Lumina Vision
Contents
1. Q Save Image Save your image window created in above onto your disk as an image file O From the File menu select the Save As command In the Save As dialogue box thus displayed specify the name of the file in which you want to save the image window by in this case enter ing TEST in the File name text box Then press the C button Chapter 6 Time lapse Image Capturing Optional Procedure 1 Standard Image Capturing Flowchart 2 Set Initial Image Capturing Condition 3 Set Initial Camera Condition 4 Set Camera Focus 5 Preview Image 6 Save Image Capturing Condition 7 Set Image Region to be Captured 8 Set Color Balance 9 Set View Range 10 Time lapse Capture Image 11 Save Image Chapter 6 Time lapse Image Capturing Procedure Chapter 6 Time lapse Image Capturing Procedure 9 Standard Image Capturing Flowchart In this chapter let s operate the LuminaVision s program in a similar man ner to that done in Chapter 5 to display the Image Capturing Palette window on the screen but this time to learn how to use a fluorescence camera to capture more than one piece of image into the system by specify ing the time for capturing each of the images according to the following flowchart D Set initial image capturing condition D Set initial camera condition 3 Set camera focus Preview image 6 Save image capturing condition Set image re2. 9 u SEC n 1 1039 Pinning 2 x 2 Exposure time 132 1 4 SEC n 1 x 106 9 u SEC n 1 519 Pinning 4x 4 Exposure time 132 07 u SEC Exposure time 238 95 u SEC Exposure time 238 95 4 SEC n 2 x 118 27 u SEC n 3 260 Frame blanking With frame blanking the exposure and image loading are repeated The exposure time can be set in the following ranges depending on the pinning No pinning Exposure time n X 111 2mSEC n 1 90 Pinning 2 x 2 Exposure time n X 55 6mSEC n 1 180 Pinning 4x 4 Exposure time n X 31 5mSEC n 1 325 6 8 Chapter 6 Time lapse Image Capturing Procedure 9 Preview Image After the camera focus setting is completed making sure that the camera focus has been set properly select the Preview image function to finalize the camera condition such as the exposure time and pinning In addition this function allows you to preview a color image captured into the system This function can be also used to make the final adjustment of the camera focus la e O When the image data to be captured is RGB color activate the RGB color check box in the Image Capturing Palette If your camera connected to the system is a monochrome one this check box cannot be selected Set Pinning by pressing its corresponding pop up button in the Image Capturing Palette You can set the pinning by selection from the three choices No 2X 2 and 4x 4 in which order the sensitivity increases al
3. Chapter 5 Image Capturing Procedure 9 Standard Image Capturing Flowchart In this chapter let s operate the LuminaVision program actually by display ing the Image Capturing Palette window on the screen to learn how to use a fluorescence camera to capture a piece of image into the system according to the following flowchart D Set initial image capturing condition D Set initial camera condition 3 Set camera focus Preview image 6 Save image capturing condition Set image region to be captured D Set color balance Set view range Si Capture Image Save image 5 1 Chapter 5 Image Capturing Procedure 2 Set Initial Image Capturing Condition 5 2 When you use a fluorescence camera to capture an image into the system display the Set initial image capturing condition dialogue box on the screen as shown in the following figure In the dialogue box set the image capturing method by selecting a camera according to the procedure described below From the tool bar select to display the Set initial image cap turing condition dialogue box AA ean E A a a a WB CoolShar Sansas at ORDA x In the dialogue box select the Capture image tab and set Method by selecting Camera In the Cameras or capture cards available for use with the system group box select the camera or capture card connected to the computer 4 Select the Co but
4. Image Data for Multicolor Composition 4 Save Multicolor Composite Image Chapter 7 Image Analysis Procedure 1 Chapter 7 Image Analysis Procedure 1 9 Analysis Flowchart In this chapter let s operate LuminaVision program to combine more than one image captured from a fluorescence camera into the system using a different excitation light from the one used in Chapter 5 to create one piece of image data according to the following flowchart D Load image daia D Combine image data for multicolor composition 3 Save multicolor composite image 7 1 Chapter 7 Image Analysis Procedure 1 Load Image Data Load the image data captured into the system according to the procedure described in Chapter 5 using a different excitation light to the one used in Chapter 5 In this case use a certain sample image data saved in the particular folder in the hard disk as described below O In the File menu click on the Open command to display the Open dialogue box on the screen From the LuminaVision folder displayed in the dialogue box select Img folder From this folder select image files sample_r sample_g and sample_b to load them 9 Combine Image Data for Multicolor Composition 7 2 O Open the Process menu From the menu select the Combine image data for multicolor composition command Specify the resolution of the image to be obtained by processing the three sample images for multicolor compositio
5. Normal Fixed at 55 6mSEC the exposure time cannot be changed Shutter mode With the operation of the electronic shutter the exposure and image loading are repeated The exposure time can be set in the following ranges depending on the pinning No pinning Exposure tine 132 1u SECH n 1 x 106 9u SEQ n l 1039 Pinning 2x 2 Exposure tine 132 1u SECH n 1 x 106 9u SEG n 1 519 Pinning 4x 4 Exposure tine 132 07u SEC Exposure tine 238 95u SEC Exposure ti ne 238 95u SECH n 2 x 118 27u SEG n 3 260 Frame blanking With frame blanking the exposure and image loading are repeated The exposure time can be set in the following ranges depending on the pinning No pinning Exposure tine nx 111 2N EQ n 1 90 Pinning 2x 2 Exposure tine nx 55 6nSEC n 1 180 Pinning 4x 4 Exposure tine nx 31 5n EC n l 325 5 7 Chapter 5 Image Capturing Procedure 9 Preview Image After the camera focus setting is completed making sure that the camera focus has been set properly select the Preview image function to finalize the camera condition such as the exposure time and pinning In addition this function allows you to preview a color image captured into the system This function can be also used to make the final adjustment of the camera focus he E O When the image data to be captured is RGB color activate the RGB color check box in the Image Capturing Palette If your cam
6. balance tab to set the white balance of the image Select the Specify white area by clicking button and use the mouse to click your selected white area in the image window This white balance setting allows the system to set the camera focus preview the image and capture it so that the clicked area will become white If after the clicking you want to further adjust the white balance of the image you can make the adjustment by using the R G B component adjusting sliders in the dialogue box Each slider has its left end set at 1 increasing its value when shifted rightward The value thus set will be added to its corresponding color component of the image data Chapter 5 Image Capturing Procedure Set View Range 5 14 If the camera captures a 10bit 30bit or above image you can use the Set view range function This function allows you to specify the intensity range within which you want to have your image data displayed in the image win dow Any image data whose intensity is out of the specified range will be set black or white The view range setting can be also achieved automatically during the camera focus setting image previewing and capturing processes by selecting the Auto scaling function provided in the Image Capturing Palette as shown in the following figure ele ol Ge A Fatz EZ ak ree E His zy Dr Al eee Nra O From the Image Capturing Palette select to display the Set view range dialogu
7. change graph by in this case entering TEST in the File name text box Then press hel ___ button X In this saving only the graph displayed in the window will be saved as an image data file an Select the List time series change data window In the File menu select Save to display the Save dialogue box as shown in the following figure In the Save dialogue box specify the name of the file in which you want to save the time series change graph by in this case entering TEST_DATA in the File name text box Then press the button X In this saving the data displayed in the window will be saved as an text data file 5 an erg D 8 7
8. for exiting LuminaVision You can cause the LuminaVision application to exit from the computer sys tem by selecting command Exit in the File menu Specifically the Exit command can be selected move the mouse cursor to File on the Menu bar at the top of the screen Click on the File menu to pull it down On the pull down menu thus displayed move the mouse cursor to command Exit and click on it 4 1 Chapter 4 Basic Operation Selecting Menu Items and Commands 4 2 The LuminaVision software program is designed so that its menu items and commands can be selected using the mouse in various ways such as Clicking quickly pressing and releasing its left button and dragging pressing and holding down its left button while moving it 1 Menu bar 774 SE Sr M Atr F t BH vroia ANT Clicking on one of these items on the Menu bar at its indicated position causes its corresponding menu to be displayed on the screen 2 Menu items 3 Tool bar commands Clicking on one of these tools on the Tool bar causes its correspond ing tool command to be executed 4 Dialo HER COBRA Zilles ziar Sip Chapter 4 Basic Operation gue box Control button EAR Bare AEE IHR H DEST Sagan EIFEL EI Command button Pop up button Command buttons Clicking on any of the command buttons causes its corresponding command to be executed Option buttons Option buttons are used fo
9. 64MB or above and a HD capacity of 6GB or above X The software requires the use of a Macintosh computer with a memory capacity of 128MB or above if installed with an optional function Time lapse image capturing 3 Keyboard 4 Display 5 Printer 2 2 Operating System OS The software requires the use of Mac OS8 5 or its upgraded version for its operating system 1 3 Chapter 2 Check before Power ON 1 Checking Product Contents 2 Connecting Copy Protection Device 3 Connecting Image Capturing Board Camera Chapter 2 Check before Power ON Chapter 2 Check before Power ON 1 Checking Product Contents This product contains the following items When you purchase the prod uct please check it to make sure that it contains all these items O LuminaVision CD R 1 floppy disk User s Manual 1 copy Instructions on Image Capturing Equipment 1 copy User Registration Card 1 copy Copy Protection Device 1 unit Connecting Copy Protection Device This software is designed so that even if installed in your computer it cannot be executed unless the copy protection device provided as its acces sory equipment is connected to the computer s USB port as illustrated in the following figure Macintosh USB HASP 2 1 Chapter 2 Check before Power ON 9 Connecting Image Capturing Board Camera 2 2 The image capturing board to be used should be kept in an anti stati
10. Lumina Vision USER S MANUAL Version 1 MITANI CORPOPATION PLEASE NOTE This manual whether in whole or part may not be reproduced in any form without the prior written permission of the corporation This manual is subject to change in its contents This manual has been prepared with full check to make sure of its contents being free of any error Notwithstanding this however if you find any error omission or doubtful point in the contents of the manual please contact our corporation Our corporation shall not be liable for any of the consequences of the use of the manual whether direct or indirect without reference to item 3 above Macintosh is the registered trademark of Apple Computer Inc This Lumina Vision software is based on CodeWarrior metrowerks Inc This product contains Stone Table TM by Stone Tablet Publishing All right to Stone Table TM and its documentation are retained by Stone Tablet Publishing PO Box 12665 Portland OR 97212 0665 USA Copyright C 1992 1999 Stone Tablet Publishing MITANI CORPORATION LV0003 Table of Contents Chapter 1 General Deserption cece eee eee 1 1 EE ee 1 1 2 Operating knveronment eee eee eee 1 2 2 1 Necessary Equipment GE 1 2 2 2 Operating System OS anca ela e aud 38 til 1 3 Chapter 2 Check before Power ON 2 1 1 Checking Product Contents ccccccceceeeeeeee sees 2 1 2 Connecting Copy Protection Device 0685
11. Measure the intensity of the ROI set in the stack window in O From the Analyze menu select Create time series change graph The Create time series change graph window will appear to display the measurement data on the intensity of the ROI in the stack window the type of which can be selected by clicking on the Type of data button from Minimum Maximum Total Average and Standard deviation The Create time series change graph window can normally accommo date up to 10 ROIs for display of their intensity measurement data If you want to display the measurement data for more than 10 ROIs in the window you can use the el button 8 5 Chapter 8 Image Analysis Procedure 2 9 List Time series Change Data Process the data displayed n for numerical representation in a tabular form O In the Create time series change graph window select the List data button When the List data button is selected the List time series change data window will appear as shown in the above figure 8 6 Chapter 8 Image Analysis Procedure 2 9 Save Time series Change Graph and Data Save the time series change graph and data obtained in E and 2 respectively O Select the Create time series change graph window In the File menu select Save to display the Save dialogue box as shown in the following figure In the Save dialogue box specify the name of the file in which you want to save the time series
12. Sc 3 Connecting Image Capturing Board Camera 2 2 Chapter 3 Software Installaton cece eee 3 1 1 Setting up System Software ccce eee eee 3 1 1 1 How to Set up LuminaVision EE 3 1 1 2 How to Set up HASP Driver EE 3 1 1 3 How to Set up Image Capturing Device Driver s ssesesseeseseessssese 3 2 1 4 Rebooting the Computer after Driver Setup s nsesssesesessonessssesesesee 3 2 Chapter 4 Basic Operation se cea oa oaia otet aici 4 1 1 Starting and EMI Gy escrire n 000 spate Aaa te ate 4 1 2 Selecting Menu Items and Command 4 2 3 Inputting Alphanumeric characters c00008 4 5 Chapter 5 Image Capturing Drocedure 5 1 1 Standard Image Capturing Flowchart 5 1 2 Set Initial Image Capturing Condition 5 2 3 Set Initial Camera Conditon eee 5 3 A Set Camera E EE 5 6 5 Preview AMA CE dea o use la lata EA AE 5 8 6 Save Image Capturing Condition eee 5 10 V Get Image Region to be Captured 068 5 11 EE Color Balance EE 5 12 9 Set View Range iii costes dela ata tisa aS atone ewan 5 14 NEW NEE 5 15 UT Say e Image ng sicsns doses eersten 5 15 Chapter 6 Time lapse Image Capturing Procedure 5 1 1 Standard Image Capturing Flowchart 5 1 2 Set Initial Image Capturing Condition 5 2 3 Set Initial Camera Conditon eee 5 3 A Set Camera Focus so eee ete o ae alea d ae deac 5 6 5 Preview EE 5 9 6 Save Ima
13. SenSYS Series connected to the computer and the reverse is also true Make sure that your camera selection is correct in this respect Chapter 6 Time lapse Image Capturing Procedure 9 Set Initial Camera Condition In the Image Capturing Palette you can set the initial camera condition such as the black level maximum minimum exposure time auto exposure condition and use disuse of the shutter in focus setting mode Eat Sets H D Va L GES Oasis O From the Image Capturing Palette select to display the Set Initial Camera Condition as shown in the following figure n Meu k e ee wu E CDS le Ee See Leta ERE DI 7 a 2 8 EED pyr iA 2 Set the black level of the camera Any camera captures some image if its lens opening is minimized Therefore the system must be set so as to exclude any image data captured from the camera with its lens opening minimized from the image data captured from it under normal condition for analy sis Such setting of the system can be achieved by capturing image data from the camera with its lens opening minimized and setting it as black according to the procedure described below In the Set initial camera condition dialogue box activate the Calibration check box and select the Measure button Then minimize the camera s lens opening for light shutoff If All preparations for the camera s black level setting are com pleted follow the message displayed on the
14. c electric bag before being actually connected to the computer When taking the image capturing board out of the bag for its connection to the computer hold its card edge never touching any of its components When connecting the image capturing board to the computer follow the procedure described in the manual provided with it uz For the latest information about the image capturing boards available for use with this software refer to Instructions on Image Capturing Equipment provided as a separate volume Chapter 3 Software Installation 1 Setting up System Software Chapter 3 Software Installation Chapter 3 Software Installation 9 Setting up System Software 1 1 How to Set up LuminaVision Close other application if opened on the screen Set the LuminaVision CD R disk in the drive unit Open the CD R program menu From the menu copy the LuminaVision folder onto the desktop screen of your computer using the drag and drop technique 1 2 How to Set up HASP Driver This software is designed so that it can only start its operation on your com puter after making sure that the copy protection device has been properly connected to the computer s HASP This requires the copy protection key HASP driver to be installed in your computer The driver installation can be carried out as follows O Set the LuminaVision CD R disk in the drive unit Open the CD R program menu From the menu cop
15. can be set by selection from 1 to 10 000 The Interval condition group box with two options Start lt gt Start and End lt gt Start is used to select whether to define the Time interval as the time duration from the camera capturing Start time to the next camera capturing Start time or the time duration from the camera capturing End time to the next camera capturing Start time as described in the following figures lt Description of Interval condition group box gt When option Start lt gt Start is selected the Time interval will be defined as follows Camera capturing Camera capturing Next camera capturing Start time End time Start time Time 5sec When option End lt gt Start is selected the Time interval will be defined as follows Camera capturing Camera capturing Next camera capturing Start time End time Start time 5sec i Time Chapter 6 Time lapse Image Capturing Procedure Select the format in which you want to have your image data cap tured into the system From the Image Capturing Palette select Bio display a list of options From these options select Stack window in this case as a typical format in which to capture your time lapse image data into the system The Stack window format allows your time lapse captured images to be created as one stack window which can display these multiple image windows c
16. cted You can also use the box portion of the scroll bar E by dragging it to scroll the image display Slider CA 4 4 In a dialogue box requiring input of a value selected from more than one a slider appears to let you select the value by dragging the slider bar W left or right to set it at that value Chapter 4 Basic Operation 9 Inputting Alphanumeric characters The LuminaVision software is designed to allow any necessary alphanumeric character input to be made as described below 1 Inputting numeric characters Clicking this area causes the cursor to appear at the I position for numeric character input allowing any numeric character to be entered from the keyboard with the entered numeric character appearing at this position 2 Inputting alphabetical characters Clicking this area causes the cursor to appear at the I position for alphabetical character input allowing any alpha betical character to be entered from the keyboard with the entered alphabetical character appearing at this position 4 5 Chapter 5 Image Capturing Procedure 1 Standard Image Capturing Flowchart 2 Set Initial Image Capturing Condition 3 Set Initial Camera Condition 4 Set Camera Focus 5 Preview Image 6 Save Image Capturing Condition V Get Image Region to be Captured 8 Set Color Balance 9 Set View Range 10 Capture Image 11 Save Image Chapter 5 Image Capturing Procedure
17. displayed in the dialogue box select Img folder From this folder select image files sample_t1 sample_t2 and sample_t3 to load them In the window menu select Stack to display the Stack dialogue box as shown in the following figure Ir PAPE 37 4k erth Spas sKoateas vot E CI e i Poses HE HIZ AHRI A Omg EL Ii er tir OK ned lt ll Enter STACK TEST in the file name text box and activate the Stack all applicable windows check box with a check mark put in it If all the setting is completed select the __ button The three image files will be stacked to create a stack window with a file name of STACK TEST as illustrated in the following figure Bert Lb Pd tel ma Chapter 8 Image Analysis Procedure 2 Set ROI O From the Tool bar select the button Set a circular ROI in the stack window created in The setting of the ROI can be achieved by placing the mouse cursor at its starting point and dragging the mouse to its ending point lt Further information about ROI gt You can use 1 7 gt ste ef A lin the Tool bar for ROI setting When you select one of these buttons by double clicking the mouse button the Set ROI dialogue box will appear on the screen as shown in the following figure He i Ei seven ok When you click on either 7 or the dialogue box gives you three op tions from which you can choose one as the method of setting the s
18. e box as shown in the following figure When you want to select the Auto scaling function to cause the sys tem to set the view range automatically deactivate the No auto scaling check box with no check mark put in it When you want to set the view range manually activate the No auto scaling check box with a check mark put in it Then enter the mini mum and maximum values of the view range you need in the Minimum and Maximum text boxes respectively The view range you can set varies with the number of bits of the image to be captured in the system If you want the system to optimize the maximum and minimum values of the view range you set manually select the Optimize button Chapter 5 Image Capturing Procedure Capture Image ele ei rat Ses e gt His ane Destin RR LLB a E i Get O Select the format in which you want to have your image captured into the system From the Image Capturing Palette select to display a list of options From these options select Image window in this case as a typical format in which to capture your image into the system The Image window format allows your captured images to be created as one image window If you select File from the options list displaying the Save file dialogue box you can save your captured image data as an image file From the Image Capturing Palette select to capture your image into the system in the Image window format selected in O above
19. e previewing you can set a ROI on the image to be captured in order to allow the camera to capture only the ROI image region into the system thereby accelerating the image captur ing operation Dri AEE O Select the button in the Image Capturing Palette The selection of this button causes the screen to display the Set image region to be captured dialogue box as shown in the following figure Du Ll EEE ETHE gt EB PR u a n e In the dialogue box set the region ROI of the image you want to capture into the system by specifying its top left and bottom right coor dinates as exemplified in the above figure The ROI setting can be made not only by the above mentioned method which involves the use of the dialogue box for numerical setting but also by the method which involves the direct use of the image window on which to drag the cursor to create a desired rectan gular ROI on the image Ifthe ROI setting is completed press the C button 6 12 Chapter 6 Time lapse Image Capturing Procedure 9 Set Color Balance When the camera captures a color image it is necessary to set the image so that it can be captured into the system with its color balance optimized This image s color balance optimization can be achieved using the Set color balance function to select its black and white areas on the image window for its black and white component specification based on which the system is to make its color correcti
20. em you are required to focus your camera on the image in a short time because it may undergo quick discoloration The LuminaVision software is designed to set the camera focus on an image to be captured into the system always using its monochrome image data with a binning of 2x 2 Binning is a technique by which to allow a camera to capture an image by using more than one pixel of the camera for one pixel of the image to in crease its sensitivity With a binning of 2x 2 for example the camera can capture the image with the image s pixel size reduced to 1 4 of the camera s pixel resolution thereby shortening the time required for transmission of the image data from the camera to the personal computer The binning is also effective in increasing the sensitivity of the image to light allowing its exposure time to be shortened with resultant control of its discoloration that may otherwise occur during its focus setting process O Set your fluorescent sample in a microscope LN S r From the tool bar select to display the Image Capturing Palette as shown in the following figure CSL DCE jae any Dn pepe ia POI CTL i Je IE In the Image Capturing Palette set No of bits to that of the camera connected to the computer Chapter 6 Time lapse Image Capturing Procedure Set Exposure time and select the el button When this button is selected the system will cause the camera to start capturing the image displaying the cap
21. era connected to the system is a monochrome one this check box cannot be selected Set Pinning by pressing its corresponding pop up button in the Image Capturing Palette You can set the pinning by selection from the three choices No 2x 2 and 4X 4 in which order the sensitivity increases allowing the exposure time to be shortened It is therefore recommended that if your image is subject to severe discoloration in a short time the binning should be set larger to allow it to be captured into the system in a time as short as possible It should be noted however that a larger pinning setting results in re duced resolution of the image the resolution of the image captured with a pinning of 2X 2 is reduced to 1 2 of that with no pinning The same is true when the image resolution achieved with a pinning of 2X 2 is compared to that with a pinning of 4x 4 O The pinning setting varies with the specification of the camera you use with the system Carefully read the instruction manual for the camera 5 8 Chapter 5 Image Capturing Procedure Set Exposure time and select in the Image Capturing Palette When this button is selected the system will cause the camera to start capturing the image displaying the captured image data in the image window continuously You can change the exposure time even while the system is operating for continuous display of the captured image data in the image window The exposure time can be sh
22. g Palette as shown in the following figure apr DO in pir i aa In the Image Capturing Palette set No of bits to that of the camera connected to the computer Chapter 5 Image Capturing Procedure Set Exposure time and select the button When this button is selected the system will cause the camera to start capturing the image displaying the captured image data in the image window continuously You can change the exposure time even while the system is operat ing for continuous display of the captured image data in the image window The exposure time can be shortened by shifting the slider bar leftward and lengthened by shifting it rightward If the image is dark lengthen the exposure time Conversely if the image is too bright shorten the exposure time X When you use CoolSNAP SenSYS series you can select whether or not to output the signal to actuate the shutter in the focus setting mode by activating or deactivating the Use shutter in focus setting mode check box in the Set initial camera condition dialogue box Adjust the Z stage to optimize the camera focus on the image When the camera focus setting is completed select again to stop the image capturing operation When the ORCA Series is selected the exposure time can be set in three modes as described in the following table The camera focus can only be set with a binning of 2x 2 ORCA exposure time setting modes Mode Description
23. ge Capturing Conditon eee 5 11 V Get Image Region to be Captured cceceeeee 5 12 8 Set Color Balance deasa dea oa caile dada la gege 5 12 Set View E EE 5 15 10 Time lapse Capture Image cccee eee 5 16 DEEN 5 18 Chapter 7 Image Analysis Procedure 1 8 6 1 LAn lysis e EE E 6 1 2 BE WER EE 6 2 3 Combine Image Data for Multicolor Composition 6 2 4 Save Multicolor Composite Image 6 5 Chapter 8 Image Analysis Procedure 2 0085 6 1 Terie yee BG WET EE 6 1 ere dmiaee Data octets E 6 2 BODO BOC i sarea see au sa a aa aia al il al sai NOE Da da 6 3 4 Create Time series Change Grapb 6 5 5 List Time series Change Data 6 6 6 Save Time series Change Graph and Data 6 7 Chapter 1 General Description 1 Software 2 Operating Environment Chapter 1 General Description Chapter 1 General Description Er Software The Lumina Vision software is a fluorescent image processing software designed integrating all relevant programs from capturing of fluorescent image to its analysis To execute all these programs the software is pro vided with commands in plenty but properly organized in a menu form so that they can be simply selected and executed using the mouse making it very easy to operate The software is thus designed to allow even begin ners to easily proceed with highly sophisticated image analysis on a step by step basis in an interactive manner In particu
24. gion to be captured D Set color balance Set view range Si Time lapse capture image Save image 6 1 Chapter 6 Time lapse Image Capturing Procedure 2 Set Initial Image Capturing Condition 6 2 When you use a fluorescence camera to capture an image into the system display the Set initial image capturing condition dialogue box on the screen as shown in the following figure In the dialogue box set the image capturing method by selecting a camera according to the procedure de scribed below O From the tool bar select Si to display the Set initial image capturing condition dialogue box Aaah lt gt oF HH Coolshar Sarara 2 ORLA In the dialogue box select the Capture image tab and set Method by selecting Camera In the Cameras or capture cards available for use with the system group box select the camera or capture card connected to the computer Select the C button after confirmation that all the setting is correct Move the cursor to the tool bar and click on Ca to display the Image Capturing Palette as shown on the next page The camera selection in the Set initial image capturing condition dialogue box does not involve its physical access to the camera connected to the computer but no more than its software setting Therefore in the Cameras or capture cards available for use with the system group box you can select ORCA Series even if you have CoolSNAP
25. ion for analysis Such setting of the system can be achieved by capturing image data from the camera with its lens opening minimized and setting it as black accord ing to the procedure described below In the Set initial camera condition dialogue box activate the Calibra tion check box and select the Measure button Then minimize the camera s lens opening for light shutoff If All preparations for the camera s black level setting are completed follow the message displayed on the screen to click on the button AAD Fb es BALET betes TRL TRE OES Lo F a 5 3 Chapter 5 Image Capturing Procedure 5 4 Return the camera s lens to its original opening and follow the mes sage displayed on the screen as shown in the following figure to click on the button ASS eb SET LL Le ee Fe O The execution of steps and mentioned above completes the setting of the camera s black level Be sure to perform this process if you use a camera to capture image data into the system irrespective of whether the data is a color or monochrome image 4 Set the maximum minimum exposure time of the camera This setting can be made using the Exposure time group box in the Set initial camera condition dialogue box to enter your desired X This setting is effective only for the CooISNAP SenSYS Series In the case of the ORCA Series the maximum minimum exposure time is to be set in three modes each of which has a certain all
26. ize of your rectangular or circular ROI Set standard size which allows you to use the mouse to set the ROI size optionally Set size with aspect ratio fixed and Fix size which allows you to set the ROI size in pixels When you select the dialogue box gives you four options from which you can choose one as the method of setting the length of your linear ROI Set standard length which allows you to use the mouse to set the ROI length optionally Fix length as horizontal line and Fix length as vertical line which both allow you to set the ROI length in pixels and Fix length and angle 8 3 Chapter 8 Image Analysis Procedure 2 8 4 When you select one of ie F for ROI setting you can use 3 or option to set such ROI as illustrate in the following figures ROI O be 5 V 1 2 3 1 When you click on 3 the mouse cursor will have added to it at its lower right position to allow you to create more than one ROI which can be connected to each other 2 When you click on option the mouse cursor will have added to it at its lower right position to allow you to specify the area you want to delete from your ROI for example to create a doughnut shaped ROI 3 When you click on 3 option the mouse cursor will have X added to it at its lower right position to allow you to create the overlapping ROI area as a new ROI Chapter 8 Image Analysis Procedure 2 4 Create Time series Change Graph
27. l text box as a level for the exposure time determining condition This level can be regarded as the intensity of the image data to be captured from the camera The Upper limit text box in the Auto exposure condition group box is used to enter the upper limit on the exposure time in the auto exposure mode When in the Image Capturing Palette the Select auto expo sure for image capturing check box is activated to perform image capturing operation in the auto exposure mode the system will increase the exposure time from the Min value to the value set in the Upper limit text box to determine the exposure time at which the maximum intensity of the image data exceeds the value set in the Level text box as the exposure time in the auto expo sure mode Chapter 6 Time lapse Image Capturing Procedure For example when you capture a 12bit monochrome image which ranges in intensity from 0 to 1023 with 800 entered in the Level text box the system will determine the exposure time at which the maximum intensity of the image data first exceeds 800 as the exposure time in the auto exposure mode Set whether to use the shutter in the focus setting mode by activat ing or deactivating the Use shutter in focus setting mode check box X This setting is effective only for CoolISNAP SenSYS Series 6 5 Chapter 6 Time lapse Image Capturing Procedure 4 Set Camera Focus 6 6 When you capture a fluorescent image into the syst
28. lar the software can be characterized by the following features 1 A screen window of Image Capturing Palette designed to allow fluorescent image to be easily captured from a camera into the system for analysis 2 A software function available to process monochrome image 16bit and color image 48bit data 3 A software function available to process image data for multicolor composition and an optionally available software function to time lapse capture image as an option and 4 A system design made to allow use of Macintosh s excellent user interface 1 1 Chapter 1 General Description 2 Operating Environment 2 1 Necessary Equipment 1 Image capturing board camera The software requires the use of a camera designed to capture dark field image The software is designed to support Cool SNAP SenSYS Series and ORCA Series The Cool SNAP and SenSYS Series are provided with image capturing boards designed for exclusive use with them The ORCA Series requires separate arrangement for Snapper Data Cell Macintosh as its image capturing board V For the latest information about the image capturing boards available for use with this software refer to Instructions on Image Capturing Equipment provided as a separate volume Image capturing equipment LuminaVision Image output equipment 1 2 Chapter 1 General Description 2 Computer Macintosh designed with a memory capacity of
29. lect the one suitable for capturing the image of your current sample loading it onto the screen for its prompt preview Chapter 5 Image Capturing Procedure 9 Set Image Region to be Captured At the stage of the camera focus setting and image previewing you can set a ROI on the image to be captured in order to allow the camera to capture only the ROI image region into the system thereby accelerating the image capturing operation E gt III ls ean D Eat e t D GET i A GEE Dn rr ia FANM Ce oo Is i O Select the button in the Image Capturing Palette The selection of this button causes the screen to display the Set image region to be captured dialogue box as shown in the following figure 404 HL IESI vue Boa In the dialogue box set the region ROD of the image you want to cap ture into the system by specifying its top left and bottom right coordinates as exemplified in the above figure The ROI setting can be made not only by the above mentioned method which involves the use of the dialogue box for numerical setting but also by the method which involves the direct use of the image window on which to drag the cursor to create a desired rectangular ROI on the image If the ROI setting is completed press the Co button Chapter 5 Image Capturing Procedure 9 Set Color Balance 5 12 When the camera captures a color image it is necessary to set the image so that it can be capt
30. lected as your camera the system outputs the signal to actuate the shutter during the image captur ing Preview your image data to make sure that they are OK before clicking on again to stop the image capturing operation Chapter 6 Time lapse Image Capturing Procedure 9 Save Image Capturing Condition Save the data about the image capturing condition set using the Camera tab in the Image Capturing Palette No of bits Binning RGB color Select auto exposure for image capturing Exposure time as shown in the following diagram iS dei E Daa Va HRS SC vz Da DE Cl Meier ia Je El O Click on the Set pop up button to display a list ofitems From the list select Create a new camera condition to save the data on the initial image capturing condition set in the Image Capturing Palette The image capturing condition thus saved will be added to the items of the list that is displayed by pressing the Set pop up button O Next time when you capture the image of a sample of the same type as the one you captured previously you can press the Set pop up button to display a list of items including the previously registered image capturing condition s from which you can select the one suitable for capturing the image of your current sample loading it onto the screen for its prompt preview Chapter 6 Time lapse Image Capturing Procedure Q Set Image Region to be Captured At the stage of the camera focus setting and imag
31. llows the system to set the camera focus preview the image and capture it so that the clicked area will become white If after the clicking you want to further adjust the white balance of the image you can make the adjustment by using the R G B component adjusting sliders in the dialogue box Each slider has its left end set at 1 increasing its value when shifted rightward The value thus set will be added to its corresponding color compo 6 14 Chapter 6 Time lapse Image Capturing Procedure Set View Range If the camera captures a 10bit 30bit or above image you can use the Set view range function This function allows you to specify the intensity range within which you want to have your image data displayed in the image window Any image data whose intensity is out of the specified range will be set black or white The view range setting can be also achieved automatically during the camera focus setting image previewing and capturing processes by select ing the Auto scaling function provided in the Image Capturing Palette as shown in the following figure HEE We RE ele lad Ge fa fit Eat ea D Ve Haz e zzy DG Al eee O From the Image Capturing Palette select to display the Set view range dialogue box as shown in the following figure Sem Le ot Ea ar ee gi d When you want to select the Auto scaling function to cause the system to set the view range automatically deactivate the No au
32. lowing the exposure time to be shortened It is therefore recommended that if your image is subject to severe discoloration in a short time the binning should be set larger to allow it to be captured into the system in a time as short as possible It should be noted however that a larger pinning setting results in reduced resolution of the image the resolution of the image captured with a pinning of 2X 2 is reduced to 1 2 of that with no pinning The same is true when the image resolution achieved with a pinning of 2x 2 is compared to that with a pinning of 4x 4 The pinning setting varies with the specification of the camera you use with the system Carefully read the instruction manual for the camera 6 9 Chapter 6 Time lapse Image Capturing Procedure Set Exposure time and select in the Image Capturing Palette When this button is selected the system will cause the camera to start capturing the image displaying the captured image data in the image window continuously You can change the exposure time even while the system is operating for continuous display of the captured image data in the image window The exposure time can be shortened by shifting the slider bar leftward and lengthened by shifting it rightward If the image is dark lengthen the exposure time Conversely if the image is too bright shorten the exposure time When the Preview image function is executed with the CoolSNAP SenSYS Series se
33. n In the Combine image data for multicolor composition dialogue box as shown in the following figure select the Set tab In the Size group box click on the pop up button to select the resolution of the multicolor composite image you want as the numbers of its horizontal and vertical pixels If you want to use the No of pixels H and No of pixels V text boxes to set your selected image resolution click on the Size pop up button to select Optional In this case set the Size pop up button to select 640 H x 480 V FAS ep HAX 640 H x 480 oie t a bv ARE Chapter 7 Image Analysis Procedure 1 Then in the Process image for multicolor composition dia logue box select the Select tab From the image windows thus dis played in the dialogue box select the images to be combined for multicolor composition FLT AD aoe Click on the Red pop up button to select image file sample_r as a red component to be combined with the other two components to create a multicolor composite image Click on the Green pop up button to select image file sample_g as a green component to be combined with the other two components to create a multicolor composite image Click on the Blue pop up button to select image file sample bas a blue component to be combined with the other two components to create a multicolor composite image This case involves the use of onl
34. ollectively at one time If you select File from the options list displaying the Save file dialogue box you can save your time lapse captured image data as files by specifying the file name in the dialogue box For example if you enter TEST in the file name text box in the dialogue box your time lapse captured images will be saved as files with file names TEST00000 TESTO00001 and so on It should be noted in this connection that if after this time lapse image capturing process is completed another time lapse image capturing is performed with the same file name entered in the Save file dialogue box the newly time lapse captured image data will overwrite the previously stored image data AF Lumine vigion a El 74 7 a From the Image Capturing Palette select to capture your image into the system in the Stack window format selected in above Chapter 6 Time lapse Image Capturing Procedure Q Save Image Save your time lapse captured stack window created in 10 above onto your disk as a file O From the File menu select the Save command 2 In the Save dialogue box thus displayed specify the name of the file in which you want to save the stack window by in this case entering STACK TEST in the File name text box Then press the button Lc a 9 em fe D AF as TERT Tar Laos L a CS Chapter 7 Image Analysis Procedure 1 1 Analysis Flowchart 2 Capture Image 3 Combine
35. on upon its capturing The Set color balance function cannot be selected when your camera connected to the system is a monochrome one EH 7 1 a cerne eee Om eis 2 D Va His zii Dn pepe ORI Lei i he E Ge O From the Image Capturing Palette select NN to display Set color balance dialogue box as shown in the following figures ECKER ECKER MIA bets za In the Set color balance dialogue box select the Black balance tab to set the black balance of the image Select the Specify black area by clicking button and use the mouse to click your selected black area in the image window This black bal ance setting allows the system to set the camera focus preview the image and capture it so that the clicked area will become black If after the clicking you want to further adjust the black balance of the image you can make the adjustment by using the R G B component adjusting sliders in the dialogue box Each slider has its center set at 0 the right and left sides of which represent its plus and minus values respectively The value thus set will be added to its corre sponding color component of the image data Chapter 6 Time lapse Image Capturing Procedure In the Set color balance dialogue box select the White balance tab to set the white balance of the image Select the Specify white area by clicking button and use the mouse to click your selected white area in the image window This white balance setting a
36. ortened by shifting the slider bar leftward and lengthened by shifting it rightward If the image is dark lengthen the exposure time Conversely if the image is too bright shorten the exposure time O When the Preview image function is executed with the CoolSNAP SenSYS Series selected as your camera the system outputs the signal to actuate the shutter during the image capturing Preview your image data to make sure that they are OK before clicking on Fail again to stop the image capturing operation 5 9 Chapter 5 Image Capturing Procedure 9 Save Image Capturing Condition Save the data about the image capturing condition set using the Camera tab in the Image Capturing Palette No of bits Binning RGB color Select auto exposure for image capturing Exposure time as shown in the following dia gram Va HRS any Oasis RARAN iti O Click on the Set pop up button to display a list of items From the list select Create a new camera condition to save the data on the initial image capturing condition set in the Image Capturing Palette The image capturing condition thus saved will be added to the items of the list that is displayed by pressing the Set pop up button O Next time when you capture the image of a sample of the same type as the one you captured previously you can press the Set pop up button to display a list of items including the previously registered image capturing condition s from which you can se
37. osite Image Save the multicolor composite image window created in In the File menu select the Save As command In the Save As dialogue box thus displayed specify the name of the file in which you want to save the multicolor composite image window by in this case entering MULTI in the File name text box Then press the button 7 5 Chapter 8 Image Analysis Procedure 2 1 Analysis Flowchart 2 Capture Image 3 Set ROI 4 Analyze Time series Change 5 List Time series Change Data 6 Save Time series Change Data Chapter 8 Image Analysis Procedure 2 Chapter 8 Image Analysis Procedure 2 1 Analysis Flowchart In this chapter let s operate LuminaVision program to set a ROI in the stack image window loaded onto the screen and measure the ROI in the image for its intensity according to the following flowchart D Load image data D Set ROI 3 Create time series change graph ER List time series change data 6 Save time series change graph and data 8 1 Chapter 8 Image Analysis Procedure 2 Load Image Data 8 2 Load the image data time lapse captured into the system according to the procedure described in Chapter 6 In this case use a certain sample image data saved in the particular folder in the hard disk as described below O In the File menu click on the Open command to display the Open dialogue box on the screen From the LuminaVision folder
38. owable range for the exposure time setting Set the camera lens exposure time determining condition for the auto exposure mode in which the system can automatically determine the exposure time while capturing image data from the camera Refer to Section 5 Preview Image This setting can be made using the Auto exposure condition group box in the Set initial camera condition dialogue box to enter 0 to 4095 in the Level text box as a level for the exposure time determin ing condition This level can be regarded as the intensity of the image data to be captured from the camera The Upper limit text box in the Auto exposure condition group box is used to enter the upper limit on the exposure time in the auto exposure mode When in the Image Capturing Palette the Select auto exposure for image capturing check box is activated to perform image capturing operation in the auto exposure mode the system will increase the exposure time from the Min value to the value set in the Upper limit text box to determine the exposure time at which the maxi mum intensity of the image data exceeds the value set in the Level text box as the exposure time in the auto exposure mode Chapter 5 Image Capturing Procedure For example when you capture a 12bit monochrome image which ranges in intensity from 0 to 1023 with 800 entered in the Level text box the system will determine the exposure time at which the maximum intensity of the image data fi
39. r any multiple choice item such as one shown above providing more than one option from which to select one for its setting Clicking on the option button O for the option you want causes a black dot to appear inside the circle of the button turning it into which indicates that that option has been selected Pop up buttons When any of the pop up buttons is selected it will display more than one option from which to select one In this case for ex ample clicking on the Grayscale causes a list of more than one option to appear from which you can select one you need by mov ing the cursor to it and releasing the clicking 4 3 Chapter 4 Basic Operation Check box a A check box is used for any command item that is to be set activated selected or deactivated unselected by clicking on the box to put a check mark in it turning it into or leaving it unchecked respectively Clicking on the checked box causes the selection of its corre sponding command to be cleared with the check mark deleted from it turning it into Check box MRF ri Ib iaz aw 1x Dre EZITA ELI i Cl LS IIe Scroll bar Appearing in the image window the scroll bar can be used to scroll the display of the image in it Clicking or dragging the H W position of the scroll bar allows the image display to be scrolled in the direction sele
40. rst exceeds 800 as the expo sure time in the auto exposure mode Set whether to use the shutter in the focus setting mode by activat ing or deactivating the Use shutter in focus setting mode check box This setting is effective only for CooISNAP SenSYS Series 5 5 Chapter 5 Image Capturing Procedure 4 Set Camera Focus 5 6 When you capture a fluorescent image into the system you are required to focus your camera on the image in a short time because it may undergo quick discoloration The LuminaVision software is designed to set the camera focus on an image to be captured into the system always using its monochrome image data with a binning of 2x 2 Binning is a technique by which to allow a camera to capture an image by using more than one pixel of the camera for one pixel of the image to increase its sensitivity With a binning of 2x 2 for example the camera can capture the image with the image s pixel size reduced to 1 4 of the camera s pixel resolution thereby shortening the time required for transmission of the image data from the camera to the personal computer The binning is also effective in increasing the sensitivity of the image to light allowing its expo sure time to be shortened with resultant control of its discoloration that may otherwise occur during its focus setting process OD Set your fluorescent sample in a microscope From the tool bar select Gr to display the Image Capturin
41. screen to click on the Co button AAS UF es PERLE betes TILT ke ES Leer 6 3 Chapter 6 Time lapse Image Capturing Procedure 6 4 Return the camera s lens to its original opening and follow the message displayed on the screen as shown in the following figure to click on the button AAS PS Pes PRET LE LE Lo EL CET O The execution of steps and OG mentioned above completes the setting of the camera s black level Be sure to perform this process if you use a camera to capture image data into the system irrespective of whether the data is a color or monochrome image Set the maximum minimum exposure time of the camera This setting can be made using the Exposure time group box in the Set initial camera condition dialogue box to enter your desired values in the Min and Max text boxes This setting is effective only for the CooISNAP SenSYS Series In the case of the ORCA Series the maximum minimum exposure time is to be set in three modes each of which has a certain allow able range for the exposure time setting Set the camera lens exposure time determining condition for the auto exposure mode in which the system can automatically deter mine the exposure time while capturing image data from the camera Refer to Section 5 Preview Image This setting can be made using the Auto exposure condition group box in the Set initial camera condition dialogue box to enter 0 to 4095 in the Leve
42. to scaling check box with no check mark put in it When you want to set the view range manually activate the No auto scaling check box with a check mark put in it Then enter the minimum and maximum values of the view range you need in the Minimum and Maximum text boxes respectively The view range you can set varies with the number of bits of the image to be captured in the system If you want the system to optimize the maximum and minimum values of the view range you set manually select the Optimize button Chapter 6 Time lapse Image Capturing Procedure 10 Time lapse Capture Image Ed ma i When the Time lapse tab is selected SENA KR umile j be E eee Ey appears in the Image Capturing DHT ei Palette at this position O From the Image Capturing Palette select the Time lapse tab The Image Capturing Palette will then change to such one as shown in the above figure In the Image Capturing Palette activate the Execute check box with a check mark put in it Set parameter Time interval by selection from 1 to 32 767 to specify the time interval at which to capture your image data into the system Parameter Unit can be set by selection from second minute and day to specify the unit of time for the value entered in the Time interval text box Param eter No of cycles is used to set the number of times you want the camera to capture your image data at the specified time interval The No of cycles parameter
43. ton after confirmation that all the setting is correct Move the cursor to the tool bar and click on Ca to display the Image Capturing Palette as shown on the next page The camera selection in the Set initial image capturing condition dialogue box does not involve its physical access to the camera connected to the computer but no more than its software setting Therefore in the Cameras or capture cards available for use with the system group box you can select ORCA Series even if you have CoolSNAP SenSYS Series connected to the computer and the reverse is also true Make sure that your camera selection is correct in this respect Chapter 5 Image Capturing Procedure 9 Set Initial Camera Condition In the Image Capturing Palette you can set the initial camera condition such as the black level maximum minimum exposure time auto exposure condition and use disuse of the shutter in focus setting mode Eat e D Va L DE Dn efort O From the Image Capturing Palette select to display the Set Initial Camera Condition as shown in the following figure Dag bts Farce ve Si Ge Laien EARLE Dan n3p Frati eee MR Set the black level of the camera Any camera captures some image if its lens opening is minimized Therefore the system must be set so as to exclude any image data captured from the camera with its lens opening minimized from the image data captured from it under normal condit
44. tured image data in the image window con tinuously You can change the exposure time even while the system is operat ing for continuous display of the captured image data in the image window The exposure time can be shortened by shifting the slider bar leftward and lengthened by shifting it rightward If the image is dark lengthen the exposure time Conversely if the image is too bright shorten the exposure time O When you use CoolSNAP SenSYS series you can select whether or not to output the signal to actuate the shutter in the focus set ting mode by activating or deactivating the Use shutter in focus setting mode check box in the Set initial camera condition dialogue box Adjust the Z stage to optimize the camera focus on the image When the camera focus setting is completed select again to stop the image capturing operation O When the ORCA Series is selected the exposure time can be set in three modes as described in the following table The camera focus can only be set with a binning of 2x 2 6 7 Chapter 6 Time lapse Image Capturing Procedure ORCA exposure time setting modes Mode Description Normal Fixed at 55 6mSEC the exposure time cannot be changed Shutter mode With the operation of the electronic shutter the exposure and image loading are repeated The exposure time can be set in the following ranges depending on the pinning No pinning Exposure time 132 1 u SEC n 1 x 106
45. ured into the system with its color balance optimized This image s color balance optimization can be achieved using the Set color bal ance function to select its black and white areas on the image window for its black and white component specification based on which the system is to make its color correction upon its capturing The Set color balance function cannot be selected when your camera con nected to the system is a monochrome one Fast Sets LELEH apy Dn pepe aia O From the Image Capturing Palette select D to display Set color balance dialogue box as shown in the following figures ECKER EEECER In the Set color balance dialogue box select the Black balance tab to Select the Specify black area by clicking button and use the mouse to click your selected black area in the image window This black balance setting allows the system to set the camera focus preview the image and capture it so that the clicked area will become black If after the clicking you want to further adjust the black balance of the image you can make the adjustment by using the R G B component adjusting sliders in the dialogue box Each slider has its center set at 0 the right and left sides of which represent its plus and minus values re spectively The value thus set will be added to its corresponding color component of the image data Chapter 5 Image Capturing Procedure In the Set color balance dialogue box select the White
46. y MacHASPUsbDD into the system folder s function expansion folder 3 1 Chapter 3 Software Installation 1 3 How to Set up Image Capturing Device Driver Install your computer with the driver attached to the image capturing board The driver installation step can be omitted if the operation of the software will not involve any control of the image capturing equipment If the operation of the software involves any control of the image capturing equipment be sure to install the driver in the com puter If the computer is not installed with the driver you cannot capture your image from the device into the system 1 4 Rebooting the Computer after Driver Setup If the installation of the HASP driver and image capturing device driver is completed be sure to reboot the computer thus allowing it to become ready for the operation of the software 3 2 Chapter 4 Basic Operation 1 Starting and Exiting 2 Selecting Menu Items and Commands 3 Inputting Alphanumeric Characters Chapter 4 Basic Operation Chapter 4 Basic Operation 9 Starting and Exiting Procedure for starting LuminaVision O On the desktop screen open LuminaVision folder Double click on the LuminaVision application to start it When the loading of the application onto the computer is completed the computer display will show such a screen as shown in the following figure K H TI imi BE E ele E a AA Procedure
47. y three colors for multicolor compo sition However the software has been designed to allow the use of yellow magenta cyan and grayscale providing for seven channel multicolor composition O When steps are completed to select the three color compo nents for multicolor composition the resultant multicolor composite image will be previewed When you want to adjust the positions of the three images com bined their brightness and or contrast select the Adjust tab in the dialogue box VF AS SHK 7 3 Chapter 7 Image Analysis Procedure 1 7 4 Click on the Select component pop up button to select the color com ponent of the image you want to adjust for its position brightness and or contrast In this case set the pop up button to select the Red component To adjust the position of the color component use the iil E el gt buttons in the Move group box If you want to have the color component returned to its original position select the Hl button To adjust the brightness and contrast of the color component use their respective longitudinally arranged sliders in the dialogue box If all the necessary adjustment is completed and confirmed proper select The three color images will be combined as set to create a multicolor composite image window The multicolor composite image window is a 24bit color fixed image Chapter 7 Image Analysis Procedure 1 4 Save Multicolor Comp
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