 5 Through y RIES R670 S75DF20 l OK Filter down L41 Note The filter changer returns to the original position when you click the Start Focusing or Method Tray position buttons 3 2 Installing or exchanging the lens Two
4. Lens hood Lens cap G 6 Disconnect the lens cable connector by rotating the knurling ring Knurling ring WA Connector of the lens cable see A Lens release lever s 20 ImageQuant LAS 4000 User Manual 28 9607 42 AB 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 1 Removing the LAS High Sens lens 8 Rotate the lens to the left by 45 degrees The lens comes off downward Note Hold the lens with both hands so as to prevent the lens guide from being damaged 9 Take out the lens slowly along the lens guide toward you 10 Place the end cap on ImageQuant LAS 4000 User Manual 28 9607 42 AB 21 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 1 Removing the LAS High Sens lens Note e Be sure to put the high sensitivity lens sideways and lower it gently when placing it on the desk Placing the lens downward may affect the mechanical precision of the lens barrel e Make sure that the filter changer has come down far enough to make room for replacing the lens e Do not let the high sensitivity lens hang on the lens guide The lens may fall causing injury or damage e The high sensitivity lens weighs 4 5 kg Be careful when handling it 3 2 2 Installing the LAS High Sens lens 1 Close the instrument door lf the power is turned off and the filter changer is down proceed to step 2 4 Select Optio
5. Saving window image directly Edit Preference File Format tab Select an 8 bit Color TIFF for mat Changing the temperature Edit Preference Set the temperature setting of CCD CCD Cooling tab Changing the folder of file Edit Preference Specify the folder where a file saving Root Folder tab is saved f re o Edit Preference Enter size in percent Fine adjusting the print size Print Adjust tab Displaying saturated data in red View Paint saturated data Red Select and check Displaying image in Negative Gray View Negative Gray Select and check Displaying image in Positive Gray View Positive Gray Select and check Creating CCD Calibration file Option CCD Calibration Select and set conditions for Start Creating Flat Frame Option Flat Frame Calibration Select and set conditions for Start Checking software version in Help About ImageQuant LAS Select formation 4000 Control Software Checking the relation between Help Sensitivity Resolution Select sensitivity and resolution ImageQuant LAS 4000 User Manual 28 9607 42 AB A 3 Detection Reagents and corresponding settings Note Classification Chemiluminescence A Appendix A 3 Detection Reagents and corresponding settings The RGB module may be used instead of the separate red green and blue Epi lights Reagent name ImageQuant LAS 4000 setting Me
6. Exposure Type 3 Increment Y Interval Time 4 Manvel w 4 Y 5 Sensitivity Resolution Repeat Limit 6 LJ Set umt Step Action Click the Method Tray position button Result The Method Tray position dialog is opened Method Tray position Method Aer Chemiluminescence Through Fluorescence a v 15150 O Digitization Epi illumination White Epi illumination Through Trans illumination White Trans illumination Through 105 x 70mm L f OK Cancel Select the Method and Tray Position Click the OK button 60 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 10 Exposing consecutively Increment Step Action Click the Focusing button Result The focusing controls appear eh sant LAS 4000 Fie EGR Vew Option Help ImageQuant LAS 4000 Exposure Type Interval Time Repeat Limit Set Lette 10 times 1 100 Flat frame Auto Confirm the sample position and focus Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Increment for Exposure Type Exposure Type 3 In Precision Repetition manui _ Program 10 Select Interval Time unit and enter a numeric value Interval Time Manual ko O 4 Sensitivity Res maar encore Repeat Limit CO Set Limit Note Interval Time can be entere
7. Select an exposure time from the drop down list or enter the exposure time manually The exposure time can be set from 0 01 seconds up to 30 hours Exposure Type Exposure Time Auto Manual 1min 1sec 12sec 14sec Flat frame 1 8sec Auto 1 15sec S 1 30sec 1 60sec 1 100sec ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 5 Exposing chemiluminescent samples Step Action Select Sensitivity Resolution Sensitivity Resolution High Resolution Add Di High Super Ultra High Binning Note The check box for Add Digitization Image can be checked to also perform a white light exposure for example to image dye stained molecular weight markers In this case the white epi light should be in place Leave unchecked if this is not nec essary TIP Select Help Sensitivity Resolution This opens the following diagram that de scribes the relation between sensitivity and resolution Click the Start button Result The exposure is started The orange Busy LED lights on the instrument during exposure ImageQuant LAS 4000 Exposure Type Predsion v Exposure Time Mto Manual w v Sensitivity Resolution P Exposure Tie 3er Finishing Tine AM0 Expos ImageQuant LAS 4000 User Manual 28 9607 42 AB 41 4 Operating the ImageQuant LAS 4000 4 5 Exposing chemiluminescent samples St
8. TIP Exposure information is printed together with the image except for the Easy Print option This is convenient since it is unnecessary to note the exposure information and the conditions The following exposure information is included Exposure date camera serial number lens type cooling temperature at the time of exposure exposure type sensitivity exposure time or interval time name of Method tray position number values at both ends of the range scope values at both ends of the gradation type of gradation Control Software version number Nov Wed 25 15 25 14 2009 Serial 1234567 FUJINON VRF43LMD Cooling 25C Precision Standard 1sec Chemiluminescence Tray Position 1 Range 512 4607 Gradation 512 4607 Linear ImageQuant LAS 4000 mini 1 1 0 17 Image 4 13 Saving exposed images The exposed image can be saved using the Save button displayed in the post exposure state or by File Save This section describes how to save an image Action 1 Click on the Save button or select File Save 70 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 13 Saving exposed images Step Action Save Function When the Save Function screen is displayed select one of the following 3 saving methods to save the photographed image Click Next button Save selected images O Save displayed images Make integrated image Save passed images Up to
9. Auto Range Gel File Format gel e Gel File Format gel Linear 16 bit grayscale encoding Can include additional information compared to tif format e Original Image TIFF File Linear 16bit Gray tif This is a 16 bit TIFF format e Window Image TIFF File 8bit Color tif This is a TIFF format that has an eight bit color gradation The changed gradation can be saved directly ImageQuant LAS 4000 User Manual 28 9607 42 AB 97 5 Software reference guide 5 10 Edit menu FlatFrame Mode ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer Auto Range Auto Manual Option Description Auto Optimal correction data is automatically set Manual Correction data can be selected in the Flat Frame selection screen Launch Applicaton This option can be used to automatically launch an external application when an image is saved ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer Auto Range C Activate save and launch function 98 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 10 Edit menu To define an application to launch do the following e Select the application to be launched from the drop dow
10. dragging the mouse plied to all images 2 The gradation conversion 5 Return to initial setting curve can be toggled between Linear or Sigmoid 3 The gradation can be changed by dragging the mouse 68 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 11 Exposing repeatedly Repetition Action Adjust the gradations of the exposed image then save and print the image Click the Complete button The display returns to the initial screen Note Only the last 100 images can be kept Previous images are discarded 4 12 Printing exposed images The exposed image can be output as a full scale print or screen print the window displayed on the screen The print can be done using the Print button displayed in the post exposure state or the File Print This section describes how to print an image Step Action For a screen print first display the window you wish to output 1 Click the Print button or select Print from the File menu Select the output setting for the printer and the type full scale print paper size print or screen print Click the OK button Full scale print 3 Paper size print Screen print The Print dialog is displayed Choose settings for the printer and click the Print button The image is printed ImageQuant LAS 4000 User Manual 28 9607 42 AB 69 4 Operating the ImageQuant LAS 4000 4 12 Printing exposed images
11. A 4 Main specifications Power requirements Input voltage 100 240 V Voltage variation 10 Phase Single phase Power frequency 50 60 Hz Rated input current 3 0 1 5 A AC power cable Use the cable supplied with the instrument Specifications of cables required for the use of ImageQuant LAS 4000 Voltage Plug connector Cable Power supply cord length Voltage Plug connector Cable Power supply cord length 100 120 V 125 V AC 13 A SJT3 x 16AWG 60 C Maximum 3 m 200 240 V 250 VAC 10A CENELEC OC 3 x 1 0 mm 70 C Maximum 3 m ImageQuant LAS 4000 User Manual 28 9607 42 AB 129 A Appendix A 4 Main specifications Installation conditions 130 Placement conditions Free space required around ImageQuant LAS 4000 Secure space for maintenance work as follows Front 600mm Right 600mm Left 600 mm Rear 200mm Top 1000 mm Table strength The allowable load must be 981 N m 100kg m or greater Other conditions 1 Decide on an installation location taking into consideration the work flow and ancillary facilities to be used 2 Required construction work and electricity air conditioning work must be complet ed in advance 3 It is not desirable to have a heat source on the IDX right side face where there is an air intake fan even if the environmental requirements have been met 4 Donotinstall the equipment near a window to avoid
12. Exposure Exposure Exposure Exposure Image 13 Image 14 Image 15 Image 16 Wait Wait Wait L Wait Exposure Exposure Exposure Exposure File name File Format Gel File Format gel Y Save Folder C Documents and 5 ant LAS 4000 Image Select Save As Template ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 3 Add digitization image 83 5 Software reference guide 5 4 Program settings Description Description 1 A template file is selected Ifa new template 7 Select the image file format to be saved is to be created select New Template Note Two types of format can be Note When the selected template selected the GE Healthcare is modified Modified will be gel format gel and a linear displayed 16 bit grayscale tiff file for mat tif 2 Set exposure intervals This is the time from 8 Close the settings window the shutter closure until the start of the next exposure Note The wait time can be set within the range of 0 seconds to 30 hours 3 Set exposure time 9 Enter a file name to be saved Note This can be set within the range of 0 01 seconds to 30 hours 4 Save the template 10 Enter a comment 5 Settings will return to the initial settings 11 Select the number of frames between 1 and 16 to be preset 6 Folder in which the template is saved 12 Select detection sensitivity Delete a template
13. Select accept the terms in the license agreement and click the Next button ImageQuant LAS 4000 User Manual 28 9607 42 AB 6 Installing ImageQuant LAS 4000 Control Software 6 2 Install ImageQuant LAS 4000 Control Software under Windows xP Step Action 5 Select destination folder in the dialog ie ImageQuant LAS 4000 InstallShield Wizard Destination Folder Click Next to install to this Folder or click Change to install to a different Folder Install ImageQuant LAS 4000 to C Program Files lt Back f Next gt Cancel e Click the Next button to install the software at the default folder C Program Files e Click the Change button to install to a different folder 6 Click the Install button in the installation dialog ii Click the Finish button 8 Start the ImageQuant LAS 4000 Control Software This will ensure that necessary folders and files are created 6 3 Install ImageQuant LAS 4000 Control Software under Windows Vista Before you begin Log in using a Windows account with administrator privileges Install the USB Control Driver Windows Vista Note During software installations you may be asked to confirm your actions in a dialog with the text Windows needs your permission to continue Enter an administrator password if prompted then click Continue to proceed with the installation Step Action 1 Disconnect ImageQuant LAS 4000 Control Software from the computer 2 Open the control
14. 1 30sec 1 60sec 1 100sec Select Sensitivity Resolution Sensitivity Resolution LJ High Resolution Standard High Super Ultra High Binning Super Binning Ultra Binning TIP Select Sensitivity Resolution in the Help menu You can display Help that de scribes the relation between sensitivity and resolution 5 SensitivityResolution 6 Click the Start button The orange Busy LED lights on the instrument and a progress dialog is displayed during exposure 50 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 7 Exposing fluorescence Epi illumination Step Action Adjust the gradations of the exposed image then save and print the image TEETE ImageQuant LAS 4000 Click the Complete button The display returns to the main screen 4 8 Exposing dye stained samples and films White Epi light This section describes how to expose dye stained samples using white Epi illumination For some applications such as imaging gels stained with Coomassie Blue dye results may not be optimal using the white epi light If the White trans tray is available see Section 4 9 Exposing dye stained samples and films Trans illumination on page 55 for a more optimal method Place a sample on the Epi tray Put the tray in the ImageQuant LAS 4000 then close the door Note e Prior to exposure click on the Digitiza tion
15. 4000 4 Operating the ImageQuant LAS 4000 This chapter describes the ImageQuant LAS 4000 Control Software and how to expose gels membranes or films to obtain digital images 4 1 Layout of the ImageQuant LAS 4000 Control Software The exposure of samples in the ImageQuant LAS 4000 the setting of exposure conditions and the viewing of the digitized images are performed within the ImageQuant LAS 4000 Control Software The following illustration shows the features of the main window of the ImageQuant LAS 4000 Control Software No Description Description 1 Menu bar The setting state such as the temperature condition of CCD is displayed 2 Exposure Type The state of the Intelligent dark box IDX is Sets the exposure method displayed 3 Exposure Time Method Tray position Sets the exposure time Sets the detection method and tray position 32 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 1 Layout of the ImageQuant LAS 4000 Control Software No Description No Description 4 Sensitivity Resolution 9 Focusing Sensitivity can be set Adjusts the focus and position 5 Add Digitization Image 10 Start A digitization image is exposed simultaneous Starts the exposure ly with a chemiluminescence image Chemiluminescence method only Note The software needs to be started once after installing from an account with administrator priveleges
16. All This function toggles the quality of a focus image The image is displayed in high image quality when this option is selected ImageQuant LAS 4000 User Manual 28 9607 42 AB 103 5 Software reference guide 5 11 View menu Pan Focus Fast This function toggles the quality of a focus image With this option selected the image quality is lower than using Pan Focus All but it is displayed faster TIP During sample alignment a sample can be quickly aligned when Pan Focus Fast is selected 5 12 Option menu 5 12 1 CCD Calibration This function creates a correction file that is used for correcting characteristics of the CCD detector The created correction file varies depending on the sensitivity exposure time short or long and CCD cooling temperature 1 When CCD Calibration is selected in the Option menu the following window is shown 5 3 1 CCD Calibration Currant Temperature 4 from 1 100 sec to 2 hours over 2 hours High Resolution Fi Ready Thu Jul 2 14 50 16 2009 Standard Oo pea Thu Jul 2 14 51 31 2009 Ready High 0 Thu Jul 2 14 52 31 2009 Ready S m Thu Jul 2 14 53 31 2009 o Da Thu Jul 2 14 54 31 2009 Total Time for Calibration 0 minutes Not Ready No Description 1 Click here to check the boxes for all resolutions for up to 2 hours 4 Click here to check the boxes for all No Description resolutions for more than 2 hours 2 The time re
17. DAPI E vy Eu F2 8 Digitization Epi illumination J white Epi white O Through F2 8 Digitization Trans illumination J white Trans white C Through F2 8 bd To delete a method select the method to be deleted and click Delete Note Only user defined methods can be deleted e To edit a method select the method to be edited and click Edit Note Only user defined methods can be edited e To adda method click Add 106 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 12 Option menu 5 12 3 Method Setting Result The following dialog is displayed Method Setting i TomsMethod x oO Blue Epi Blue r7 iH F0 85 1 Enter a Name for the method 2 Select appropriate settings for Light Filter and Iris 3 Click OK Note Optical characteristics can be corrected using the Flat Frame Calibration function for the added Method See Section 5 12 2 Flat Frame Calibration on page 105 5 13 Help menu User Manual Opens the ImageQuant LAS 4000 User Manual in the default pdf browser Getting Started Opens the Getting Started with ImageQuant LAS 4000 manual in the default pdf browser End User License Agreement Opens the End User License Agreement in the default pdf browser Sensitivity Resolution The figure showing the relation between sensitivity and resolution is displayed Click to close the window Sensitivity Resolution ImageQuant L
18. FO 85 Cy3 Green 520nm Epi 575DF20 F0 85 Cy5 Red 630nm Epi R670 F0 85 Dy781 NIR 710nm Epi IR785 F0 85 DAPI UV 365nm Epi L41 F2 8 92 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 8 Method Tray position Method Filter Digitization Epi illumination White Epi illumination Through Trans illumina White Trans illumina Through tion tion Note The method that can be used varies depending on the state of the light source set in IDX Unusable options cannot be selected Note Before clicking the OK button be sure to close the IDX door Note Filter position 1 is dedicated for chemiluminescence Do not put a filter in it Tray position The tray position is changed according to the sample size The tray size that can be read varies depending on the tray position Position Width mm Length mm 1 105 70 2 144 96 3 180 120 4 210 140 250 wide view lens 250 wide view lens rT 250mm 250mm SSS S A 210mm 140mm Position 1 Maximum exposure size when a wide view lens is used TIP e The display on the main window changes to reflect the current choice of method and tray position e The NP tray is to be used at tray position 3 using the Chemiluminescence method and the NP check box should be checked The tray will not be detected otherwise ImageQuant LAS 4000 User Manual 28 9607 42 AB 93 5 Software reference
19. LAS 4000 Control Software is started In this case do not turn off the ImageQuant LAS 4000 However if the instrument will not be used for a long period it should be turned off 3 Shut down the computer 4 Switch the ImageQuant LAS 4000 off using the power switch 4 15 Exposing large sample using wide view lens A larger sample of up to 250 x 250 mm can be exposed by replacing a high sensitivity lens with a wide view lens This section describes how to expose large samples Action 1 Install the wide view lens See Section 3 2 Installing or exchanging the lens on page 18 ImageQuant LAS 4000 User Manual 28 9607 42 AB 73 4 Operating the ImageQuant LAS 4000 4 15 Exposing large sample using wide view lens Step Action Place a sample on the tray that you selected according to the light source used Put the tray in IDX and close the door 3 ImageQuant LAS 4000 File Edit View Option Help ImageQuant LAS 4000 Exposure Type Precision Y Exposure Time O Auto Manual isec Y Sensitivity Resolution Standard Y 2 Add Digitization image ON Flat frame Auto a g Method Tray position L Focusing Psn 74 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 15 Exposing large sample using wide view lens Step Action Click the Method Tray position button Method Tray position Method Filter Chemilumine
20. LAS 4000 Preference CCD Cooling Root Folder File Format Auto Range selection in Repetition Increment mode FlatFrame Mode Launch Application Print Adjust None d Increment Digitization Epi illumination Timer Auto Range Semiautomated d Automated ld An image whose gradations are adjusted automatically is displayed when exposure type is Increment or Repetition e None 102 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 10 Edit menu An image is displayed without adjusting its gradations e Semiautomated An image is displayed with its light and shade highlighted e Automated An image is displayed with its light and shade more highlighted The effect is the same as when exposure type is Precision 5 11 View menu Paint saturated data red This function toggles whether to display saturated data in red Negative Gray This sets the display color of an image such that absorbing bands are darker and the background lighter Positive Gray This sets the display color of an image such that absorbing bands are lighter and the background darker Red This sets the display color of an image such that illuminated regions are displayed in red Green This sets the display color of an image such that illuminated regions are displayed in green Blue This sets the display color of an image such that illuminated regions are displayed in blue Pan Focus
21. LAS 4000 User Manual 28 9607 42 AB 15 2 The ImageQuant LAS 4000 2 4 Parts and accessories Green Epi light 520 nm set of two Red Epi light 630nm set of two RGB module Epi lights set of two UV Epi light 365 nm set of two NIR Epi light 710 nm set of two Filters Y515 filter Y515 GFP detection filter 510DF10 GFP EtBr detection filter 605DF40 EtBr Cy3 detection filter 575DF20 Cy3 Cy5 detection filter R670BP Cy5 Alexa Fluor 750 detection filter IR785 Alexa Ultraviolet protection filter L41 UV Note e The combined accessories and light source vary depending on the system that you purchase e Make sure to retain the caps and filter cases e Store the incident Epi light sources in a container box e The UV trans tray will degrade after many UV illuminations It can be used up to approximately 1000 times if each exposure time is 1 second New trays can be purchased separately 16 ImageQuant LAS 4000 User Manual 28 9607 42 AB 3 Exchanging accessory parts 3 Exchanging accessory parts 3 1 Changing or installing a filter Optical filters are installed in the filter turret localted under the lens inside the IDX This section describes how to install or change a filter on the filter turret Install a filter in the filter turrent and register it in the ImageQuant LAS 4000 Control Software as follows amp ImageQuant LAS 4000 File Edit View Option Help ImageQuan 3 LAS 4000
22. Navigate to the folder chosen in step 6 above and delete the template file The default locations are as follows e For Windows XP C Document and Settings All users Application Data GE Healthcare lmageQuant LAS 4000 ProgramTemplate e For Windows Vista C GE Healthcare lmageQuant LAS 4000 ProgramTemplate 5 5 Focusing The Brightness function enables adjustment of the brightness of the Focusing screen and the Adjust function enables adjustment of the focus when the LAS High Sens lens is used 84 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 5 Focusing 3 ImageQuant LAS 4000 Fie Edit View Option Help ImageQuant LAS 4000 Exposure Type Exposure Time Auto Manual Sensitivity Resolution High Resolution Y Flat frame Auto No Description Description 1 Click on the image to expand or reduce the 4 Coarse adjustment of focus display 2 The view of the focusing screen becomes 5 Fine adjustment of focus darker 3 The view of the focusing screen becomes lighter TIP Select Pan Focus Fast in the View menu The focusing speed then increases This is effective for the alignment of a sample see Pan Focus Fast on page 104 Note e When the NP tray is used adjust so that the bottom ofa plate is focused Adjustment using a printed paper facilitates focusing e Check the tray positions to be exposed in advance 56 Start Exposure starts
23. R670 DAPI E o i Digitization Epi ilumination J white Epi white O Through Digitization Trans illumination J white Trans White CQ Through Delete Edit Cancel Select an existing method to change and click the Edit button or to create a new method click Add The Method Setting edit dialog opens Method Setting 3 Name TomsMethod Light JJ Blue Epi Blue Fiter R720 Iris F0 85 i Select the appropriate Light Filter and Iris edit the Name for the method and click OK Once the method is added to the list of methods click OK to return to the main window 78 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 16 Creating a new method and performing flat frame calibration Step Operation Insert the calibration plate appropriate for the light source into the ImageQuant LAS 4000 Light source Calibration plate Red Epi light Cal plate GR pink Green Epi light Cal plate GR pink Blue Epi light Cal plate FL green 5 RGB module Use the Cal plate that corresponds to the chosen wavelength red green or blue NIR Epi light Cal plate GR pink UV Epi light Cal plate FL green White Epi light Cal plate DI white Close the door of the instrument Note Ensure the the appropriate lights and filter are in place To perform a Flat Frame calibration select FlatFrame Calibration in the Option menu The FlatFrame Calibration dialog opens
24. This is to ensure that necessary folders and files are created 4 2 Preparation for exposure Analyzing PC Step Operation il Turn on the ImageQuant LAS 4000 computer and peripheral equipment Start up ImageQuant LAS 4000 Control Software ImageQuant LAS 4000 ImageQuant LAS 4000 is in self diagnosis A message is displayed until the ImageQuant LAS 4000 is ready FEENS 2 AAAA Note The power switches of the instrument and PC can be turned on in any order ImageQuant LAS 4000 User Manual 28 9607 42 AB 33 4 Operating the ImageQuant LAS 4000 4 2 Preparation for exposure Step Operation Wait until the CCD has reached the preset cooling temperature and is ready for use ImageQuant LAS 4000 Fie ER Vew Option Hep ImageQuant LAS 4000 Exposure Type Exposure Time O ato O Manual Sensitivity Resolution Add Digitization image 3 Clow Fi Method Tray position TIP e The instrument will be ready in a few minutes The power LED on the instrument is lit blue when the instrument is ready Method Tray position and Focusing can be prepared even if the CCD has not cooled completely e The Start button can be clicked even when the CCD is not completely cooled with the EtBr trans method 4 3 Calibration Ensure that the ImageQuant LAS 4000 is properly calibrated See Section 5 12 1 CCD Calibration on page 104 and Section 5 12 2 Flat Frame Calibration on
25. be applied to all images No Description No Description 1 The range can be changedby 4 The changed gradation is ap dragging the mouse plied to all images 2 The gradation conversion 5 Return to initial setting curve can be toggled between Linear or Sigmoid 3 The gradation can be changed by dragging the mouse ImageQuant LAS 4000 User Manual 28 9607 42 AB 63 4 Operating the ImageQuant LAS 4000 4 10 Exposing consecutively Increment Action Adjust the gradations of the exposed image s as described above then save and print the image s 8 Click the Complete button The display returns to the initial screen Note Only the last 100 images can be kept Previous images are discarded 4 11 Exposing repeatedly Repetition The Repetition exposure type enables a series of images to be exposed consecutively Each image is recorded separately and data is not accumulated This section describes how to perform this type of experiment Place a sample on the tray that you selected according to the type of a sample Put the tray in the ImageQuant LAS 4000 then close the door Note e Make sure that the tray is placed at the required position gt ImageQuant LAS 4000 Fle tat vew Opon Hep ImageQuant AS 4 64 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 11 Exposing repeatedly Repetition Step Action Click the Metho
26. gt Image 3 Program gt Desired exposure time image 1 and exposure interval are set and multiple gt images are created Image 2 Exposure interval Wait gt preset in Image 2 Exposure interval Wait gt preset in Image 1 Exposure interval Wait mage 4 preset in Image 3 KON a lt P G File only Exposure interval Wait preset in Image 4 Exposure time 5 2 Exposure time This function is for setting the exposure time When exposure type is precision e Auto The exposure time is automatically calculated by histogram analysis ImageQuant LAS 4000 User Manual 28 9607 42 AB 81 5 Software reference guide 5 2 Exposure time Manual Select a suitable exposure time from the drop down list Alternatively select a time unit and type in an exposure time Exposure Time O Auto Manual 10 hr Sensitivity 5 v sec High F 15hr 2hr 60min 30min 10min Smin 7 2min g Chemiluminesce 1min 1sec 1 2sec 14sec Flat frame 1 8sec Auto 1 15sec 1 30sec 1 60sec 1 100sec When exposure type is increment or repetition Choose a time unit from the drop down list and enter an interval time Interval Time Manual 10 Sensitivity Resi cae TIP The interval time can be set from 10 seconds to 2 hours For a UV transmitting light source the interval time can be set from 10 seconds to 1 minute Note An image can be exposed for up to 30 hours in Precision mode However the image
27. guide 5 8 Method Tray position Manual flat frame mode In order to be able to manually specify a Flat frame file for the method and tray position set FlatFrame Mode to Manual in Edit Preference Click Next in the Method Tray position dialog to select the appropriate Flat Frame Correction ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer Auto Range Manual TIP Only Flat Frame files that satisfy the currently selected exposure conditions are displayed in the Flat Frame Correction list To create a Flat Frame file see the description of the Flat Frame Calibration function Section 5 12 2 Flat Frame Calibration on page 105 Note Before clicking the OK button be sure to close the instrument door 5 9 File menu Page Setup Opens the settings for the printer for example the direction of paper Print Outputs an image to a printer The image can be printed as a full scale print or screen print For the output method see Section 4 12 Printing exposed images on page 69 94 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 9 File menu Save Saves a file The default setting for the file format to be saved is that selected in Preference in the Edit menu See Section 5 10 Edit menu on page 95 Quit Exits ImageQuant LAS 4000 Control Software 5 1
28. if the warranty period has not yet expired e Problems caused by incorrect usage and or by any products other than those authorized by GE Healthcare and or problems caused by other equipment e Problems and or damage due to moving transport and or falling 8 2 Repairs 1 Before asking for repairs refer to the troubleshooting section in Getting Started with ImageQuant LAS 4000 2 Ifthe problem persists fill in the service report fax sheet at the end of this manual and contact your GE Healthcare representative 3 The guaranteed repair service period of this product is 5 years from the termination of sales Thereafter repair service may not be provided if repair parts become out of stock 120 ImageQuant LAS 4000 User Manual 28 9607 42 AB A Appendix Appendix A Appendix A 1 Glossary CCD Calibration Electric charges are accumulated on the CCD even if it is not exposed to light The accumulated electric charge is called a dark current noise The process for eliminating the noise component is performed by CCD calibration The image constituted by the dark current noise is called CCD calibration data An image that is constituted by only the electric charges accumulated during exposure can be created by subtracting the CCD calibration data from the exposed image Flat Frame In an exposure system using a 2D sensor such as a CCD unevenness due to the lens or lighting system appears on the image By correcting the unevenness an obj
29. is down proceed to step 4 2 Select Option Lens Exchange LAS 4000 fem Help CCD Calibration FlatFrame Calibration FlatFrame Calibration Pro Filter Customization Lens Exchange Method Setting ImageQuant LAS 4000 User Manual 28 9607 42 AB 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 3 Removing the wide view lens A message is displayed Lens Exchange Close the door and click OK ImageQuant LAS 4000 control software will shut down Do you want to continue Click OK 5 Lens Exchange Switch off ImageQuant LAS 4000 instrument and change the lens Turn off the instrument using the power switch 4 Remove the F mount adapter and lens while pushing the release button provided on the lens Remove the wide view lens together with the F mount adapter by rotating it Remove the lens hood from the lens Place the lens cap on co N DW WM Place the end cap on the lens and the cap on the F mount adapter 3 2 4 Installing the wide view lens 1 Close the instrument door If the power is turned off and the filter changer is down proceed to step 4 2 Select Option Lens Exchange LAS 4000 fee Help i ccD Calibration n FlatFrame Calibration FlatFrame Calibration Pro Filter Customization ad Lens Exchange Method Setting ImageQuant LAS 4000 User Manual 28 9607 42 AB 25 3 Exchanging accessory parts 3 2 Installing or exchangi
30. left sides other wise an incorrect image will be obtained e Do not insert any connector other than those for the Epi light sources ImageQuant LAS 4000 User Manual 28 9607 42 AB 13 2 The ImageQuant LAS 4000 2 3 Connections USB cable Analysis PC Note Ej ImageQuant LAS 4000 USB cable Connects ImageQuant LAS 4000 and the analysis PC Read data will be sent from ImageQuant LAS 4000 to the analysis PC through the USB cable USB 2 0 is supported Connect one end with the Image Quant LAS 4000 camera head USB connector and the other to a USB port of the analysis PC Use the USB cable supplied with the equipment e Do not connect the instrument and the PC via a USB hub e Donotconnect USB equipment other than ImageQuant LAS 4000 to the PC Simi larly do not use other USB equipment connected to the PC while ImageQuant LAS 4000 is in use This may result in the loss of image data e The PC must be certified according to UL60950 1 UL listed and IEC60950 1 2 4 Parts and accessories Epi tray F0 85 43mm LAS high sensitivity lens ImageQuant LAS 4000 User Manual 28 9607 42 AB 2 The ImageQuant LAS 4000 2 4 Parts and accessories w Cal plate GR Pink Cal plate DI Cal plate FL Green White trans tray e Gel sheet White light table UV transilluminator F1 8 24mm wide view lens F mount adapter ImageQuant
31. lenses can be used with the ImageQuant LAS 4000 an F0 85 43mm LAS High Sens lens and an optional F1 8 24mm wide view lens The wide view lens can image samples up to 250 x 250 mm in size For all other applications the LAS High Sens lens is recommended This section describes how to install or exchange the lenses ImageQuant LAS 4000 User Manual 28 9607 42 AB FO 85 43mm LAS High Sens lens F1 8 24mm wide view lens 3 2 1 Removing the LAS High Sens lens 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 1 Removing the LAS High Sens lens 1 Close the instrument door If the power is turned off and the filter changer is down proceed to step 4 2 Select Option Lens Exchange LAS 4000 fee Help CCD Calibration FlatFrame Calibration FlatFrame Calibration Pro Filter Customization Lens Exchange Method Setting at A message is displayed Lens Exchange Close the door and click OK ImageQuant LAS 4000 control software will shut down Do you want to continue 3 Click OK 4 Lens Exchange Switch off ImageQuant LAS 4000 instrument and change the lens Turn off the instrument using the power switch ImageQuant LAS 4000 User Manual 28 9607 42 AB 19 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 1 Removing the LAS High Sens lens 5 Loosen and remove the lens hood then place the lens cap over the lens
32. page 105 4 4 Placing the sample This section describes how to choose an appropriate sample tray and how to place the sample for exposure 1 Select a sample tray suitable for the type of exposure to be performed Detection Sample type Chemiluminescence Membrane Epi tray Bioluminescence Titer plate NP tray 34 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 Detection Sample type Tray Fluorescence Gel UV Trans illumination UV trans tray Gel Epi illumination Epi tray Membrane Epi tray Digitization Membrane Epi tray Gel Coomassie silver stain White trans tray 2 Place a sample on the sample tray For Epi tray a Place the sample directly on the tray For UV or White Trans trays a Cut out a gel sheet slightly larger than the sample size b Place the gel sheet on the Trans tray Lf Gel sheet c Place the sample on the gel sheet y Sample Gel sheet 4 4 Placing the sample Note The gel sheet can be used repeatedly After use wash it with mild detergent rinse with water then dry well A gel sheet can be reused about 20 times ImageQuant LAS 4000 User Manual 28 9607 42 AB 35 4 Operating the ImageQuant LAS 4000 4 4 Placing the sample 3 Choose the exposure size and tray position For Epi and Trans trays e The readable area varies depending on the tray position Place the sample in position according to it
33. panel and click Classic View in the upper left corner 3 Open Add Hardware ImageQuant LAS 4000 User Manual 28 9607 42 AB 113 114 6 Installing ImageQuant LAS 4000 Control Software 6 3 Install ImageQuant LAS 4000 Control Software under Windows Vista Step Action 4 5 In the Add Hardware dialog click the Next button Select Install the hardware that I manually select from a list Advanced and click the Next button Select Show All Devices and click the Next button From the list below select the type of hardware you are installing If you do not see the hardware category you want click Show All Devices Common hardware types Show All Device 5 Display adapters G IDE ATAVATAPI controllers IEEE 1284 4 compatible printer IEEE 1284 4 devices IEEE 1394 Bus host controllers Z Imaging devices IF Infrared devices MB sors Canta Estender fy Gee Click the Have Disk button Insert the ImageQuant LAS 4000 Control Software CD and click the Browse button Select to install the driver from the ImageQuant LAS 4000 Control Software CD ocate Look in DVD CD RW Drive D ImageQuant LAS oily ofr m E Recent Items fe Desktop temodif Location i a oY Network ai Recent Places admin Public p amp i Computer bj Floppy Disk Drive A Desktop Local Disk C db Windows j System32 prti e N a STORE N GO E f S Computer K lt Network File nam
34. quality and quantitative performance are only guaranteed for a maximum of 2 hours 5 3 Add digitization image Add Digitization image v ON A chemiluminescence and a white light epi illuminated image are exposed by single click operation when this function is set to ON After exposure both chemiluminescence and white epi illuminated images can be saved This function can be used only when the method is chemiluminescence 82 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide TIP Click on Digitization Epi illumination in Preference in the Edit menu and select YES for Invert Pixel data ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi ilumination Timer Invert pixel data Auto Range ws Ono 5 4 Program settings Program Settings are used to set exposure time and exposure intervals when Program mode is selected as Exposure Type The input settings are saved as a template file Program Exposure Setting 1 Template ___ NewTempffte 7 Sensitivity Template Comment Exposure Count L Image 1 Image 2 Image 3 Image 4 Wait 0 j Wait Wait Wait Exposure Exposure Exposure Exposure Image 5 Image 6 Image 7 Image 3 Wait Wait Wait Wait Exposure j Exposure Exposure S Exposure Image 9 Image 10 Image 11 Image 12 Wait Wait L Wait L Wait
35. when you click the Start button ImageQuant LAS 4000 User Manual 28 9607 42 AB 85 86 5 Software reference guide 5 6 Start During exposure type Precision During exposure a progress bar and ending time are displayed to indicate the progress status of exposure ImageQuant LAS 4000 File Edit View Option Help ImageQuant LAS 4000 Exposure Type Precision Y Exposure Time Auto Manual 30 sec Y Sensitivity Resolution High Resolution b d Exposure Time 30sec Finishing Time AM 09 40 Exposing Mananami Fluorescence EtBr Flat frame Auto Click Cancel to discontinue exposure The current image is not saved ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 6 Start During exposure type Increment or Repetition Ti 1 2 3 4 5 8 6 No Description Description 1 Range Scope can be changed by dragging 5 The changed gradation is applied to all im the mouse ages 2 Gradation can be changed by dragging the 6 Exposure is discontinued mouse The current image is discarded 3 Gradation conversion curve can be changed 7 Click this tab to enter index view linear or sigmoidal 4 Returns the gradation to the initial setting 8 Click this tab to magnify the selected image TIP e Exposure is interrupted when you click the Stop button during Precision exposure In this case the image in course of exposure is
36. 0 Edit menu Cut Cuts the selected characters This function can be used during manual input of exposure time and interval Copy Copies the selected characters This function can be used during manual input of exposure time and interval Paste Pastes the copied characters This function can be used during manual input of exposure time and interval ImageQuant LAS 4000 User Manual 28 9607 42 AB 95 5 Software reference guide 5 10 Edit menu Preference The available settings can be viewed by clicking the respective item CCD Cooling Sets the CCD temperature ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer CCD Cooling Temperature Auto Range Cancel Root Folder Sets the folder in which files are saved ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer Auto Range C Documents and Settings TE ts ImageQuant LAS 4000 Image 96 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 10 Edit menu File Format Select the default format for saving images ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer
37. 00 Confirm the sample position and focus Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Precision for Exposure Type Exposure Type 3 Fv Precision Increment Epost reem ma Program ImageQuant LAS 4000 User Manual 28 9607 42 AB 53 4 Operating the ImageQuant LAS 4000 4 8 Exposing dye stained samples and films White Epi light Step Action Select Auto or Manual for Exposure Time Auto Manual STT Select an exposure time from the drop down list or enter the exposure time manually The exposure time can be Ex Ti PO ato a set from 0 01 seconds up to 30 hours Manual Exposure Type Note Automatic exposure o e time setting may Sine 4 not be possible de E Sensitivity pending on the su Add Digitiz 19 sample type and Ez method ae 30min 10min Smin 2min Amin Isec 12sec 14sec Flatframe 18sec Auto UtSsec 1 30sec 1 60sec 1 100sec Select Sensitivity Resolution Sensitivity Resolution LJ High Resolution Standard High Super Ultra High Binning Super Binning Ultra Binning TIP Select Sensitivity Resolution in the Help menu You can display Help that de scribes the relation between sensitivity and resolution 5 SensitivityResolution 6 Click the Start button The orange Busy LED lights on the instrument and a progress d
38. 575DF20 0 85 Epi Cy3 Fluorescence Cy3 Green 520nm 575DF20 0 85 Epi Cy5 Fluorescence Cy5 Red 630nm R670 0 85 Epi Alexa 633 Fluorescence Cy5 Red 630nm R670 0 85 Epi Alexa 660 Fluorescence Cy5 Red 630nm R670 0 85 Epi 126 ImageQuant LAS 4000 User Manual 28 9607 42 AB Classification Reagent name A Appendix A 3 Detection Reagents and corresponding settings ImageQuant LAS 4000 setting Method Light Filter 1 Attophos cannot be used for detecting nucleic acid on a nylon membrane Note Alexa 680 Fluorescence Cy5 Red 630nm R670 0 85 Epi Q dot Fluorescence DAPI UV 365nm Epi L41 2 8 Dy781 Fluorescence IR Infrared IR785 0 85 710nm Epi Alexa 680 Fluorescence IR Infrared IR785 0 85 710nm Epi Alexa 700 Fluorescence IR Infrared IR785 0 85 710nm Epi Attophos Fluorescence SYBRGreen Blue 460nm Y515 Di 0 85 Fluorescence dye Chemi Epi fluorescence ECL Plus Fluorescence SYBRGreen Blue 460nm Y515 Di 0 85 Epi Silver stain Digitization Trans White Trans Through 2 8 CBB stain Digitization Trans White Trans Through 2 8 Digitization X ray film Digitization Trans White Trans Through 2 8 NBT BCIP Digitization Epi White Epi Through 28 With regard to patents owned by third parties related to among other things sample preparation we recommend that you consult with a lawyer or patent attorney about obtaining a license fro
39. 6 Start Description Description 1 The range can be changed by dragging the Print image or screen mouse 2 Gradation can be changed by dragging the Save image mouse 3 The gradation conversion curve can be Return to main screen changed linear or sigmoidal ImageQuant LAS 4000 User Manual 28 9607 42 AB 89 5 Software reference guide 5 6 Start After exposure type Increment Repetition or Program For magnified image view For index image view No Description No Description 1 The range can be changed by dragging the 6 Return to main screen mouse 2 Gradation can be changed by dragging the 7 Save image mouse 3 The gradation conversion curve can be 8 Print image or screen changed linear or sigmoidal 4 The changed gradation is applied to allim 9 The selected image is marked by a yellow ages frame Click on an image to select it Double click an image or click the Image tab to open the magnified image view 5 Returns the gradation to the initial setting 5 7 Sensitivity Resolution This function sets the sensitivity and resolution 90 ImageQuant LAS 4000 User Manual 28 9607 42 AB The relation between the sensitivity and resolution is shown below SonsitivityResolation File size for each sensitivity 5 Software reference guide 5 7 Sensitivity Resolution High Resolution 12 6MB Standard 3 15MB High 3 15MB Super 3 15MB Ultra 3 15MB H
40. 84 images No Description No Description 1 Save Selected Image 4 Save Images in temporary folder 4 Exposure Type Increment Repetition Max84 Program Image files not displayed in the Index For multiple files specify the range of view are saved in the 100 latest im files to be saved ages maximum 84 files 2 Save Display image Max16 5 Comment Exposure Type Increment Repetition When the file is saved in gel format Program gel a comment can be entered in All image files displayed in the index the file view are saved maximum 16 files When the file is saved in TIFF format the comment will not be saved 3 Make Increment Image Exposure Type Program only Files of multiple exposures are accu mulated and saved The range of image files to be pro cessed in accumulation can be speci fied ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 13 Saving exposed images Step Action The Save dialog is displayed Type in a File name and press the Save button to save the file LAS 4000Image a 1 My Recent Documents Desktop My Network File name ae 2 3 Places Save as type Gel File Format gel 3 No Description 1 Select where to save the file 2 The file name is set automatically but may be replaced File names for single and programmed exposures are automatically set in the format lt File
41. AS 4000 User Manual 28 9607 42 AB 107 5 Software reference guide 5 13 Help menu About ImageQuant LAS 4000 Control Software The version information of Image Reader is displayed Click to close the window ImageQuant LAS 4000 iMagination ot work Version 1 1 X Build 1 1 0 17 SiN 9678 GONTROL SOFTWARE Instrument S N 9678001 9 200 GENERAL ELECTRIC COMPANY ALL RIGHTS RESERVED Technical support http y wwuw gelifesciences com cor 108 ImageQuant LAS 4000 User Manual 28 9607 42 AB 6 Installing ImageQuant LAS 4000 Control Software 6 Installing ImageQuant LAS 4000 Control Software Administrator priveleges required All tasks related to software installation or uninstallation require a computer account with administrator priveleges 6 1 Installation sequence Software installation is performed in the following sequence 1 Install the USB contol driver 2 Install the USB function driver 3 Install the ImageQuant LAS 4000 Control Software 6 2 Install ImageQuant LAS 4000 Control Software under Windows XP Before you begin Log in using a Windows account with administrator privileges Install the USB Control Driver Windows XP Step Action il Disconnect ImageQuant LAS 4000 from the computer 2 Open the control panel and select Printers and Other Hardware 3 Click Add Hardware 4 Click the Next button in the Add hardware wizard 5 Select Yes have already connected the hardware and cl
42. Camera cable USB cable Note 12 Do not connect cables other than the camera cable supplied with the equipment ImageQuant LAS 4000 User Manual 28 9607 42 AB 2 The ImageQuant LAS 4000 2 3 Connections Note Do not disconnect the camera cable other than when removing the camera head LAS High Sens lens High Sens lens cable A cable used for connecting the LAS High Sens lens and the IDX Make sure that the cable is securely connected Note e Never remove the cable except when replacing the lens e Be sure to turn off the equipment before replacing the lens Epi lights Connector on Epi light source The Epi light sources illuminate the object to be imaged from the upper right and upper left The connector is connected to the Epi light source connector inside ee the IDX Figure 2 1 Incident Epi light source connector rear Epi light source connector A connector used for connecting the Epi light source Power is supplied to the Epi light source through this connector Figure 2 2 Inside the instrument Note e The Epilight sources for the right and left sides are different Attach the appropriate source on respective sides e The connector will be connected when the Epi light source is inserted Make sure that the connector is securely connected See Section 3 3 2 Changing the Epi lights on page 29 e Be sure to use Epi light sources of the same type for the right and
43. Epi illumination tab in Edit Pref erence and make sure that YES is se lected for Invert Pixel data If this is set to NO the data will not be inverted and the quantitative value will be re versed ImageQuant LAS 4000 User Manual 28 9607 42 AB 51 4 Operating the ImageQuant LAS 4000 4 8 Exposing dye stained samples and films White Epi light 3 ImageQuant LAS 4000 Fie Edt View Option Help ImageQuant LAS 4000 Exposure Type 3 L reason v Exposure Time 4 O Auto Manual isec Y 5 Sensitivity Resolution Standard Y Add Digitization image O on Flat frame Auto Bo i m O Step Action Click the Method Tray position button Result The Method Tray position dialog opens Method Tray position Method Fiter iis Chemiluminescence Through Fluorescence cy3 Y Green Y 575DF20 O Digitization Epi illumination White Epi illumination Through O Trans illumination White Trans illumination Through 1 144 x 96 mm ms j OK Cancel e Select Digitization and Epi illumination for Method e Select Tray position according to the sample used e Click the OK button 52 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 8 Exposing dye stained samples and films White Epi light Step Action Click the Focusing button Result The focusing controls appear Fie Ed Option Heip Image Quant LAS 40
44. GE Healthcare ImageQuant LAS 4000 User Manual Table of Contents L WAN UN sercsernenne e R 1 1 Important user iINfOrmation ou eeessessssssscssesststeesesceesssssnnnseceeseeceeesnuntseseccessessnnnnessseeeeees 2 ThelmageQuant LAS 4000 soars ace eri ee 20 The ImageQuant LAS 4000 Grtenerc c cccvesiccenneienimaraneienanaiouniens 2 2 Inside the ImageQuant LAS 4000 saccciccccsssessascsencocscdbensvas stoeneencosiassnenseentdccsnesdsavaebeeveonnee 23 COMI CHS nere E armani nian eon E EEEE BOR Pans PIG CCS Se ONCE eeann a REE E RERS 3 Exchanging accessory DOVES societies ceiniecctnanenannaniienemicees 3 1 Changing or installing a filter ccssecttsssseseessceerecstemtonrreieesnnsememmanennanonneineeienaates 3 2 Installing or exchanging the lens s ssssss1151100151100s015s00so1ss0rsorsrstrntsnornntssorsntsn nornos 3 2 1 Removing the LAS High Sens lens cscs cocacncacnaidannmoneaemnanas 3 2 2 Installing the LAS High Sens lens cnet isctmsemrnscnnananenisehenmimnannaene 3 2 3 Removing the wide view lens u eeessssssssssssscessssssssnesssessescessnnsetitsesseesessnsnneeeeseeeeeeesnnneees 3 24 Installing the wide View lenser eia a aa 3 3 Changing the light BOUNCES innnan 3 3 1 Changing the UV transilluminator or white light table w eesssecssssssssssssessssseessen 33 2 Changing the Epi l9NtS sisisssrisssisisssesisrssrisistissisens isst 4 Operating the ImageQuant LAS 4000 seesssssesssssssesssssresesssssssessssssessssseses
45. Latch Epi light base 1 If the tray is at Tray position 4 proceed to step 3 Otherwise close the IDX door 2 Click on Method Tray position set Tray position to 4 and click the OK button 3 Open the IDX door If no Epi light is currently in place inside the instrument proceed to step 7 ImageQuant LAS 4000 User Manual 28 9607 42 AB 29 3 Exchanging accessory parts 3 3 Changing the light sources 3 3 2 Changing the Epi lights 4 Spring up the latch to unlock the Epi light 7 Before installing the new Epi light ensure that the latch is sprung open upwards 30 ImageQuant LAS 4000 User Manual 28 9607 42 AB 3 Exchanging accessory parts 3 3 Changing the light sources 3 3 2 Changing the Epi lights 8 Hook the hanger located on top of the Epi light to the top edge of the Epi light base 1 2 No Action 1 Two guide holes are located on the back of the Epi light base 2 Slide the hook after attaching the guide pins to the guide holes 9 Push the Epi light while holding the latch open and slide it inwards until a click sound is heard 11 Repeat this process for the opposite side Note e The Epi lights are clearly labelled L for the left and R for the right sides e Install the Epi lights in matching pairs only If Epi lights with different wavelengths are used the intended image will not be obtained ImageQuant LAS 4000 User Manual 28 9607 42 AB SL 4 Operating the ImageQuant LAS
46. Name gt _n where n is the next available index For images taken in exposure type Increment file names are automatically set in the format lt FileName gt _n_N_ lt TotalTime gt where n is the next available index N the total number of exposures and TotalTime is the total exposure time for the image 3 Select a file format The gel format is a GE Healthcare format that records details of the exposure Such details are not saved in tif format TIP To automatically launch an external application to view images upon saving refer to Launch Applicaton on page 98 Note Special characters such as or other characters that have a special meaning in Windows should not be used in a file name 4 14 Ending the session This section describes how to end the current session with the ImageQuant LAS 4000 The instrument should be switched off as described below when it will not be used for some time 72 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 14 Ending the session Step Operation Select Quit from the File menu I ImageQuant LAS 4000 em Edit view Option Help Select Stop the CCD cooling now and click OK Stop the CCD cooling now O Keep the CCD cooling after quit TIP When Keep the CCD cooling after quit is selected the cooling temperature of the CCD will be maintained and the instrument can be used straight away the next time the ImageQuant
47. Y 605DF40 Digitization Epi illumination White Epi illumination Through Trans illumination White Trans illumination Through Tray position 105 x 70mm Za OK Cancel e Select Fluorescence as the Method and choose EtBr from the pull down menu The 605DF40 filter is automatically selected e Select the Tray position e Click the OK button ImageQuant LAS 4000 User Manual 28 9607 42 AB 43 4 Operating the ImageQuant LAS 4000 4 6 Exposing fluorescent samples EtBr Step Action Click the Focusing button Result The focusing controls appear 2 J w Brightness i E vl ii Adjust i Confirm the sample position and focus Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Precision for Exposure Type 3 Je Precision Be Program 44 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 6 Exposing fluorescent samples EtBr Step Action Select Auto or Manual for Exposure Time Auto Manual eae Select an exposure time from the drop down list or enter the exposure time manually The exposure time can be ee aaa S set from 0 01 seconds up to 30 hours Exposure Type Note Automatic exposure a time setting may aed 4 not be possible de pending on the sample type and method 1min 12sec 14sec Flat frame 18sec Auto 1 15sec 1 30
48. ally selected e Select the Tray position e Click the OK button 48 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 7 Exposing fluorescence Epi illumination Step Action Click the Focusing button Result the focusing controls appear eh sant LAS 4000 Fie EGR Vew Opin Help ImageQuant LAS 4000 Exposure Type Interval Time Maral a Sensitivity Resolution Repeat Limit Set Lette 100 times 1 100 2 Fluorescence cys Flat frame Confirm the sample position and focus Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Precision for Exposure Type Exposure Poemen O Auto Repetition Manu Program aaa ImageQuant LAS 4000 User Manual 28 9607 42 AB 49 4 Operating the ImageQuant LAS 4000 4 7 Exposing fluorescence Epi illumination Step Action Select Auto or Manual for Exposure Time Auto Manual STT Select an exposure time from the drop down list or enter the exposure time manually The exposure time can be Ex Ti PO ato a set from 0 01 seconds up to 30 hours Manual Exposure Type Note Automatic exposure ne ee time setting may Sine 4 not be possible de E Sensitivity pending on the su Add Digitiz 19 sample type and Ez method ae 30min 10min Smin 2min Amin Isec 12sec 14sec Flatframe 18sec Auto UtSsec
49. ane samples or films It is used together with the ImageQuant LAS 4000 Control Software to e Expose chemiluminescent samples or membranes e Expose fluorescent samples using epi incident or trans transmitted light sources for example gels stained by ethidium bromide EtBr The ImageQuant LAS 4000 can be supplied with NIR Red Green Blue and UV epi lights e Expose dye stained gels and membranes or films using epi or trans illumination e Expose a sample repeatedly incrementally or by programming exposure schemes Purpose of this manual The ImageQuant LAS 4000 User Manual provides detailed instructions on using the ImageQuant LAS 4000 and outlines the functions of the ImageQuant LAS 4000 Control Software Note Be sure to refer also to the manual Getting Started with ImageQuant LAS 4000 for regulatory information specifications and the troubleshooting guide 1 1 Important user information Read this before using the ImageQuant LAS 4000 AR All users must read the safety instructions in Getting started with ImageQuant LAS 4000 before installing using or maintaining the equipment Do not operate the ImageQuant LAS 4000 in any other way than described in the user documentation Otherwise you may be exposed to hazards that can lead to personal injury and you may cause damage to the equipment ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 1 Introduction 1 1 Important user information Intended use The Imag
50. ation 4 7 Exposing fluorescence Epi illumination This section describes how to expose fluorescent samples using Epi illumination Ensure that the correct Epi lights and filter are installed See Chapter 3 Exchanging accessory parts on page 17 Place a sample on the Epi tray Put the tray into the ImageQuant LAS 4000 and close the door Note e Make sure that sample is suitably placed on the tray for the desired tray position e Ensure that a suitable method exists and that the ImageQuant LAS 4000 is properly calibrated See Section 4 16 Creating a new method and performing flat frame calibration on page 78 ImageQuant LAS 4000 File Edit View Option Help ImageQuant LAS 4000 Add Digitization image O on Flat frame Auto isos ImageQuant LAS 4000 User Manual 28 9607 42 AB 47 4 Operating the ImageQuant LAS 4000 4 7 Exposing fluorescence Epi illumination Step Action Click the Method Tray position button Result The Method Tray position dialog opens Method Tray position Method Chemiluminescence Fluorescence cy3 v Green Epi RGB Vv 575DF20 O Diaitization Epi illumination White Epi illumination O Trans illumination White Trans illumination Tray position i OK Cancel e Select Fluorescence as the Method choose the appropriate fluorescent marker and the ap propriate light source from the pull down menus The appropriate filter is automatic
51. ct a method from the drop down menu at the top Put the displayed tray and Calibration Plate in the instrument Select the condition to be created Click the Start button Note Make sure that an appropriate filter and tray are in place nan FF WN N 6 Amessage window opens when the calibration is completed Click OK 7 Click the Close button to close the FlatFrame Calibration window ImageQuant LAS 4000 User Manual 28 9607 42 AB 105 5 Software reference guide 5 12 Option menu 5 12 2 Flat Frame Calibration Flat Frame Calibration Pro 1 If FlatFrame Calibration Pro is selected in the Option menu the window of the conditions preset in the Method Tray position window will be displayed FlatFrame Calibration Pro Light Filter is Tray Position White Epi White Through Lens FUJINON VRF43LMD ExposureTime O Auto Manual File Name Put the appropriate tray and calibration plate in the instrument Set the exposure time Enter a file name Click the Start button nan FF WN N 5 12 3 Method Setting Method Setting is used to set the content displayed on the Method Tray position screen bd Method Setting Method List Name Light Filter Cy2 RGB E Blue Epi RGB vsisoi Foss Cy2 E Blue Epi Blue vsispi Foes Cy3 RGB E creen Epi rGB 5750F20 Fo 85 Cy3 E Green Epi creen 5750F20 Fo as Cy5 RGB E Red Epi RGB prso Foss Cy5 E Red Epi Red R670 F0 85
52. d Tray position button Result The Method Tray position dialog is opened Method Tray position Method Chemiluminescence Fluorescence tcy z Ys150 Digitization Epi illumination white Epi illumination Trans illumination White Trans illumination 105 x 70mm L LO OK Cancel Select the Method and Tray Position Click the OK button ImageQuant LAS 4000 User Manual 28 9607 42 AB 65 4 Operating the ImageQuant LAS 4000 4 11 Exposing repeatedly Repetition Step Action Click the Focusing button Result The focusing controls appear eh ant LAS 4000 Fie EGR Vew Opon Help ImageQuant LAS 4000 Exposure Type Interval Time Manual Sensitivity Resolution Repeat Limit Set Lene 100 times 1 100 2 READY F Nsorescence cya Flat frame CK brightness ED HE Aut O LJ VID a Confirm the sample position and focus Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Repetition for Exposure Type Exposure Type 3 Re Precision Interval Tin crement v Repetition Manu Program o Select Interval Time unit and enter a numeric value Interval Time LONER to O w 4 Sensitivity Resi sondar ME Repeat Limit Set Limit Note Interval Time can be entered in the range of 10 seconds to 2 hours 66 ImageQuant LAS 4000 User Manua
53. d in the range of 10 seconds to 2 hours ImageQuant LAS 4000 User Manual 28 9607 42 AB 61 4 Operating the ImageQuant LAS 4000 4 10 Exposing consecutively Increment Step Action Select Sensitivity Resolution Binni Super Binning Ultra Binning TIP Select Sensitivity Resolution in the Help menu You can display Help that de scribes the relation between sensitivity and resolution 5 To specify the maximum number of exposures to be taken check the Set Limit check box and specify the number of images 6 Popart mmi 100 times 1 100 62 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 10 Exposing consecutively Increment Step Action Click the Start button to begin exposure The index view and expanded view can be switched by clicking the Index or Image tab Note The displayed expanded image is the latest image Index view Image view m i Coe ie mogeQuan Tere Al Exposure automatically stops after the number of images set in Repeat Limit Note Clicking the Stop button cancels the current exposure TIP It is possible to stop exposures automatically when the image is saturated This function is activated in Edit Preference under the Increment tab TIP During exposure the gradation conversion curve can be changed and the grada tion adjusted When the Apply All button is clicked the adjusted gradation will
54. direct sunlight Attach a blind to nearby windows 5 Donotplace objects near the power outlet so that you can disconnect the power cable from the outlet in case of emergency Floor oscillation conditions 1 Operating time Oscillation Impact 2 Non operating time Oscillation Impact 0 03G 5 to 60 Hz 1G 0 4G 5 to 60 Hz 2G ImageQuant LAS 4000 User Manual 28 9607 42 AB A Appendix A 4 Main specifications Environmental conditions Operating temperature humidity conditions Temperature 15 C to 28 C with temperature fluctuation below 10 C per hour or lower Humidity 30 to 70 RH_ no dew condensation When the above conditions cannot be satisfied modify the facilities accordingly Transportation storage conditions Temperature 25 C to 70 C Humidity 5 to 100 RH no dew condensation Installation location conditions 1 Do not install the equipment in an area where the temperature varies widely Do not install the equipment near a source of heat Do not install the equipment in an area where it may get wet or flooded Do not install the equipment in an area where it may be exposed to corrosive gas Do not install the equipment in a dusty area Dn nO FF WN N Do not install the equipment in an area constantly or excessively exposed to os cillations or impacts 7 Donot install the equipment in an area exposed to direct sunlight Operation site Indoors Maximum operating alti
55. e inf X Files of type Setup Information inf ImageQuant LAS 4000 User Manual 28 9607 42 AB Step 6 Installing ImageQuant LAS 4000 Control Software 6 3 Install ImageQuant LAS 4000 Control Software under Windows Vista Action 10 11 1e 15 14 15 16 Select the USB Control folder and click Open Lookin DVD CD RW Drive D ImageQuant LAS 40 P O Ey P Name Size Type Date modif Location a ay Files Currently on the Disc 2 a Recent Places pres i USB Control m i File Folder oS cern en en en see Desktop l admin Computer L lt Network USB Function File Folder File name int X Files of type Setup Information inf Select the file DevMng and click the Open button Lockey fy USE Contest OF 7a E Nare Oete most Type See gt Qos Recent Places Destop wn L Computer Network teame Oer g Fidea sien C Trea Click the OK button in the dialog Install from disk Click the Next button in the wizard Add hardware Click the Next button once again The following warning is displayed Proceed by cliking Install this driver software anyway gt Don t install this driver software You should check your manutacturer s webute for updated drover software for your dece gt Install this driver software anyway Only install driver software obtained from your manufacturers website or disc Unsigned software from other sources may
56. eQuant LAS 4000 Control Software CD My Recent Docurerts Deikko O Shred Documents UD Mr Documents Sj My Nernoh Pisces My Documents iga My Computes Fie poe ri Open My Nebr Fien of pe Coce ImageQuant LAS 4000 User Manual 28 9607 42 AB Step 6 Installing ImageQuant LAS 4000 Control Software 6 2 Install ImageQuant LAS 4000 Control Software under Windows xP Action 12 15 14 15 16 17 18 Open the USB Control folder Locke i bnageQuart LAS 4000101 lt 08 co 4 QU heen My Racer Documents Daiko My Documents My Comptes Fie pane nt Open My Neima Fien of ype Concel Select the DevMng inf file and click the Open button Locate File Look in USB Control a Fiename DevMng x Files of ype Click the OK button in the Install from disk dialog Click the Next button in the Add hardware wizard Click the Next button again Click the Continue Anyway button in the Hardware Installation dialog Hardware Installation The software you are installing for this hardware FLA LAS USB Device Managment Driver has not passed Windows Logo testing to verify its compatibility with Windows XP Tell me why this testing is important Continuing your installation of this software may impair or destabilize the correct operation of your system either immediately or in the future Microsoft strongly recommends that you stop this installation now and c
57. eQuant LAS 4000 is a camera system that produces digital images of chemiluminescent dyed or fluorescent gels and membranes The ImageQuant LAS 4000 is intended for research use only and shall not be used in any clinical procedures or for diagnostic purposes Safety notices The user documentation contains WARNINGS CAUTIONS and NOTICES concerning the safe use of the product See definitions below Warnings WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is important not to proceed until all stated conditions are met and clearly understood Cautions CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to proceed until all stated conditions are met and clearly understood Notices NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment Regulations and standards supplementary information This equipment conforms to the regulations and standards described below EMC VCCI Class A Conformance FCC Part 15B Class A ICES 003 Class A IEC 61326 1 2005 EN 61326 1 2006 6 ImageQuant LAS 4000 User Manual 28 9607 42 AB 1 Introduction 1 1 Important user information Safety UL61010 1 second edition CAN CSA C22 2 No 61010 1 second edition IEC61010 1 2001 second edition IEC61010 2 081 f
58. ect that radiates light at the same brightness is represented evenly in any section of an exposure area A correction file for correcting unevenness is called a flat frame Under the same conditions tray position iris fluorescent filter light source and tray as when exposing an object sample a flat frame records the image in a uniform fluorescent plate fluorescent mode or white plate digitize mode ImageQuant LAS 4000 User Manual 28 9607 42 AB 121 A Appendix A 1 Glossary Conceptual diagram of CCD calibration and flat frame Virtual profile in which light is supposed to be uniformly emitted from the whole image Electrical noise component Optical unevenness Sample image Center CCD Calibration Data Center FlatFrame Center A 2 Quick reference functions Raw image State of image that is not corrected Center 4 DX Raw image CCD Calibration Data Removal of noise component Center 4 DX Raw image CCD Calibration Data FlatFrame Limb darkening due to optical characteristics Center Function Function name Set item Exposing for a fixed exposure Exposure Type Precision time Exposing consecutively Exposure Type Increment Exposing repeatedly Exposure Type Repetition 122 ImageQuant LAS 4000 User Manual 28 9607 42 AB ImageQuant LAS 4000 A Appendix A 2 Quick reference functions Function Function name Set item Dete
59. elect Digitization Trans illumination for Method 2 Select Tray position according to the sample used 3 Click the OK button 56 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 9 Exposing dye stained samples and films Trans illumination Step Action Click the Focusing button Result The focusing controls appear 3 ImageQuant LAS 4000 Fle Edt View Option Help ImageQuant 2 Je Brightness j 7 a Adjust L o a Confirm the sample position and the focus Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Precision for Exposure Type Exposure Type 3 v Precision Increment Repetition Program ImageQuant LAS 4000 User Manual 28 9607 42 AB 57 4 Operating the ImageQuant LAS 4000 4 9 Exposing dye stained samples and films Trans illumination Step Action Select Auto or Manual for Exposure Time Auto Manual EN Select an exposure time from the drop down list or enter the exposure time manually The exposure time can be set Ex Ti PO ato ime from 0 01 seconds up to 30 hours Manual Exposure Type Note Automatic exposure time setting may not Sine 4 be possible depend m n nsitivity ing on the sample su mee Shr type and method Add Digitiz ax Oo 60min 30min 10min Smin 2min Amin sec 12sec 14sec Flat frame 1 8sec
60. ep Action Adjust the gradations of the exposed image then save and print the image ImageQuant LAS 4000 Fie EGR Vew Option Hep ImageQuant LAS 4000 Exposure Type E Complete _ Click the Complete button Result The display returns to the main screen 4 6 Exposing fluorescent samples EtBr This section describes how to expose ethidium bromide EtBr stained samples using the UV transilluminator 312 nm Place a gel sheet which has been cut out to be slightly larger than the sample onto the UV trans tray Place the sample on the gel sheet Put the tray into the ImageQuant LAS 4000 and close the door Note The 605DF40 filter is normally used If this filter is currently not in the filter turret it should be installed Refer to Section 3 1 Changing or installing a filter on page 17 Note Ensure that the UV transilluminator is installed For instructions on changing the trans light source see Section 3 3 1 Changing the UV transilluminator or white light table on page 28 42 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 6 Exposing fluorescent samples EtBr IF ImageQuant LAS 4000 Ea View Opon Help ImageQuant 3 4 5 Sensitiv ieoi z 1 2 6 Step Action Click the Method Tray position button Result The Method Tray position dialog opens Method Tray position Method Chemiluminescence Through Fluorescence UY Trans illumination W
61. harm your computer or steal information vi See dette Click the Finish button in the Add Hardware wizard to complete the installation ImageQuant LAS 4000 User Manual 28 9607 42 AB 115 6 Installing ImageQuant LAS 4000 Control Software 6 3 Install ImageQuant LAS 4000 Control Software under Windows Vista Install the USB function driver Windows Vista Note During software installations you may be asked to confirm your actions in a dialog with the text Windows needs your permission to continue Enter an administrator password if prompted then click Continue to proceed with the installation Step Action 1 Connect the computer and the ImageQuant LAS 4000 with a USB cable and turn ON the power switch of ImageQuant LAS 4000 Result The scanner will automatically be detected by the computer and the Plug and Play function in Windows Vista starts 2 In the Found New Hardware dialog select Locate and install driver software recom mended 3 Insert the ImageQuant LAS 4000 Control Software CD and click the Next button in the Found New Hardware dialog 4 Select Install this driver software anyway v See dette 5 A successful installation message appears Click the Close button Install ImageQuant LAS 4000 Control Software Windows Vista 116 Note During software installations you may be asked to confirm your actions in a dialog with the text Windows needs your permission to continue Enter an admini
62. he light sources 3 3 1 Changing the UV transilluminator or white light table Type Name Peak wavelength UV Epi light 365 nm Blue Epi light 460 nm Green Epi light 520 nm Red Epi light 630 nm Incident RGB module 3 color Epi light 460nm or 520nm or 630 nm NIR Epi light 710 nm White Epi light 5 3 3 1 Changing the UV transilluminator or white light table The UV transilluminator and white light table are exchanged as follows 1 If neither the UV transilluminator or white light table are currently in place inside the instrument proceed to step 4 2 Turn the lever holding the light source to unlock it 4 er a 3 Pull out the light source carefully along the guide while holding it with both hands 28 ImageQuant LAS 4000 User Manual 28 9607 42 AB 3 Exchanging accessory parts 3 3 Changing the light sources 3 3 1 Changing the UV transilluminator or white light table 4 Push the desired light source in along the guide at the bottom of the IDX 3 3 2 Changing the Epi lights This section describes how to change a set of Epi lights The RGB module Epi lights are also installed as described here Note The tray should be at tray position 4 before installing Epi lights If the tray is higher then it should be lowered to tray position 4 using the ImageQuant LAS 4000 Control Software before attaching the lights as described below
63. ialog is displayed during exposure 54 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 8 Exposing dye stained samples and films White Epi light Step Action Adjust the gradations of the exposed image then save and print the image Click the Complete button The display returns to the main screen 4 9 Exposing dye stained samples and films Trans illumination This section describes how to expose dye stained samples using white trans illumination Place a sample on the White trans tray Put the tray in the IDX and close the door ImageQuant LAS 4000 User Manual 28 9607 42 AB 55 4 Operating the ImageQuant LAS 4000 4 9 Exposing dye stained samples and films Trans illumination ImageQuant LAS 4000 Fie Edt View Option Help ImageQuant LAS 4000 Exposure Type 3 L reson v Exposure Time 4 O Auto Manual isec Y 5 Sensitivity Resolution Standard Y Add Digitization image O on Flat frame Auto LL i as Se Step Action Click the Method Tray position button Result The Method Tray position dialog opens Method Tray position Method F a O Chemiluminescence Through Fluorescence EtBr Y UY Trans illumination Y 605DF40 Digitization Epi illumination White Epi illumination Through Trans illumination White Trans illumination Through 1 105 x 70mm L OK Cancel 1 S
64. ick the Next button ImageQuant LAS 4000 User Manual 28 9607 42 AB 109 110 6 Installing ImageQuant LAS 4000 Control Software 6 2 Install ImageQuant LAS 4000 Control Software under Windows XP Step Action 6 10 11 Select Add a new hardware device and click the Next button Add Hardware Wizard The following hardware is already installed on your computer ON S Fiom the list below select an installed hardware device then click Next to check properties or troubleshoot a problem you might be having To add hardware not shown in the list click Add a new hardware device Installed hardware USB Root Hub amp USB Mass Storage Device amp Genetic USB Hub amp USB Composite Device S Generic volume Add a new hardware device Newt gt Select Install the hardware that I manually select from a list Advanced and click the Next button Select Show All Devices and click the Next button From the fat below select the type of hardware you are installing 1I you do ne see the hardener calngery you wand click Show AI Dewees Common hardware typer Dusty adapters h Sen over GIDE TATA controters IEEE 1394 Bus hont conmoers P imaging devices Ay irked devices intel AM Codec E riere i eck Cie C cca Click the Have Disk button in the Add hardware wizard Insert the ImageQuant LAS 4000 Control Software CD and click the Browse button Select to install the driver from the Imag
65. igh Binning 786MB Super Binning 197MB Ultra Binning 49 2MB TIP Select Sensitivity Resolution in the Help menu The relation between the sensitivity and resolution is displayed 5 8 Method Tray position The window for setting the method and tray position is displayed when you click the Method Tray position button Select a method and tray position most suitable for the sample and click the OK button ImageQuant LAS 4000 User Manual 28 9607 42 AB 91 5 Software reference guide 5 8 Method Tray position Method Tray position Method Filter a Chemiluminescence Through Fluorescence jag v ris Digitization Epi illumination White Epi illumination Through Trans illumination White Trans illumination Through 105 x 70mm m OK Cancel Set the method and tray position Method The method the combination of the optimum light source filter and iris is registered for each detection method When a method is selected the light source filter and iris are set automatically The contents of each setting are shown in the table below Note The RGB module may be used instead of the separate red green and blue Epi lights Method Option Light Filter lris Chemiluminescence None Through FO 85 SYBR Green Blue 460nm Epi Y515 Di FO 85 GFP Blue 460nm Epi 510DF10 F0 85 EtBr UV 312nm Trans illumi 605DF40 F2 8 nation Fluorescence Cy2 Blue 460nm Epi Y515 Di
66. ired by the ImageQuant LAS 4000 Control Software They are not deleted during the uninstallation and remain in the Data folder of ImageQuant LAS 4000 Control Software folder Uninstalling ImageQuant LAS 4000 Control Software under Windows Vista Step Action 1 118 Open the control panel and Select Uninstall a program under Programs ImageQuant LAS 4000 User Manual 28 9607 42 AB 7 Uninstalling and upgrading ImageQuant LAS 4000 Control Software Step Action 2 Select ImageQuant LAS 4000 and then click Uninstall OOE meie ore orea iae 3 Confirm the uninstallation by clicking Yes when prompted 4 If User Account Control UAC is enabled in Windows Vista a dialog displays the message An unidentified program wants access to your computer Click Allow Upgrading ImageQuant LAS 4000 Control Software Step Action 1 Uninstall the current version of ImageQuant LAS 4000 Control Software 2 Install the new version of ImageQuant LAS 4000 Control Software ImageQuant LAS 4000 User Manual 28 9607 42 AB 119 8 After sales service 8 After sales service 8 1 Warranty 1 The warranty period will expire after 1 year from the date the system was delivered 2 GE Healthcare will make repairs free of charge for failures during the warranty period provided that normal usage conditions and the instructions given in this manual etc are followed 3 Repairs of the following failures will be charged for even
67. irictiansonciemtaeedeanienlacim S1 EMO SUS AYE sre E E E EE 5 2 EADS US ME siaiisisirdetsintacsinieninaasancuoinienananiaeimngoninanONne 5 3 EI digitization IMAQE nga scsxsscasccasazsescacpopcschsosouncesocoasoausnsessecunobassasasebensuscososuossvarscbeiesnnen Ba Progra SeAOS pnsmnncannncnanica nena NN TIN AERUERTE 55 FOCUSING sksini EE raar inno taea Ers ER a LEE Ea que TETEE EEEN a e UI E E E E E E EE ImageQuant LAS 4000 User Manual 28 9607 42 AB Table of Contents Table of Contents 5 7 Sengsitivity ResolUt ON eee eee ene nee ne RAS Pee ne oe eee meee oe 90 58 MEMS iray Positions erenneren ea A es ereerifeseenetee 91 S9 ol 121 e E AEE E E E EEE EEA E 94 es aE A ere S E E E E E E EE 95 5 AVEW IMEN a a A E E E E E E EES 103 512 OPINE in E A RAAEN 104 5121 CCD Ss 51502 iise E a E AR 104 5122 Flat Fram Cabra ON neesiet e ei S EE a E E i TE EE EEEN 105 5 12 3 Method Setting eck esc tnnessancatasi asset bints neato cash arabs capta aaa SR ao eeu ate oe mem 106 513 Help UVM VA tases eiceetesscescenistaescceastecteetiacad cesses eaa aen a E RN 107 6 Installing ImageQuant LAS 4000 Control Software sessssessssesessessssessesesssseseorssseresseseeesssssenessesessesssessese 109 6 1 I stalati n SegUENCEincienenensanaimienn nnna a ER ENERE 109 6 2 Install ImageQuant LAS 4000 Control Software under Windows XP sssessssssssssessssssssseesssssssuessesssneeeeseenss 109 6 3 Install ImageQuant LAS 4000 Control Software under Wind
68. irst edition EN 61010 1 2001 EN 61010 2 081 2002 Laser USA 21 CFR Chapter Subchapter J Part 1040 10 Laser Products IEC 60825 1 2001 EN 60825 1 1994 A11 1996 A2 2001 CE LV Directive 2006 95 EC as amended EMC Directive 2004 108 EC as amended Machinery Directive 2006 42 EC Note This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to Part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his own expense This Class A digital apparatus complies with Canadian ICES 003 Cet appareil num rique de la classe A conforme la norme NMB 003 du Canada Note This is a class A product In a domestic environment this product may cause radio interference in which case the user may be required to take adequate measures Information for traceability Manufacturer GE Healthcare Contact information See back cover Power cable 1 In Japan and North America Do not use cables other than the power cable that comes wi
69. l 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 11 Exposing repeatedly Repetition Step Action Select Sensitivity Resolution Sensitivity Resolution L v High Resolution Standard High Super Ultra High Binning Super Binning Ultra Binning TIP Select Sensitivity Resolution in the Help menu You can display Help that de scribes the relation between sensitivity and resolution 5 To specify the maximum number of exposures to be taken check the Set Limit check box and Z specify the number of images Repeat Limit Set Limit 100 times 1 100 ImageQuant LAS 4000 User Manual 28 9607 42 AB 67 4 Operating the ImageQuant LAS 4000 4 11 Exposing repeatedly Repetition Step Action Click the Start button to begin exposure The index view and expanded view can be switched by clicking the Index or Image tab Note The displayed expanded image is the latest image Index view Image view Far eset A REED compete Exposure automatically stops after the number of images set in Repeat Limit Note Clicking the Stop button cancels the current exposure TIP During exposure the gradation conversion curve can be changed and the grada tion adjusted When the Apply All button is clicked the adjusted gradation will be applied to all images NE i No Description No Description 1 Therangecanbechangedby 4 The changed gradation is ap
70. m contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com quantitative_imaging imagination at work GE imagination at work and GE monogram are trademarks of General Electric Company ImageQuant are trademarks of GE Healthcare companies All third party trademarks are the property of their respective owners 2009 General Electric Company All rights reserved First published Oct 2009 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare representative for the most current information GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 28 9607 42 AB 12 2009
71. m the third parties A 4 Main specifications Images can be continuously exposed for up to 30 hours using ImageQuant LAS 4000 Control Software CCD 3 200 000 pixels Fujifilm super CCD Cooling temperature 25 C Number of gradations Recorded image 16 bit Focusing 8 bit Exposure time 1 100 seconds to 2 hours ImageQuant LAS 4000 User Manual 28 9607 42 AB 127 A Appendix A 4 Main specifications Lens High sensitivity lens F0 85 43 mm Wide view lens F1 8 24 mm Shading correction Software system Maximum sample size 250 x 250 mm with F1 8 24 mm lens 210 x 140 mm with F0 85 43 mm lens Dynamic range 4 orders of magnitude Maximum image size 12 6 MB Light source Epi class 1 light source Blue Epi light 460 nm Green Epi light 520 nm Red Epi light 630 Nnm RGB Module Epi light 460 nm 520 nm or 630 nm NIR Epi light 710 nm Epi and Trans class 1M light source UV Epi light 365 nm UV transilluminator 312 nm Size 322 x 313 x 87 MM UV transmitted filter size 200 x 200 mm White transmitted light table White LED Size 322 x 313 x 87 mm External dimensions Camera head 224x 161x252 mm W H D IDX 510 x 730 x 480 mm W H D Weight Camera head 3 4 kg IDX 49 kg not including the light source lens and tray High sensitivity lens 4 5 kg 128 ImageQuant LAS 4000 User Manual 28 9607 42 AB A Appendix
72. n Lens Exchange LAS 4000 fem Help CCD Calibration FlatFrame Calibration FlatFrame Calibration Pro Filter Customization ad Lens Exchange Method Setting A message is displayed Lens Exchange Close the door and click OK ImageQuant LAS 4000 control software will shut down Do you want to continue Click OK Lens Exchange Switch off ImageQuant LAS 4000 instrument and change the lens Turn off the instrument using the power switch 4 Remove the end cap from the lens 5 Insert the lens along the lens guide with the red points facing towards you ImageQuant LAS 4000 User Manual 28 9607 42 AB 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 2 Installing the LAS High Sens lens Red points 7 Connect the lens cable Turn the knurling ring until it is securely fixed Knurling ring 7 Connector of the lens cable Note Be sure to connect the lens cable Otherwise the high sensitivity lens cannot be recognized This may cause malfunction of the equipment ImageQuant LAS 4000 User Manual 28 9607 42 AB 23 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 2 Installing the LAS High Sens lens 3 2 3 Removing the wide view lens 24 8 Remove the lens cap Lens cap 9 Install the lens hood ee Lens hood A 1 Close the instrument door If the power is turned off and the filter changer
73. n list or locate the application using the Select button e Check the Activate save and launch function checkbox Print Adjust Adjusts the print output size ImageQuant LAS 4000 Preference Digitization Epi illumination Timer Auto Range ImageQuant LAS 4000 User Manual 28 9607 42 AB 99 5 Software reference guide 5 10 Edit menu Increment ImageQuant LAS 4000 Preference Description YES During incremental imaging exposure is automatically stopped when the amount of satu rated data is too large NO During incremental imaging exposure is continued until the Stop button is pressed 100 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 10 Edit menu Digitization EPI ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Description YES Pixel data inversion is performed NO Pixel data inversion is not performed ImageQuant LAS 4000 User Manual 28 9607 42 AB 101 5 Software reference guide 5 10 Edit menu Timer ImageQuant LAS 4000 Preference CCD Cooling Root Folder File Format FlatFrame Mode Launch Application Print Adjust Increment Digitization Epi illumination Timer Cooling Stop Timer Auto Range 1hr v If the ImageQuant LAS 4000 is not used during the chosen time the cooling will be automatically shut off and the system will enter power saving mode Auto Range ImageQuant
74. ng the lens 3 2 4 Installing the wide view lens A message is displayed Lens Exchange Close the door and click OK ImageQuant LAS 4000 control software will shut down Do you want to continue Click OK 3 Lens Exchange Switch off ImageQuant LAS 4000 instrument and change the lens Turn off the instrument using the power switch 4 Remove the cap attached to the F mount adapter by turning it to the right F mount adapter 5 Remove the end cap from the wide view lens 6 Align the White point mark of the F mount adapter with the lens mark and turn to lock them together Hold the lens so that the red point is towards you F mount adapter Wide view lens 26 ImageQuant LAS 4000 User Manual 28 9607 42 AB 7 Fix the lens in position be turning it to the right 8 Remove the lens cap 9 Placel the lens hood on the lens 3 3 Changing the light sources 3 Exchanging accessory parts 3 2 Installing or exchanging the lens 3 2 4 Installing the wide view lens Both trans transmitted and epi incident light sources are available for exposure with the ImageQuant LAS 4000 This section describes how to change both types of light source The following table summarizes the properties of the various light sources Peak wavelength UV transilluminator Transmitted 312 nm White light table ImageQuant LAS 4000 User Manual 28 9607 42 AB 27 3 Exchanging accessory parts 3 3 Changing t
75. not saved e When you click the Apply All button during magnified image display the gradation changed in the expanded image display state is applied to all magnified images Note Only the 100 latest exposed images can be saved for exposure types Increment and Repetition ImageQuant LAS 4000 User Manual 28 9607 42 AB 87 5 Software reference guide 5 6 Start During exposure type Program 10 9 6 1 2 3 4 5 8 7 No Description Description 1 The range can be changed by dragging the 6 Exposure is discontinued MOUSE The current image is discarded 2 Gradation can be changed by dragging the 7 Click this tab to enter index view mouse 3 The gradation conversion curve can be 8 Click this tab to magnify the latest image changed linear or sigmoidal 4 Return to initial setting 9 Preset conditions 5 The changed gradation is applied to all im 10 The area where an image will be displayed ages TIP e Images for which exposure is completed are automatically saved ina file with name format lt FileName gt _ lt PhotographNo gt _ lt ExposureTime gt e Animage which is the accumulation of all exposed images is saved in a file with name format lt FileName gt _ lt Sum gt _ lt TotalExposureTime gt Note Only the 16 latest exposed images can be saved for exposure type Program 88 ImageQuant LAS 4000 User Manual 28 9607 42 AB After exposure type Precision 5 Software reference guide 5
76. nt LAS 4000 Control Software This will ensure that necessary folders and files are created ImageQuant LAS 4000 User Manual 28 9607 42 AB 117 7 Uninstalling and upgrading ImageQuant LAS 4000 Control Software 7 Uninstalling and upgrading ImageQuant LAS 4000 Control Software Before you begin Log in using a Windows account with administrator privileges Uninstalling ImageQuant LAS 4000 Control Software under Windows XP Step Action 1 2 Open the control panel and select Add or Remove Programs Select ImageQuant LAS 4000 and click the Remove button E Add or Remove Programs Access and Defaults Currently installed programs and updates B Intel R Graphics Mex E Intel R Management Engine Interface E9 Intel R Network Connections 13 1 33 0 E Intel Active Management Technology 4 Java TM 6 Update 7 fB Microsoft NET Framework 1 1 B Microsoft NET Framework 1 1 Hotfix KB928366 jB Microsoft NET Framework 2 0 Service Pack 2 J NET Framework PrexP gD KB958481 B Dr Watson 5 NET Framework CRT NET Framework CA BP NET Framework CLR B NET Framework WinForms Show updates Sort by Name 0 01MB 4 89MB 0 18MB 171 00MB 185 00MB Installed On 6 11 2009 Installed On 6 11 2009 Installed On 6 11 2009 Installed On 6 11 2009 Installed On 6 11 2009 Installed On 6 11 2009 Installed On 6 11 2009 a Note Correction files created during calibration are requ
77. ontact the hardware vendor for software that has passed Windows Logo testing Continue Anyway i STOP Installation Click the Finish button to complete the driver installation ImageQuant LAS 4000 User Manual 28 9607 42 AB 111 6 Installing ImageQuant LAS 4000 Control Software 6 2 Install ImageQuant LAS 4000 Control Software under Windows XP Install the USB function driver Windows XP Step Action 1 Connect the computer and the ImageQuant LAS 4000 with a USB cable and turn ON the power switch of ImageQuant LAS 4000 The scanner is automatically detected by the computer In the Found New Hardware Wizard dialog choose No not this time Found New Hardware Wizard Welcome to the Found New Hardware Wizard Click the Next button in the Found New Hardware Wizard dialog Insert the installation CD Select Install the software automatically Recommended Click the Next button in the Found New Hardware Wizard dialog Click the Finish button to complete the installation Install ImageQuant LAS 4000 Control Software Windows XP Step Action 1 2 112 Insert the ImageQuant LAS 4000 Control Software CD Locate and double click the file ImageQuant LAS 4000 msi In the ImageQuant LAS 4000 InstallShield Wizard click the Next button Read the license text If the license agreement is not acceptable please contact a GE Healthcare representative see back cover of this manual for contact information
78. optimal use of the product TIP A tip contains useful information that can improve or optimize your procedures ImageQuant LAS 4000 User Manual 28 9607 42 AB 1 Introduction 1 1 Important user information Typographical conventions Software items are identified in the text by bold italic text A colon separates menu levels thus File Open refers to the Open command in the File menu Hardware items are identified in the text by bold text e g Power switch ImageQuant LAS 4000 User Manual 28 9607 42 AB 9 2 The ImageQuant LAS 4000 2 The ImageQuant LAS 4000 This chapter describes the features and part names of the ImageQuant LAS 4000 hardware and the connectors that connect the various parts of the equipment 2 1 The ImageQuant LAS 4000 exterior 10 Part Name Description 1 Camera head CCD cooling and image data output 2 Indicator panel Status lights 3 Power LED Lights when the power is on 4 Busy LED Lights when an exposure is in progress 5 Error LED Lights when an error is detected 6 Power switch Power ON O Power OFF 7 Intelligent dark box IDX Dark box ImageQuant LAS 4000 User Manual 28 9607 42 AB 2 The ImageQuant LAS 4000 2 2 Inside the ImageQuant LAS 4000 2 2 Inside the ImageQuant LAS 4000 Part Name 1 Epi light connector Description Connects the epi light to the IDX 2 Epi light s
79. ource NIR Epi light 710 nm Red Epi light 630 nm Green Epi light 520 nm Blue Epi light 460 nm RGB Module Epi light with red green and blue LEDs UV Epi light 365 nm White Epi light 3 Sample tray Supports the sample 4 Lifting table Moves the sample to the selected tray position 5 Lens F0 85 43 mm LAS High Sens lens or F1 8 24 mm wide view lens 6 Lens connector Connects lens to IDX ImageQuant LAS 4000 User Manual 28 9607 42 AB 11 2 The ImageQuant LAS 4000 2 2 Inside the ImageQuant LAS 4000 Part 7 Name Automatic filter changer Description Used to hold filters selected in the control software The follow ing filters are commonly used IR785 Alexa filter for NIR LED R670 Cy5 filter for red LED 575DF20 Cy3 filter for green LED 605DF40 EtBr filter for detecting EtBr 510DF10 filter for detecting GFP Y515 filter for blue LED L41 UV filter for UV LED Trans light source connec tor Connects the UV transilluminator or White light table to IDX Trans light source 312 nm UV transilluminator White light table Note Note Note Do not use connectors other than those supplied for the Epi light sources Do not connect cables other than those supplied to the lens Do not connect anything other than the transmitted light sources to the transmitted light source connector 2 5 Connections Camera head
80. ows VISTO ssssssssssssssssssssssststsesseeeeeseeee 113 7 Uninstalling and upgrading ImageQuant LAS 4000 Control Software ssssssesssssssesssssesssssssessssseesses 118 amp After Sales SC ON cose ra cae aecac sc cece saree ae se cece cece eee eee ei ee 120 Bele WACUOUVTY ccsssscesssetezsszcaszeccasssazszetenasssuszecens tng seseba same saccens dacs exfeuassceaqacegasstets stapatscetsatagesnseesa AEE A 120 82 REPONS eco cette ered eins tase detec e mans ernie ives ot ert eee ems 120 Bi E EIE ccs cael E T A cased sescaceuhaned A E E E E E SA E E 121 AL GOSSA aerei A A E E ee 121 A 2 Quick reference FUNCTIONS oecceeeccccecsseescsseccsssescsseccessesssssscssssccsssucssssssssssesssssesssusssusssssssessusssssusesssessiusessesessiusessesessiusessiees 122 A 3 Detection Reagents and corresponding Settings cscsssssssssssceccsssssssssesecesescsensssssesececsecssssssassssesesesesenseseses 125 A4 Main Specifications sssini sirrinin isitin Ein astat ENa Taa EA EEs ASETTA EEEE SESETAN AE 127 A 5 Minimum computer requirements cs ssasssscscassatcicarmenvecsnasvecswiersvaritarniavecsenremancinentsleennatoarnianaidenonnainnaaneetcs 132 PD Service teport fox SCRE sisces es csiscsstssssisscveicccedvesadiedsecasessrsasassnaais ees ca ha ANAA NRN ARTANA 133 4 ImageQuant LAS 4000 User Manual 28 9607 42 AB 1 Introduction 1 Introduction The ImageQuant LAS 4000 The ImageQuant LAS 4000 is a camera system for producing digital images of gel or membr
81. quired for creation is dis played 5 Click on the buttons in this column to check the boxes for both correction files for specific resolutions 104 ImageQuant LAS 4000 User Manual 28 9607 42 AB 5 Software reference guide 5 12 Option menu 5 12 1 CCD Calibration Description Description 3 Check individual check boxes to generate specific correction files 2 Click the Start button to generate the file e The progress of file creation is shown on the progress bar e To stop file creation click the Stop button The file will not be saved 5 12 2 Flat Frame Calibration This function creates a correction file Flat Frame that is required for correcting the optical characteristics The Flat Frame file varies depending on Light Filter Iris and Tray Position Note Administrator priveleges are required to perform Flat Frame calibration Flat Frame Calibration 1 Select Flat Frame Calibration in the Option menu The window below is then displayed FlatFrame Calibration Lite Digitization Epi ilumination W Applicable Tray EPI Tray Uniformity Standard Calibration Plate DI Calibrate All Tray Position eady Sun May 31 18 15 22 2009 Tray Position 1 Ready Mon May 25 14 16 55 2009 Tray Position 2 Tray Position 3 Mon May 25 14 17 36 2009 Ready Tray Position 4 Mon May 25 14 18 16 2009 Total Time For Calibration 0 minutes Sele
82. ray position button Result The Method Tray position dialog opens Method Tray position Method Filter aay Chemiluminescence Through F0 85 Fluorescence L wa Y515 Di O Digitization Epiillumination White Epi illumination Trans illumination White Trans illumination 1 Tray position p 105x 70 mm Select Chemiluminescence for Method e Select Tray position according to the sample e Click the OK button 38 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 5 Exposing chemiluminescent samples Step Action Click the Focusing button Result The focusing controls appear Fie Option Help ImageQuant LAS 4000 Sensitivity Resolution ie S E Adjust TY E Check that the sample is correctly postioned and focus the image Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Precision for Exposure Type Exposure Type 3 Pi v Precision Ep Osuna Increment E O Auto Repetition Manu Program ae ImageQuant LAS 4000 User Manual 28 9607 42 AB 39 40 4 Operating the ImageQuant LAS 4000 4 5 Exposing chemiluminescent samples Step Action Auto Exposure Type Exposure Time Auto O Manual Note Automatic exposure time setting may 4 not be possible de pending on the sample type and method Select Auto or Manual for Exposure Time Manual
83. rmining exposure time Exposure Time Auto automatically Entering and determining ex Exposure Time Manual posure time Determining a desired expo Exposure Type Program sure time and expose Setting the exposure time and the exposure intervals Exposure Type Program settings Setting sensitivity and resolu tion Sensitivity Resolution High Resolution Standard High Super Ultra High Binning Super Binning Ultra Binning Exposing a chemilumines cence and a digitized white epi illuminated image by sin gle click operation Add Digitization Image On Selecting detection method Method Chemiluminescence Fluorescence EtBr Fluorescence SYBR Green Fluorescence GFP Fluorescence Cy3 Fluorescence Cy5 Fluorescence Dy781 Fluorescence DAPI Digitization DIA Digitization EPI Changing tray position Method Tray position Select the tray position from 1 to 4 User Manual 28 9607 42 AB 123 A Appendix A 2 Quick reference functions 124 Function Function name Set item Focusing Fine adjust the light using Adjusting focus Brightness and the focus using Adjust Starting exposure Start Edit Preference Yes Inverting image data Digitization Epi illumination tab Invert pixel data Saving a file in TIFF format Edit Preference File Format tab Select a 16 bit Linear TIFF for mat
84. s size 250mm X 250mm 210mm X 140mm 180mm X 120mm 144mm X 96mm 105mm X 70mm Position Position Position Position Maximum capture size when a wide view lens is used TIP On the Epi tray there are round dents marked on the tray for positioning the sample Line up the sample using the appropriate dents For NP tray e The NP tray is to be used at tray position 3 Note The readable area will be slightly smaller using the F1 8 wide view lens 36 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 4 Placing the sample 4 Open the IDX door and insert the tray Place the Epi or Trans trays in position with the side with a hole facing outwards 5 Ensure the lens cap is removed and close the IDX door 4 5 Exposing chemiluminescent samples This section describes how to take images of chemiluminescent samples No illumination is used for this application Place a sample on the Epi tray Put the tray in the ImageQuant LAS 4000 then close the door F ImageQuant LAS 4000 File Edit View Option Help ImageQuant LAS 4000 Exposure Type 3 Exposure Time 4 O Auto Manual isec bd 5 Sensitivity Resolution Add Digitization image Oon Ed Method Tray position m Focusing Boon ImageQuant LAS 4000 User Manual 28 9607 42 AB 37 4 Operating the ImageQuant LAS 4000 4 5 Exposing chemiluminescent samples Step Action Click the Method T
85. scence Through Fluorescence EtBr Y UV Trans illumination _ W 605DF40 Digitization Epiillumination White Epi illumination Trans illumination White Trans illumination 105 x 70 mm a pm Prev Next Cancel Select the Method Light source Filter Iris and Tray position If FlatFrame Mode is set to manual in Edit Preference click the Next button Select the appropriate Flat Frame file Click the OK button FW N e ImageQuant LAS 4000 User Manual 28 9607 42 AB 75 4 Operating the ImageQuant LAS 4000 4 15 Exposing large sample using wide view lens Step Action Click the Focusing button Using for example some printed text on the sample tray turn the lens while viewing the screen and adjust the focus manually 3 ImageQuant LAS 4000 File Edit View Option Help ImageQuant LAS 4000 Exposure Type Sensitivity Resolution High Resolution Y Flat frame Auto Click the Return button TIP Click on the image to magnify it Click on the image again to return to the original size Select Exposure Type Exposure Type 5 v Precision Increment Repetition Program 6 Set the Exposure Time or Interval Time 76 ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 15 Exposing large sample using wide view lens Step Action Select Sensitivity Resolution Sensitivity Resolution L
86. sec 1 60sec 1 100sec Select Sensitivity Resolution Sensitivity Resolution L High Resolution Standard High Super Ultra High Binning Super Binning Ultra Binning TIP Select Help Sensitivity Resolution This opens the following diagram that de scribes the relation between sensitivity and resolution Sonsitivity Resotution ImageQuant LAS 4000 User Manual 28 9607 42 AB 45 4 Operating the ImageQuant LAS 4000 4 6 Exposing fluorescent samples EtBr Step Action Click the Start button The orange Busy LED lights on the instrument during exposure F ImageQuant LAS 4000 File Edt View Option Help ImageQuant LAS 4000 Exposure Type Exposure Time Auto Manual Sensitivity Resolution E Time High Resolution Y AT Dee Finishing Time AM 09 40 6 Exposing TTT Flat frame Auto Cancel Fie E amp R Vew Option Help ImageQuant Exposure Type Exposure Time Mto Manual pene Y nsitivity Resolution per Flat frame mso EJ Complete Click the Complete button The display returns to the main screen TIP If the Method EtBr UV Trans illumination is used the Start button can be clicked and exposure taken even if the temperature setting state of the CCD is Not Ready ImageQuant LAS 4000 User Manual 28 9607 42 AB 4 Operating the ImageQuant LAS 4000 4 7 Exposing fluorescence Epi illumin
87. sssesssses 4 1 Layout of the ImageQuant LAS 4000 Control Software oonneeeeessssesseseesesesseneees 4 2 Preparation TOF EXDOS TE sisgiicinnsane anina n unninn 43S CABANON aranana TE TEENE AA Placing te SAMPE 5 1s essnssssstoavacisenesseneanennsetierneanernaneeanieennennmmnneeemmNnINERNE 4 5 Exposing chemiluminescent samples ou sssseseeesessecccesesssstesseseeeecessssnnnessseeceeeeeeennetes 4 6 Exposing fluorescent samples EtBr si i si scissecsseccctadsaccescs asnnss nsasebinesersstateesctassessicboive 4 7 Exposing fluorescence Epi illumination ccssccscssscssesssatescorssrosnssavinetccnencosossscoqveneoconvovbs 4 8 Exposing dye stained samples and films White Epi light 0 eeeececsscesseeeeee 4 9 Exposing dye stained samples and films Trans illumination sees 4 10 Exposing consecutively Increment s aicasciscsaciesdtinstaiitaatiatasanancanananaie 4 11 Exposing repeatedly Repetition scscsinsssssseseaseccnsosssaseivaeeescdeasnsunnperrnacocossonvpsesesevesescoasswoes 4 12 Printing exposed WMOGES icssssseces ccssessannsanennenennsieenannnemasaimeinnUneN 4 13 Saving exposed MMAGES cccecsuiiozsscrnepisssevasivenntetesonsssiuauietenepdiaseainreneesscnenonaeeienianessisoenete ATA ENAN INE SESSION asainne Gaeta ee aE 4 15 Exposing large sample using wide view IONS Q eccsssssssssssssesesccessssssssesesesesecessessees 4 16 Creating anew method and performing flat frame calibration 5 Software reference QUI ices sitcarcsscncccssdetirc
88. strator password if prompted then click Continue to proceed with the installation Step Action 1 Insert the ImageQuant LAS 4000 Control Software CD 2 Locate and double click the file ImageQuant LAS 4000 msi 3 In the ImageQuant LAS 4000 InstallShield Wizard dialog click the Next button 4 Read the license text If the license agreement is not acceptable please contact a GE Healthcare representative See the back cover of this manual for contact information Select accept the terms in the license agreement and click the Next button ImageQuant LAS 4000 User Manual 28 9607 42 AB 6 Installing ImageQuant LAS 4000 Control Software 6 3 Install ImageQuant LAS 4000 Control Software under Windows Vista Step Action 5 Select destination folder in the dialog T imageart AS 1000 atalsa cad cel Destination I older Chek Newt to naal to this Falder er chick Change to instal to a different Folder Install ImageQuant LAS 4000 to C Program Fins Change Cea a er e Click the Next button to install the software at the default folder C Program Files e Click the Change button to install to a different folder 6 Click the Install button 7 If User Account Control UAC is enabled in Windows Vista a dialog displays the message An unidentified program wants access to your computer Click Allow 8 Click the Finish button The installation of ImageQuant LAS 4000 Control Software is now completed 9 Start the ImageQua
89. th the equipment 2 In EU countries The cable is given the following CEE Certification Number VDE 6522 1570 8035 AIG DEMKO 116773 ASC SEMKO 9449029 KEMA 93 6830 04 KCS LB NEMKO P94102620 CEBEC 8652 El FIMKO 179615 02 OVE 0204 515 00 ImageQuant LAS 4000 User Manual 28 9607 42 AB F 1 Introduction 1 1 Important user information Mercury containing products label CAUTION A N K Ultraviolet ray lamps in this product contain mercury which must be lt recycled or disposed of in accordance with local state or federal laws LED safety This product is categorized as a class 1 laser LED device IEC60825 1 A2 2001 LED light sources Red Epi light Wavelength 630nm Green Epi light Wavelength 520nm Blue Epi light Wavelength 460nm RGB module Wavelength 630 520 and 460nm UV Epi light Wavelength 365nm NIR Epi light Wavelength 710nm Class 1 Class 1 Class 1 Class 1 Class 1M Class 1 White Epi light Wavelength 470 740nm Class 1 CAUTION Do not look at the light directly through optical instruments CAUTION If the door is opened and the interlock is cancelled class 1 laser LED will be emitted CAUTION gt gt e Use of controls or adjustments or performance of procedures other than those specified in the user documentation may result in hazardous radiation exposure Notes and tips Note A Note is used to indicate information that is important for trouble free and
90. thod Light Filter ECL Chemiluminescence none Through 0 85 ECL Plus Chemiluminescence none Through 0 85 Lumi Light Plus Chemiluminescence none Through 0 85 Renaissance Chemiluminescence none Through 0 85 Super Signal Chemiluminescence none Through 0 85 Bright Star Chemiluminescence none Through 0 85 CDP Star Chemiluminescence none Through 0 85 CSPD Chemiluminescence none Through 0 85 ImageQuant LAS 4000 User Manual 28 9607 42 AB 125 A Appendix A 3 Detection Reagents and corresponding settings ImageQuant LAS 4000 setting Classification Reagent name Method Light Filter Iris EtBr Fluorescence EtBr UV 312nm 605DF40 2 8 Trans Cy2 Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Epi SYBR Green Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Epi SYBR Green II Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Epi SYBR Gold Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Epi SYPRO Ruby Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Epi SYPRO Orange Fluorescence SYBRGreen Blue 460nm Y515 Di 0 85 Epi SYPRO tanger Fluorescence SYBRGreen Blue 460nm Y515 Di 0 85 ine Epi FITC Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Fluorescence dye Epi FAM Fluorescence SYBR Green Blue 460nm Y515 Di 0 85 Epi EGFP Fluorescence GFP Blue 460nm 510DF10 0 85 Epi ECFP Fluorescence GFP Blue 460nm 510DF10 0 85 Epi RITC Fluorescence Cy3 Green 520nm
91. tude 2000 m or lower Overvoltage category category Il Transient overvoltage Rated pollution applied Pollution Degree 2 Analysis unit interface USB 2 0 Do not connect the ImageQuant LAS 4000 USB connector to a computer not certified with UL60950 1 UL listed and IEC60950 1 Others Noise 70 dB A or lower ImageQuant LAS 4000 User Manual 28 9607 42 AB 131 A Appendix A 5 Minimum computer requirements A 5 Minimum computer requirements 132 Operating system Windows XP SP3 or Windows Vista Business SP1 32 bit Memory 1 GB or more Processor Intel Core 2 Duo processor HD 80 GB or more USB port version USB 2 0 Optical drives DVD ROM Monitor resolution 1280 x 1024 pixels or more ImageQuant LAS 4000 User Manual 28 9607 42 AB A 6 Service report fax sheet To SERVICE REPORT FAX SHEET Customer information 1 Company name 2 Person in charge 3 TEL 4 FAX 5 E mail ImageQuant LAS 4000 system information 6 Serial No e Camera head e Others 7 Analyzing unit type e Operating environment Windows e Machine model 8 Usage frequency 9 Trouble occurrence frequency 10 Error code and failure description ImageQuant LAS 4000 User Manual 28 9607 42 AB A Appendix A 6 Service report fax sheet 133 For local office contact information visit www gelifesciences co
92. v high Resolution Standard High Super Ultra High Binning Super Binning Ultra Binning TIP Select Sensitivity Resolution in the Help menu You can display Help that describes the relation between sensitivity and resolution iG 4 Click the Start button Exposure is started Adjust the gradations of the exposed image then save and print the image Click the Complete button 9 The display returns to the main screen ImageQuant LAS 4000 User Manual 28 9607 42 AB GE 4 Operating the ImageQuant LAS 4000 4 16 Creating a new method and performing flat frame calibration 4 16 Creating a new method and performing flat frame calibration For most purposes a pre existing method will be sufficient For other filter light source and iris combinations a new method needs to be created The following describes how to create a new method and perform a Flat Frame calibration Step Operation In Preference in the Edit menu select FlatFrame mode and set this to Manual 1 To create a new method click Method Setting in the Option menu The Method Setting dialog opens Method Setting Method List Name Light Filter Cy2 RGB E e Epi RG vsispi Fo 85 cy2 E Blue Epi Blue siso 2 Cy3 RGB E creen Epi RGB 5750F20 Cy3 E creen Epi Green 5750F20 Cy5 RGB E Red epi Re R670 Cy5 E Red Epi Red

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