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Time Lapse Imaging System BioStation IMQ CELL-S2 / CELL-S2

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2. Figure 1 1 3 Live observation screen Chapter 1 Basic Operations 1 1 2 Registering AF Position of the Reference Mark BS IM MC MOT Chamber When using the 5 MOT chamber with the AF mode enabled register the AF position of the reference mark as described below If the AF position of the reference mark is not registered the New time lapse setting screen cannot be displayed To carry out time lapse observation on the BS IM MC MOT chamber be sure to register the AF position of the reference mark 1 Check that the reference mark is displayed on the screen after the software is started and click the Mark registration button To make observations with greater AF accuracy check that the Stable is indicated on the screen display on the software When the Mark registration button is clicked the AF positions of the reference marks for wells 1 to 4 are registered in order This takes about four minutes When registration is complete an image of well 1 is displayed on the Live observation screen Figure 1 1 4 Reference mark register screen When you carry out only live observation click the Exit button the screen is switched to the Live Observation screen If the reference mark is not placed within the blue frame at the center of the screen click the Mark regist
3. 6 1 3 2 Checking the Device Information cod ooi ct Sce osi docens 7 15229 a aUe ed 7 tos Basics ellas aes Sates me 8 too HICSS OMUOM PAN lo duct Utama Ua uuo 9 Chapter2 Live Observation Screen oo o pe edo ta e ciue uu dva e rU du ou vu Vade ovv inne ied 10 2 1 Image Adjustment Buttons and Captured Image Display 11 2 1 Captured Image LISE View ce PONI a 13 2 01 2 14 2220 Checking S ALI ALION sx M 15 2 1 4 Intensity 16 Zoo sPSeudo XGolorDISDIBys os ees 17 2 2 Setting Observation Conditions Filter Magnification Mode Z position and Save 18 2 2 1 Saving Observation 20 2 3 Setting Observation Conditions Focus Mode Automatic Exposure Condition File Loading Light Intensity Exposure Time Gain and 21 2 3 1 Gain Compensation Value Fine 22
4. d 3000um cw amp 414 311 207 15 104x78um Simple Manual Load settings EPILamp C 100 4 Exposure Gain 100 4 640 x 480 Binning 15000 um Fix AE focus AE om 6 time Resolution Z Position 4 X 3000 33 v 300033 gt hm 150 00 um Live observation screen Figure 2 0 1 Table 2 0 1 Functions on the Live observation screen Item Live observation image display Observation point display Function The Live observation image of the field of view is displayed Click in this display to move the stage until the clicked point locates at the center of this display The field of view is indicated by the red frame and displayed as the live observation image Click the desired point to move the stage and to display the live image of the clicked point Image adjustment buttons Select to set tone adjustment saturation check intensity screening and pseudo color display Capture button Select to capture the live observation image and register it to the Captured image display 5 Captured image display Display area The images captured with the Capture button are thumbnailed Any thumbnail image can be enlarged and displayed on another window by clicking on the enlarge button magnifier icon Function To save the captured images highlight the thumbnail image and click the Image save butto
5. 535 575 500 H Color LJ the custam calor to averlap the image Custom color setting button Apply Figure 4 1 14 Filter settings dialog box Table 4 1 10 Functions for changing filter name display color and overlaid image color Item Function Filter The selected filter name is displayed Name Enter a filter name up to four alphanumeric characters EX DM BA Set the wavelength for the three filters Selecting the file name from the drop down list box of the Name box automatically apply the default wavelength and color to these three filters To change the wavelength adjust the value clicking the up or down arrow or V buttons or entering values Color Select a color from these buttons to apply its color to filter button displayed image color and graph line Click the desired color to display a check symbol Use the custom color to overlap the image Select to enable the custom color function with which you can set up the pseudo color for fluorescence images and the image color for overlaid image Custom color Select to display the Color dialog box You setting button can create an arbitrary color Basic colors The color can be used for the pseudo A color of fluorescence images and for the Tanta display color of overlaid images m mummmu Custom colors 255 40 Red 255 rTTTTTT AE Col
6. Ir E 88 Basic Operations This chapter describes the basic operations commonly used for starting and closing and all windows of the BioStation IM 1 4 Starting and Closing the Software This section describes how to start and close this software installed in the control PC 1 1 1 Starting the Software Starting the software from the shortcut icon on the desktop 1 Double click the Bio Station IM shortcut icon on the desktop of the control PC The BioStation IM starts up Bio Station Il Figure 1 1 1 Shortcut icon of Bio Station IM Starting the software from the start menu 1 Open the Start menu and then click Bio Station IM The Bio Station IM starts up Ma Q Recent Documents e My Pictures 25 My Music 8 My Computer Control Panel Set Program Access and Defaults PAYAK 2 Printers and Faxes nternet v My Documents nternet Explorer cJ Windows Update Help and Support windows Media Player 2 Search 277 Run Log Off 9 Turn Off Computer Figure 1 1 2 Start menu All Programs For the BS IM C chamber the Live observation screen appears first when this software starts For the BS IM MC MOT chamber with the AF Auto focusing mode enabled the Reference mark register screen appears first For registering the AF position of the reference mark on this screen see Section 1 1 2 Registering AF Position of the Reference Mark BS IM MC MOT Chamber
7. Save Intensity data Format Filename Time lapse log Filename testsach0S0512_ timelapse Tu Mark comment Filename Tem Figure 4 1 43 Save images Single point dialog box Table 4 1 22 Functions of image data save settings for a single observation point 1 2 Item Function Folder Select a folder to save images er Click the E button to display the Browse For Folder dialog box and specify the occ destination to save 9 My Computer a 314 Floppy A Sw Local Disk C Bros 8 COMPAQ O Documents and Settings O Drivers 8 O hp O i386 mc Image New Folder v Make Mew Folder Figure 4 1 44 Browse For Folder dialog box Save image Format Select a format from six formats to save images TIFF Gray 16 bits Color 24 bits TIFF Gray 8 bits Color 24 bits BMP Gray 8 bits Color 24 bits PNG Gray 8 bits Color 24 bits JPEG Gray 8 bits Color 24 bits or AVI 69 Item Save image Chapter 4 Time lapse Images in Process Screen Table 4 1 22 Functions of image data save settings for a single observation point 2 2 Function settings Enabled when AVI is selected in the Format field Interval Time opecifies the frame rate number of frames displayed per second at which pictures taken are saved into an AVI file maximum frames 60 Total Time opecifies the time during which pictures taken are saved into an AVI file Th
8. E File name test Ix Sample name m m Cell name m TE Points TR v 1 1 Ph Sms 300ms FI2 300ms 20 1 2 Ph Sms FM 300ms 2 300ms 20 1 3 Ph Sms 300ms FI2 300ms 20 3 v 1 4 Ph Sms 300ms FI2 300ms 20 5 E Time Range Fit v Intensity Range Fit v 0 2 0 Ratio 0 0000 Calculate m 4 Nov 26 05 00 PM Nov 26 05 05 Number of rounds 4 Temperature log e Chamber 370 370 Water 380 380 Oh 38s Oh Dim 18s Oh 01m 54s Oh 02m 34s Oh 03m Das Oh 03m 47s Outside 260 260 deg Round 4 gt E A 0h 00m 005 PE Figure 5 2 1 Time lapse images Acquired screen Points display Table 5 2 1 Functions on the Time lapse images Acquired screen Points display Item Function 1 Screen switch Select to highlight filters for references The selected filter is checked and the button 2 Observation image display 3 Selecting observation points 4 Calculate button image is displayed Multiple filters can be selected When multiple filters are selected the selected filters overlapped image is displayed Images at different points can be displayed on the four observation image displays Additionally the ROI for intensity analysis can be set for each observation point Click in the observation image display to select an observation point The point number at the upper left of the observation i
9. ko 414521 Tum 207x156um 104x Figure 3 1 24 Selecting observation point 2 Open the Zstack tab and edit Z stack Ce Kar Register paints with Z ztack Step 0 50 um Down 2 m steps Up 2 From 1 00 um Ta 1 00 Z stack sequence z Meh Zeg 2 14 Mch z V1 4 pem Register Figure 3 1 25 Zstack tab 3 Click the Time lapse experiment point Points Time Cell name etc registration button to confirm updated Ete ense setting conditions 9 Step 050 Am Independent steps Down 3 steps Up 4 steps From 1 50 um To 200 um Z stack sequence z Meh Zy Mch z 14 Mch z V1 Load Save Figure 3 1 26 Confirming setting conditions 38 Chapter 3 New Time lapse Setting Screen 3 2 Wide Field Screen If a specimen is exposed to an excitation light during the observation point settings and the observation condition settings the deterioration speed of the specimen becomes faster To avoid it use a captured tiled image to set up observation points and conditions On the Wide field screen you cannot adjust the light intensity and the focusing condition Therefore adjust intensity and focus on the Live screen in advance BioStation IM Version 2 20 Build 140 qunm Time lapse images Temp Chamber 370 570 Water 380 80 Acquired Outside 260 unitdeg C Stable Y St
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12. 1 2 and 2 2 use pseudo color 2 use pseudo Time lapse log Figure 5 1 4 Export images and data Single point dialog box 85 Chapter 5 Time lapse Images Acquired Screen Saving into the Bio Station IM format Click the Save into Bio Station IM format ics ids button of the Save image dialog box to display the Save into Bio Station IM format ics ids dialog box Save into Bio Station IM format ics ids E3 Folder Filename test Save time range C AJ One shot of current time Neighbors of current time C Hound E to 25 Time 12 00 00 tn 2478 Save points Select ALL Points Point 1 Point 2 Paint 3 Point 4 Save tone curve processing parameter Save ADI information me e e Figure 5 1 5 Save into Bio Station IM format ics ids dialog box Table 5 1 3 Functions on the Save into Bio Station IM format dialog box 1 2 Item Function Folder Select a folder to save an image Click the button to display the Browse For Folder dialog box and specify the destination to save B 9 my Computer a 314 Floppy 5 Local Disk C D Bros 53 COMPAQ 22 Documents and Settings 73 Drivers hp i386 Image O New Folder Browse For Folder Make New Folder Figure 5 1 6 Browse For Folder dialog box Filename Input a file name for the time lapse re
13. 48 4 1 2 Scrolling an Enlarged 49 4 1 3 Setting a Clipping Range of the Displayed 49 2 14 JMagniticattobBUtlODi Pis hot Un 50 AAS 2Stack Display COMUOL 50 4 1 6 Time lapse Observation Images Display 2 0000 52 4 1 4 intensity GIdDI caue ance S acc tese aac less 56 4 1 8 Intensity Graph Displaying 57 4 1 7 Time Line Display and Image Playback 59 4 1 10 Time lapse Experiment Scheme Points 61 4 1 11 Time lapse Experiment Scheme Time 62 4 1 12 Time lapse Experiment Process 63 2 1 13 Captured Image Printing ora a 65 4514 Captured Image Saving e eh em oe Bc os ai a eed 66 41 15 Example Of a Saved Iisdem ierit es 81 82 Time lapse Images Acquired 4 4 83 Channels DIS Day ss wets sos nM 83 5 1 1 Saving Time lapse Result File as a New File and Saving a Captured Image 85
14. Click the Device information menu to display the Device information dialog box This dialog box allows you to check information about the microscope and camera connected to the control PC The screen illustrated below shows the case for using the BS IM MC MOT chamber Device information Status Microscope Version Error Status Figure 1 3 3 1 3 3 Main Controller Ergo Controller Fiber Illuminations Chamber Chamber Gover Filter Cover Main Controller Temperature Controller Fiber Illuminations Hone Main Controller Controller Gamera Head Main Controller Connected Mat connected Connected BS IM MG MOT DEMO vDEMUO v DEMO Connected D S Gr Mc 001 006 1 700 000 DEMO Device information dialog box Checking the Error Information Click the Error information menu to display the Error information dialog box The item whose checkbox is checked is the cause of the error If there is no error the Error information menu is disabled thus this dialog box cannot be displayed Difference of the error detection scope by the microscope in use When the BioStation IM is used as a microscope only the temperature sensor status can be detected Only the checkbox of Temperature controller communication error is displayed on the Error information dialog box Error information Error Status Ux 1 Initialize error Temperatur
15. Select to save the time lapse result file as a new file BioStation IM format button The Save into Bio Station IM format ics ids dialog box appears Export images and data Select to save the time lapse images captured at a single observation Single point button point as image file s The Export images and data Single point dialog box appears Export images and data Select to save the time lapse images captured at multiple observation Multi points button points as image files The Export images and data Multi points dialog box appears Export large images Select to save the time lapse images Wide field image as image Multi points button file s The Export large images Multi points dialog box appears Cancel button Select to close the dialog box Export images and data Single point dialog box and Export images and data Multi points dialog box Export images and data of Point 1 and 20x v Save time range Operations of the two Export images and data dialog boxes are the same as the Time lapse images in process screen A Neighbors of current time Refer to the following sections Chapter 4 Time lapse Images in Process Screen section 4 1 14 Captured Image Saving to P Image data save settings for a single observation point 1 2 2 2 Save channel Ph Image data save settings for multiple observation points
16. 2 4 Observation Point and Focus 23 Chapter3 New Time lapse Setting 5 44040 44 41 412 8 24 9 1 JLIVO SCROOTI atu ERE CE aU EE 24 3 1 1 Time lapse Confirmation 26 3 1 2 Observation Point Verification Display 27 3 1 3 Time lapse Experiment Scheme Points 28 3 1 4 Time lapse Experiment Scheme Time 30 3 1 5 Time lapse Experiment Scheme Cell Name etc 33 3 1 6 Time lapse Experiment Scheme Zstack 35 3 1 7 Setting and Editing Observation Conditions Setting for Observation Filter and Magnification Editing 2 37 232 Wade Field E c 39 3 2 1 Observation Point Verification Display 41 3 2 2 Browsing or Clearing the Tiled 42 3 2 3 Setting Observation Conditions and Moving the Stage 44 Chapter 4 4 1 4 2 Chapter 5 5 1 5 2 Contents Time lapse Images in Process 46 Channels DIS play sides EE 46 4 1 1 Image Magnification and
17. 8 Figure 4 1 11 Time lapse observation images display Table 4 1 8 Functions on time lapse observation image display 1 2 Function Each observation images are displayed respectively on each display area ll Select to change the filter name and color of each filter button Right click the filter button to open the sub menu Select the sub menu to display the Filter settings dialog box You can change the filter name and select an image color in this dialog box Also you can set up the pseudo color for fluorescence images and the image color for overlaid image Select to display a scale bar on the bottom left of observation image The scale length is 10 um and has three ticks at um 5 um and 10 um Select to automatically set the range The range of the intensity to be displayed is automatically set The highest intensity A of the displayed image is set as the upper limit C and the lowest intensity B is set as the lower limit D D B Figure 4 1 12 Intensity graph Select to set tone curve adjustment manually The Tone curve adjustment dialog box appears to set tone curve adjustment Select to cancel tone curve adjustment and return to the original screen 52 Chapter 4 Time lapse Images in Process Screen Table 4 1 8 Functions on time lapse observation image display 2 2 Item Function Saturation Select to display saturation point in red check button To clear
18. Grid Figure 4 1 23 Calculate button IL Time Range Fit Intensity Range Fit v FI121131 Fl2 641 Ratio 1 7644 Grid Oh 00 40s Oh 01m 20s Oh 02m 005 Oh 02m 39s Oh 03 19s Oh 03 59s Stream block Round 4 4 gt gt gt Figure 4 1 24 Displaying the intensity graph 4 1 9 stream black a Item A to B repeating mark Number of stream block Number of rounds Playback slider Step buttons Stream block forward buttons Pause button Slow playback buttons Playback button Fast forward button Marked image playback button Repeat partially button Chapter 4 Time lapse Images in Process Screen Round Time Line Display and Image Playback Operation ete T ih 0h O1m 595 Figure 4 1 25 Time line display and image playback control Table 4 1 13 Functions on time line display and image playback control 1 2 stream black 8 A Oh 11m 485 Function The repeating section set with the Repeat partially button is indicated The number of the stream block of the playback slider s current position is displayed This field is displayed when playingback of the image captured by stream The number of rounds of the playback slider s current position is displayed This slider shows the current play
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20. 6 7 8 9 10 Chapter 4 Time lapse Images in Process Screen Table 4 1 1 Functions on the Time lapse images in process screen Channels display 2 3 Item Function Clipping button Select to perform clipping on the image area The function is turned on or off each time the button is clicked Magnification Select to show the current magnification buttons Z stack display The Z stack display control is displayed when Register points with Z stack control is checked on the Zstack tab of the New time lapse setting screen The pull down menu shows the Z coordinate of the image taken and time lapse observation image display can be switched by making a choice The slider allows you to switch the image display along the Z coordinate Image oftime lapse Phase contrast microscopy image of time lapse experiment is displayed experiment Ph If phase contrast filter is not selected for observation condition its image is not displayed Image of time lapse Fluorescent microscopy image of time lapse experiment is displayed experiment F11 If the fluorescent filter F11 is not selected for observation condition its image is not displayed Image of time lapse Fluorescent microscopy image of time lapse experiment is displayed experiment F12 If the fluorescent filter FI2 is not selected for observation condition its image is not displayed Image of time lapse experiment Multi When two or more filters are select
21. Chamber 370 370 Water 380 980 H setting in procesa Acquired Outside 260 unit deg C Stable j Status display and settings of the microscope X 3000um 414 311 207x156um 104x78um Simple J Manual AEffocus Load settings EPI Lamp 100 4 Exposure time 300ms zu Gain 100 4 P Resolution 640 x 480 v Z Position 150 00 um Fix ul 57 wel Focus lindo B 2 2 pS 4 4 15000 Y 3000 33 am b 4 Y J Figure 1 2 1 Live observation screen 1 2 1 Screen Switch Buttons Click these buttons to select each function of the BioStation IM series BioStation IM Version 2 20 Build 140 awe New Time lapse images Time lapse images Live observation time lapse setting in process Item Live observation New time lapse setting Time lapse image in process Time lapse images Acquired Figure 1 2 2 Screen switch buttons Table 1 2 1 Functions of screen switch buttons Function Select to use the BioStation IM series as a microscope Select to perform time lapse experiment Select to start time lapse experiment after setting observation point observation condition and so on The time lapse experiment in progress is displayed This screen automatically appears after time lapse experiment starts on the New time lapse setting scre
22. This operation is also available with the X stage knob or the Y stage knob of the ergonomic controller Jog dial Coarse focus Well switch button This button is displayed only for the BS IM MC MOT chamber Select to switch the well to observe in the glass bottom 4 well dish 45 Time lapse Images in Process Screen This screen shows process of time lapse experiment When time lapse experiment is started this screen automatically appears Therefore this screen cannot be displayed by operating any buttons There are the Channels display and the Points display for the Time lapse images in process screen Table 4 0 1 Channels display and Points display Item Function Channels Four display areas are provided for one observation point Three filter images filters overlapped image and Ratio image can be displayed at the same time Points One display area is provided for one observation point Switch the display area to display each filter image and overlapped image Therefore up to four observation points can be observed at the same time 41 Channels Display 1 gt p images T Temp Chamber 370 570 Water 380 80 f 4 Outside 260 unit deg C Stable 2 2 7 1 2776 5 AeA Iw m 1 1 1344 19 4399 85 v Ph 414x311um 202 15 104x78um Focus aj m File
23. Time lapse Images in Process Screen 4 1 8 Intensity Graph Displaying Method Intensity graph of an image can be displayed during a time lapse experiment or by selecting an area by specifying an area using the ROI mark in the display area of a time lapse result file The procedure to show the intensity graph for the Channels display is different from that for the Points display Channels display 1 Select the area of image to display the intensity graph Drag left click and drag a marquee around the area for the intensity graph display When performed in one display area the same areas in other three displays are selected at the same time Image area setting for an intensity graph When only the phase contrast Ph filter is selected for the microscopy no image area can be set for an intensity graph because the intensity analysis is not necessary Note on setting the ROI mark ROI mark 29 86 rn 414x311um 207x156um 104x78um Focus EXER E x FELT US e n iz mia Multi Ch Ph 2 Ratio 2 ki i BEBE m 0 25 _ Time Range Fit Intensity Range Fit v Intensity 4096 Oh 00 43s Oh 01m 26s Oh 02m 095 Oh 02m 51s Oh 03 34s Oh 04m 17s Figure 4 1 18 Selecting the area of image to display an intensity graph e Inthe scroll mode or the clipping mode no ROI mark
24. changing observation conditions Table 3 1 13 Functions for setting or changing observation conditions Item Function Time lapse Select to register the observation points and conditions to the experiment point Time lapse experiment scheme registration button If registered observation condition setting is changed the setting is overwritten New registration Select to register observation condition for a new observation point setting change select menu Select to change observation condition of the existing observation point Filter checkbox Select the filter used for the time lapse experiment Filter button Select the filter button to display the image with the selected filter The blue frame appears on the selected filter button This operation is also available with the Observation method switches of the ergonomic controller Select to perform phase contrast microscopy with the diascopic illumination of the red LED illuminator built in the microscope Fluorescent filter is not used Select to perform the fluorescent microscopy using the selected filter with the illumination of external mercury lamp This button does not appear unless the HG precentered fiber illuminator is connected and turned the power on Magnification checkbox Select the magnification for the time lapse experiment Magnification Select the magnification for the observation button This operation is also available with the Magnification
25. clipping area is set the save confirmation Bio Station IM dialog box for the clipped image appears Click the OK button in the dialog box Then seriei doping area the Save image dialog box appears To Cancel cancel the save operation and return to the Time lapse images Acquired screen click th 5 1 2 Save confirmation Cancel button dialog box If a clipping area is not set this dialog box does not appear but the Save image dialog box appears Select to measure the fluctuation amount of the stage from a captured image and correct the image by shifting it in the X and Y directions In principle the image size is reduced by the shift amount The area shifted is marked with a black frame Corrected image data is saved in a new file 84 Chapter5 Time lapse Images Acquired Screen 5 1 1 Saving Time lapse Result File as a New File and Saving a Captured Image Click the Save button D or press the Ctrl S key on the keyboard shows the Save image dialog box When a clipping area is set and the OK button is clicked on the save confirmation dialog box for a clipping image the Save image dialog box appears Save Image Export images and data ingle point Export images and data Multi points Export large images Multi points Cancel Figure 5 1 3 Save image dialog box Table 5 1 2 Functions for saving time lapse result file as a new file and saving a captured image Item Function Save as
26. desired observation points uncheck the Select ALL Points checkbox and check desired observation points Save time range Select to save all images of the selected observation point under the selected magnification in a file One shot of current time Select to save only the currently displayed image in a file Neighbors of current time Select to save the currently displayed image the five preceding images and the five following images total of 11 images into a file Concatenate all marks Select to save all concatenated images into a file The concatenated images consist of images of all marks including user marks and cell stimulate marks added in the Time Line five preceding images of each mark and five following images of each mark If neither user mark nor cell stimulate mark is set this item is disabled 73 Chapter 4 Time lapse Images in Process Screen Table 4 1 24 Functions of image data save settings for multiple observation points 2 2 Item Function Save time range Input time range Round Saves the image in the specified round range as a file According to the round count specified in these fields the time of the Time field is automatically set Time Saves the image within the specified shooting time as a file According to the time specified in these fields the count of the Round field is automatically set Select to save the Ph channel data into a file Save channel 11 Select to save th
27. image in a file Neighbors of current Select to save the currently displayed image the five time preceding images and the five following images total of 11 images into a file Concatenate all marks Select to save all concatenated images into a file The concatenated images consist of images of all marks including user marks and cell stimulate marks added in the Time Line five preceding images of each mark and five following images of each mark If neither user mark nor cell stimulate mark is set this item is disabled Input time range Round Saves the image in the specified round range as a file According to the round count specified in these fields the time of the Time field is automatically set Time Saves the image within the specified shooting time as a file According to the time specified in these fields the count of the Round field is automatically set 67 Chapter 4 Time lapse Images in Process Screen Table 4 1 21 Functions of image data save settings for a single observation point 2 2 Item Function Save channel Ph Select to save the Ph channel data into a file Select to save the 1 channel data into a file 11 save the image with the pseudo color check the use pseudo color checkbox Select to save the FI2 channel data into a file 2 To save the image with the pseudo color check the use pseudo color checkbox Select to save overlaid image composed of any of the Ph Fl1 or FI2 c
28. image selected button Select more than one thumbnail image on the Captured image display 5 and click this button to add the overlaid image to the Captured image display 5 The Image overlay button is enabled only when the XY coordinates of selected thumbnails match and different filters are used Trash button Select to delete only selected thumbnail image The frame of selected thumbnail image is displayed in blue Close button Select to close the Captured Image Gallery window Right click menu Right click the thumbnailed image to display the menu In the menu saving and deleting the clicked images or selected images are available 13 Chapter 2 Live Observation Screen 2 1 2 Tone Curve Adjustment Click the Tone curve adjustment setting button to display the Tone curve adjustment dialog box Tone curve adjustment Figure 2 1 4 Tone curve adjustment dialog box Table 2 1 3 Tone curve adjustment functions Item Function Enable tone Select to enable the tone curve adjustment curve checkbox Apply button Select to apply the set tone curve adjustment setting to the Live observation image display Close button Select to close the Tone curve adjustment dialog box without applying the tone curve adjustment setting 14 Chapter 2 Live Observation Screen 2 1 3 Checking Saturation Click the Saturation check button Lal to display saturation point of the live observation image in red Ad
29. is displayed Setting area Set water temperature inside the humidifier water tank to be suitable for time lapse experiment Outside Current ambient temperature is displayed If room temperature is below 18 C or above 28 C a warning symbol appears and POWER lamp of the LED indicator blinks rapidly Stable When both the temperature inside the culture chamber and the water temperature inside Unstable the humidifier water tank reach the set value and stabilize the status display changes from Unstable to Stable And time lapse experiment becomes possible Also the STABLE lamp of the LED indicator on the microscope is lit Displaying the logarithmic graph of temperature change Click the Temp button to display the Temperature dialog box Temperature logging start at Tuesday July 15 2009 14 35 11 The time lapse experiment is running under a stable condition Time lapse running Temp Fluctuations An unstable condition is detected during the time lapse experiment 1h24s 2h48s 3h1m12s 4h1m36s Close Figure 1 2 4 Temperature dialog box Table 1 2 3 Functions on the Temperature dialog box Item Function Range Select time range horizontal axis of temperature graph Temperature Temperature changes inside of the culture chamber and the humidifier water tank graph and that of ambient temperature are shown Close button Select to close the Temperature dialog box 5 Chapter 1 Basic Operations 1 3 Con
30. mode left fluorescent filter right phase contrast filter Table 2 3 1 Functions for setting observation conditions 1 2 Item Function AE focus Select only for the Manual mode button The exposure adjustment is performed with a focus priority mode It is useful for the focus adjustment because the exposure time for the dark specimen is 1 6 second maximum However noise of the image increases because this operation increases gain In this case exposure time and gain are adjusted automatically These conditions cannot be adjusted manually Click this button again to clear focus priority mode AE focus AE button Select only for the Manual mode Automatic exposure is performed once Exposure time and gain value used for the automatic exposure are displayed in each item box of observation condition Load settings gs Click here to load the already registered observation condition file Intensity Adjust intensity of each lamp with the slider and the right and left triangle buttons AE setting Display changes depending on selected filter This operation is also available with the Illumination intensity adjustment selector UP DOWN switch of the ergonomic controller For fluorescent filter set the intensity of episcopic illumination external mercury lamp Select to open or close the shutter for excitation light EPI Lamp They are enabled only when fluorescent filter is selected This operation is also available wit
31. name test 7 Patio V 1 1 Ph Sms 200ms FI2 300ms 20x ete Stop 3 o 4 oo lA 4 d lA Nov 25 0443 Nov 25 0444 PM m j y Time line MEBEEEEEEEEEEREN Number of rounds 1 2 Time Range Fit m Intensity Range Fit Y Fl1 1131 Fl2 641 Ratio 1 7644 Passage of time Oh Olm 45s Oh 01 005 Ou 1 5 Intensity 2059 1919 capturing Pointt 1 24 FDMQQ 1 4 1750 4 Temperature log w Saturation n Qin 1 6 1641 1501 bes Pause Stop Oh Q0 m 405 Oh 01m 20s Oh O2m 005 Oh 02 39s Oh O3m 19s Oh O3m 59s 4 De 7 A Oh 00m 00 c2 Gm 1 2 asp J 22 JA 22 lle t i LW diy les 1 8 Figure 4 1 1 Time lapse images in process screen Channels display Table 4 1 1 Functions on the Time lapse images in process screen Channels display 1 3 Item Function 1 Screen switch select to switch between the Channels screen and the Points screen buttons 2 Image magnification Magnification of the current image compared to the original image size and enlarge reduce resolution is displayed Click the magnifier buttons to enlarge or reduce buttons the image 3 Scroll mode button Scroll the image freely by clicking this button and dragging the mouse pointer on the enlarged image 46 4 5
32. of the user marks and cell stimulus marks The suffix mark is added to the end of the file name automatically Select to save the image into a file according to setting Save succeeded After saving is succeeded the dialog box appears Click the OK button to close the dialog box Figure 4 1 45 Save succeeded dialog box Select to return to the Export images and data Single point dialog box Select to close the Save images Single point dialog box 70 Chapter 4 Time lapse Images in Process Screen How to write information into images Use the Save images Advanced settings dialog box that can be displayed with the advanced settings button If TIFF Gray 16 bits Color 24 bits is selected in the Format field writing information is disabled To enter the information on the images to be saved select Gray 8 bits Color 24 bits in the Format field Save images Advanced settings Image processing Apply tone curve processing Write current ROI into images Add ratio color bar only ratio image Overlay text Observation settings left upper Scale amp User Mark left lower Gell name etc ri amp ht upper Round information right lower Use font setting ow Arial Rounded MT Bold Batang PaE M 16 v Color Set Default Figure 4 1 46 Save images Advanced settings dialog box Table 4 1 23 Settings for writing information into image
33. only the image within selected area 1 Select browsing area Drag left click and drag a marquee around the area to browse on the tiled image display 2 Click the Browse button Click the Browse button to start browsing image within the selected area Display of the Browse button changes to Browse cancel while the image is browsed To stop browsing the image click the Browse cancel button 3 To select multiple areas repeat selecting the areas to browse and click the Browse button Figure 3 2 6 Selecting browsing area Browsing the entire image of observation area 1 Click the Browse all button Clear browse Browse Browse a 1 Click the B Il button to start b ing th iste erben E n ick the Browse all button to start browsing the entire image of observation area Display of the Browse all button changes to Browse cancel while image is browsed To stop browsing image click the Browse cancel button Figure 3 2 7 Starting browsing image 43 3 2 3 Chapter 3 New Time lapse Setting Screen Setting Observation Conditions and Moving the Stage New point Baa 414 311 WideF ield dx 403 Full 2x 4x 20x 40x 80x Well A 2 v 5 4 3 4 i x 3000 33 Y 300
34. point Position of observation point its X and Y coordinates loaded file name of and file information time lapse result file sample name and cell name are displayed 83 3 4 5 6 Table 5 1 1 Item Time lapse experiment result Temperature log button Save button Fluctuation correction button Chapter5 Time lapse Images Acquired Screen Functions on the Time lapse images Acquired screen Channels display 2 2 Function Total time start time end time of time lapse experiment number of rounds logarithmic graph of temperature changes and maximum and minimum values of temperature change are displayed Select to display the logarithmic graph of temperature change the Temperature dialog box during time lapse experiment For functions of the Temperature dialog box see Displaying the logarithmic graph of temperature change on page 64 Clicking the Temperature log button on the Time lapse images in process screen displays the current temperature status during time lapse experiment however clicking the Temperature log button on the Time lapse images Acquired screen displays the temperature status during time lapse experiment result Select to save a time lapse result file as an alias file or to save an image captured with the time lapse experiment into an image file The Save button on the Time lapse images Acquired screen has a shortcut key setting Ctri S key on the keyboard If a
35. select to display the live observation image with the pseudo color of the filter Select to display the Intensity Screening dialog box for setting intensity screening This function can be used only for the fluorescent microscopy Saturation check Select to display saturation point in red button FL1 amp And then adjust the intensity and gain until there is no saturation point e button This function can be used only for the fluorescent microscopy Capture button Select to capture the live observation image and register it to the Captured image display If intensity screening and image adjustment are performed the performed image is registered With the single image capture switch of the ergonomic controller this operation can be performed Image overlay button Select to overlay more than one thumbnail image selected Select more than one thumbnail image on the Captured image display 5 and click this button to add the overlaid image to the Captured image display 5 The Image overlay button is enabled only when the XY coordinates of selected thumbnails match and different filters are used 11 Chapter 2 Live Observation Screen Table 2 1 1 Functions of the image adjustment buttons and captured image display 2 2 Item Trash button Image save button Captured image display Gallery button Reference mark button Function Select to delete only selected thumbnail image The frame of sele
36. tone curve adjustment parameter is saved The image to which the tone curve adjustment applied is not saved Select to save positions of the ROI set for each observation point BioStation AN Save succeeded Select to save images into a file according to settings When the file is saved successfully a dialog box appears Click the OK button to close the dialog box Figure 5 1 7 Save succeeded dialog box Select to return to the Save image dialog box Select to cancel saving and close the Save into Bio Station IM format ics ids dialog box 87 5 2 Chapter5 Time lapse Images Acquired Screen Points Display On this screen one observation point occupies one display area A filtered image or an overlapped filtered image of the observation point can be displayed on the area when specified Up to four observation points can be observed at the same time Functions other than the way to display images are the same as those for the Time lapse images in process Screen Channels BioStation IM Version 2 20 Build 140 test Temp Chamber 370 70 Water 380 80 Live observation mna New time lapse setting Outside 260 tunitdeg Stable Points ex JA Iw EJ 1 1 3000 33 3000 33 um 1 e Fle Ratio _FIL FI2 4l4x3llum 207x156um 104 8 ke qos miam n 1 4 3000 33 21 59 88 um
37. 0 33 4 dial P 4 Y Figure 3 2 8 Setting observation condition Table 3 2 4 Functions for setting observation conditions and moving the stage 1 2 Item Time lapse experiment point registration button Filter setting checkboxes Filter buttons Selecting magnification Register Wide field image points in a batch checkbox Function m Select to register the observation point data and the observation Regis conditions for the browsing to the Time lapse experiment scheme Select the filter used for the browsing the image as a tiled image Select the filter button to display the image with the selected filter When selecting multiple filters the images overlaid with the selected filters are displayed Switching display while browsing an image is not available Filter buttons not selected when browsing the tiled image are disabled to select The blue frame appears on the selected filter button This operation is also available with the Observation method switches of the ergonomic controller Select to perform phase contrast microscopy with the diascopic illumination of the red LED illuminator built in the microscope Fluorescent filter is not used Select to perform the fluorescent microscopy using the selected filter with the illumination of external mercury lamp This button does not appear if the HG precentered optical fiber light source is connected and turned the power on On the Wide field scr
38. 2 3 4 Figure 2 4 2 Position of the well number 23 3 New Time lapse Setting Screen You can set and save observation points and observation conditions and perform time lapse experiment on this screen Two screens Live screen and Wide field screen are provided to set observation points and observation conditions for time lapse observation On the Live screen the live image is used for the settings On the Wide field screen a captured tiled image is used for the settings When using the BS IM MC MOT chamber with the AF mode enabled the New time lapse setting screen cannot be displayed unless the AF position of the reference mark is registered To carry out time lapse observation on the BS IM MC MOT chamber be sure to register the AF position of the reference mark For details on registering AF position of the reference mark see Section 1 1 2 Registering AF Position of the Reference Mark BS IM MC MOT Chamber Do not have an access to the time lapse result file for the target specimen during time lapse experiment Otherwise experiment data obtained while the file is being accessed may not be saved on the Time lapse result file Ex If you copy the time lapse result file for backup from Explorer during the time lapse experiment the experiment data obtained while it is being copied will not be saved on the file 3 1 Live Screen oet observation points and observation conditions checking the live image on this scr
39. 2 ch2 Multi ch4 Advanced settings The Save images Advanced settings dialog box appears button This allows you to configure for example information to be written into images to be saved For detail see How to write information into images on page 71 Save button Select to save the image into a file Im according to setting After saving is succeeded the dialog box AN Save succeeded appears Click the OK button to close the dialog box Figure 4 1 50 Save succeeded dialog box Back button Select to return to the Export images and data Multi points dialog box Cancel button Select to close the Save images Multi point dialog box 76 Chapter 4 Time lapse Images in Process Screen Image data save settings for Wide field image 1 2 Click the Export large images Multi points button of the Save image dialog box to display the Export large images Multi points dialog box Export large images Multi points select export data Target Point 1 1 and 20x Order Typ Preview cave time range All One shot of current time Neighbors of current time Concatenate all marks Input time range Round ta E Time ta Save channel iw Ph Ww use pseudo color FI use pseudo color e e C Figure 4 1 49 Export large images Multi points dialog box Table 4 1 26 Functions of image data save settings for Wide field image 1 2 Item Function
40. 88500 NIKON CANADA INC tel 1 905 602 9676 NIKON FRANCE S A S tel 33 1 4516 45 16 NIKON GMBH tel 49 211 941 42 20 NIKON INSTRUMENTS S p A tel 39 055 300 96 01 NIKON AG tel 41 43 277 28 67 NIKON UK LTD tel 44 208 247 1717 NIKON GMBH AUSTRIA tel 43 1 972 61 11 00 NIKON BELUX tel 32 2 705 56 65 NIKON METROLOGY INC 12701 Grand River Avenue Brighton MI 48116 U S A tel 1 810 220 4360 sales_us nikonmetrology com NIKON METROLOGY EUROPE NV Geldemaaksebaan 329 3001 Leuven Belgium tel 32 16 74 01 00 sales_europe nikonmetrology com NIKON METROLOGY GMBH tel 49 6023 91733 0 sales_germany nikonmetrology com NIKON METROLOGY SARL tel 33 1 60 86 09 76 sales_france nikonmetrology com NIKON METROLOGY UK LTD tel 44 1332 811 349 sales_uk nikonmetrology com ADDRESS WAAVYALYAR Bio Science RRR T 100 8331 1 12 1 BRB FA 03 3216 2375 INO BRR NA AUTLIVAR 03 3216 2360 RUSHOIIAV ATTY T SR ADT ACER PCS 1 12 1 AREE AF 03 3216 9163 S342 FL SSS AT T 060 0051 RE 1 RR 2 8 2 SR Ez 011 281 2535 NAA HLL A ERE 465 0093 f Eh mtt 3 86 7rAFE JU 2F 85 052 709 6851 SM 2 9 ILA PESESR Bj A x T 532 0003 X Bx rh JI KBR 3 3 31 LHe E FA
41. Disclaimer Nikon shall not be liable for any damages and problems on user side or on a third party side which may result from the use of this software 2 Notes on the control PC e Factory Setting The software comes with a control PC Do not use any other PC to operate the product A change to the OS environment factory setting may lead to incorrect operation The control PC must not be modified by the user e Power saving mode Use of the control PC with power saving mode turned on causes the time lapse to stop halfway Be sure to set power saving mode to OFF factory setting in using the PC Anti virus software Set real time scanning of anti virus software to OFF Real time scanning enabled may cause this software to run unstably 3 File compatibility Files created by the software of Ver 1 to Ver 2 0 can be read into the software of Ver 2 1 or later whereas those created by the software of Ver 2 1 or later cannot be read into the software of Ver 1 Additionally if a file created by the software of Ver 1 to Ver 2 0 is read into the software of Ver 2 1 or later it is automatically upgraded to the file format of the version currently used Once upgraded it cannot be read into the software of the version by which it was originally created 4 About the example screens used in the manual e This manual describes various operations in BioStation IM IMQ Referred to as the BioStation IM series together hereafte
42. Each time this button is clicked the function changes as follows set A point gt set B point gt repeat from A to B clear repeating section 59 Item User mark setting button Cell stimulus mark setting button User mark Cell stimulus mark Chapter 4 Time lapse Images in Process Screen Table 4 1 13 Functions on time line display and image playback control 2 2 Function Select to set the user mark The user mark is displayed at the current playback point or observation point Select to set the cell stimulus mark The cell stimulus mark is displayed at the current playback point or observation point Right click this button to display the submenu for assigning keys Space Assignable keys are Space Return Shift Return and Tab Select from these four keys And then the selected key can be used instead of the cell stimulus mark setting button If the None key is selected no key is assigned This mark is displayed at the point set with the Edit user mark setting button Delete Right click the user mark to display the submenu Figure 4 1 27 submenu Figure 4 1 26 submenu Select Delete from the submenu to delete comments Select Edit to display the Mark Comment Input dialog box with which you can input confirm or change comments Click the Apply button in the Mark Comment Input dialog box to apply the edited Semmai comments Comment 1 Left click the
43. Functions on the Zstack tab Item Function Register points Select to enable the Z stack function On the Time lapse images in process screen with Z stack and Time lapse images Acquired screen the Z stack display control is displayed Step opecify the travel amount step width in the Z direction Enter a value with the keyboard Minimum step 0 05 um The value can be changed with the arrow buttons in 0 1 um steps Independent steps Select to enable the UP steps function Down Up Specify the travel count step count in the Z direction Enter a value with the keyboard The value can be changed with the arrow buttons Acceptable range 1 to 49 steps Up to 99 steps can be set for the Z stack current observation point 49 steps for Down Up From To Total travel amounts in the Z direction plus and minus directions appear These values are calculated automatically from the step width and the step count Z stack sequence Captures images with all selected filters by Z position of an identical observation point Select to capture images by filters at an identical observation point in a short time The filters are switched whenever an image is captured thus it takes longer time to make observations Captures observation images in the Z direction together by selected filters Select to make observations in a short time with less filter switchovers Load button Select to load the saved time lapse experiment condition file Sav
44. Images in Process Screen 4 1 12 Time lapse Experiment Process Display Start time of time lapse Nov 15 05 42 Nov 15 05 52 Estimated end time of experiment time lapse experiment Time line Number of rounds B bl Passage af time Oh Om Us 0h 10m Item Start time estimated end time Time line Number of rounds Passage of time Temperature log button Pause button Stop button Now captured Paint Fl2 20 Temperature log vwy Pause Stop Figure 4 1 38 Time lapse experiment process display Table 4 1 17 Functions on the time lapse experiment process display Function Start time and estimated end time of time lapse experiment are displayed month date time Process of time lapse experiment is displayed with the bar graph Number of rounds of running time lapse experiment is displayed current number of rounds total number of rounds Passage of time lapse experiment time is displayed current passage of time total observation time The logarithmic graph of temperature change since the software is started the Temperature dialog box is displayed This dialog box is the same as the one that is displayed by clicking the Temp button shown in upper right of the window Select to pause the running time lapse experiment If suspended the Start time lapse button on the New time lapse setting screen changes from Start time lapse to Restart time lapse for restart Select to stop the run
45. M508E 119 5 2 2 Nikon Time Lapse Imaging System BioStation IMQ CELL S2 CELL S2 P Instructions Application Software Introduction Thank you for purchasing a Nikon product This instruction manual is written for the users of the application software of the Time Lapse Imaging System BioStation IM IMa To ensure correct usage read this manual carefully before operating the product No part of this manual may be reproduced or transmitted in any form without prior written permission from Nikon The contents of this manual are subject to change without notice Although every effort has been made to ensure the accuracy of this manual errors or inconsistencies may remain If you note any points that are unclear or incorrect please contact your nearest Nikon representative If you intend to use any other equipment with this product read the manual for that equipment too If the equipment is used in a manner not specified by the manufacturer the protection provided by the equipment may be impaired Microsoft Windows and Internet Explorer are registered trademarks of Microsoft Corporation in the U S and other countries Other product and company names mentioned in this manual are trademarks or registered trademarks of their respective owners Notes on Handling the Software Read the following before use To ensure correct and safe operation read this manual before using the product 1
46. Select export data Select the first observation point of a Wide field image to save Order Select the registration order of observation points in a Wide field image to save Usually use Type1 when the Register Wide field image points in a batch checkbox is enabled Use Type2 to Type4 when the points are registered manually one by one 1 Left top gt right top gt left bottom gt right bottom Type2 Left top right top gt left bottom right bottom turn back on even lines Type3 Left bottom gt right bottom gt left top gt right top Type4 Left bottom gt right bottom gt left top gt right top turn back on even lines Tf Chapter 4 Time lapse Images in Process Screen Table 4 1 26 Functions of image data save settings for Wide field image 2 2 Item Function Select export data X Y combo Select matrix like selection area of a Wide field image box It allows you to select any number of images when you want to save a combined image of observation points that are not adjacent Usually when using the Register Wide field image points in a batch checkbox to register an area the number of images is automatically selected for registration at the time the Target is selected Displays the array of observation points Save time range All Select to save all images of the selected observation point under the selected magnification in a file One shot of Select to save only the currently dis
47. a ze produkt musi by wyrzucany oddzielnie Ponizsze uwagi maja zastosowanie tylko dla uzytkownik w z Europy Ten produkt jest przeznaczony do oddzielnej utylizacji i powinien by dostarczony do odpowiedniego punktu zbieraj cego odpady Nie nale y go wyrzuca z odpadami gospodarstwa domowego e Aby uzyska wi cej informacji nale y skontaktowa si z przedstawicielem przedsi biorstwa lub lokalnymi w adzami odpowiedzialnymi za zarz dzanie odpadami Hul Eur pai orsz gokban rv nes Elk l n tett hullad kgy jt s jelz se Ez a jelz s azt jelenti hogy ezt a term ket elk l n tve kell gy jteni Az al bbiak csak az eur pai orsz gokban l felhaszn l kra rv nyes Ezt a term ket a megfelel hullad kgy jt helyen elk l n tve kell gy jteni Ne dobja ki h ztart si hullad kk nt e Tov bbi inform ci k rt forduljon a forgalmaz hoz vagy a helyi hat s g hullad kgy jt s rt felel s r szleg hez E Symbol pro odd leny sb r odpadu platny v evropskych zem ch Tento symbol znamen e tento produkt se odkl dat odd len N sleduj c pokyny plat pro u ivatele z evropsk ch zem Tento produkt se m odkl dat na m st sb ru k tomuto elu ur en m Neodhazujte spolu s dom c m odpadem e V ce informac o zp sobu zach zen s nebezpe n m odpadem v m pod p slu n m stn instituce JESEN EEA COVUAMRILY TISA 8 88 II BUR ed
48. adjustment iiti selector switch UP DOWN of the ergonomic controller Details about setting observation condition moving observation point and focusing 9 414x311lum 207x156um 104x78um The following operations are the same as those of the Live observation LI im e focus screen setting observation condition selecting observation point with the state iene settings Jog dial and focusing EPI Lamp 100 4 41 k Exposure time 800ms 1 1 7 Gain P Reference Chapter 2 Live Observation Screen c m Z Position 150 00 um Fix e Section 2 2 Setting Observation Conditions Filter Magnification Mode Z position and Save Section 2 3 Setting Observation Conditions Focus Mode Automatic Exposure Condition File Loading Light Intensity Exposure Time Gain m and Resolution amp 150 00 um Section 2 4 Observation Point and Focus Adjustment 3 Chapter 3 New Time lapse Setting Screen Editing Z stack Follow the procedure below to edit the Z stack of an existing observation point Note that editing is not available after the start of time lapse experiment 1 Display the observation point of which you Lead aa Save want to edit Z stack Register point Select an observation point to edit on the New registration setting change select menu
49. age display range 22740 Figure 3 1 3 Observation point verification display Table 3 1 3 Functions of observation point verification display eum Function When two or more observation points are set at points very near each other or at a same point the symbol appears by the number This example indicates that observation points from 1 to 5 are set at the overlapped point The registered observation point is indicated by the blue frame The number over the blue frame is the same as the point number shown on the Point tab The field of view is indicated by the red frame and displayed as the live observation image X and Y coordinates of the center are displayed 2f Chapter 3 New Time lapse Setting Screen 3 1 3 Time lapse Experiment Scheme Points Tab Item Observation point Delete button Clear button Load button Save button Warning mark M 1 Fh1 180s FIT 1 45 FI2 1 42 20s M 2 Fh 1 1805 FIT 1 4 FI2 1 45 20 Save Figure 3 1 4 Points tab Table 3 1 4 Functions on the points tab Function Select the Points tab to display observation conditions filter name exposure time and magnification for each observation point The number next to the checkbox is the point number indicating the sequence of registration For the BS IM MC MOT chamber the well number is prefixed to the number When time lapse experiment is performed only point selected with a check mark is ob
50. age format from TIFF JPEG BMP or PNG The resolution of the image is fixed In addition a combined Wide field image will be reduced so that it can fit a size of one Wide field area or smaller based on a long side Ex For a 2x2 matrix observation area a combined image will be scaled down so that one side of the image is equal to the long side of the original size image Same length ON Combined Wide field image Selected area 2x2 matrix 40 Chapter 3 New Time lapse Setting Screen 3 2 1 Observation Point Verification Display The current pointer location the registered observation area locations and the current image display area can be identified on the upper screen Y 3511 3511 3511 3711 3711 3711 Figure 3 2 2 Observation point verification display magnifications left Full middle 2x right 4x Table 3 2 2 Functions of observation point verification display Item Function Registered The registered observation point is indicated by the blue frame observation point The number on the blue frame corresponds with the point number shown on the Point tab Current pointer The red frame yellow frame on the tiled image display indicates the current pointer and provides the X and Y coordinates of the center Click the desired point to move the current pointer to the position Image display area The area of the tiled image is indicated by the purple large frame The purple frame does not appear when
51. and file format Back button Select to return to the Save image dialog box Cancel button Select to close the Export large images Multi points dialog box 78 Chapter 4 Time lapse Images in Process Screen Image data save settings for Wide field image 2 2 Click the Next button of the Export large images Multi points dialog box to display the Save large images Multi points dialog box Save large images Multi points Folder Documents and Settings 0000 My 22 Save Image Format Galor 24bit e zi B mm Filename test Save z stack images According focus setting i50 H rj Advanced settings soe eet Ses Figure 4 1 50 Save large images Multi points dialog box Table 4 1 27 Functions of image data save settings for Wide field image 1 2 Item Function Folder Select a folder to save images button to display the Browse For Folder dialog box and specify the destination to save My Documents 4 My Computer 8 31 Floppy A Local Disk C Bos i C Documents and Settings C3 Drivers C3 i386 B Image C3 New Folder Browse For Folder RE RES Make New Folder Figure 4 1 51 Browse For Folder dialog box Save image Format Select a format from six formats to save images TIFF Gray 16bit Color 24bit TIFF Gray 8bit Color 24bit BMP Gray 8bit Color 24
52. appears 18 Save button Select to save images captured with the time lapse experiment to an image file When a clipping area is set the save confirmation dialog box for the clipped image appears Click the OK button in the dialog box Then the Save image dialog box appears cancel the save operation and return to the Time lapse images in process screen click A Save images of the clipping area the Cancel button When a clipping area is not set this dialog CUM box does not appear but the Save image dialog box appears Figure 4 1 2 Confirmation dialog box 4 1 1 Image Magnification and Enlarge Reduce Or 3 13 Figure 4 1 3 Image magnification Table 4 1 2 Functions of image magnification and enlarge reduce Item Function Magnification Magnification of the current image compared to the original image size resolution is displayed Enlarge reduce Select to enlarge or reduce the magnification of displayed images by 5 buttons Enlarge reduce Click the down arrow button at the right of the magnification to display menu 389 theimage magnification list box Click a magnification to enlarge or reduce the image magnification 48 Chapter 4 Time lapse Images in Process Screen 4 1 2 Scrolling an Enlarged Image Figure 4 1 4 Scroll mode button Table 4 1 3 Function for scrolling an enlarged image Item Function Scroll mode Scroll the image freely by clicking this button and dragging the
53. art time lapse 8 1 Ph Sms 20x v 2 2 Ph Sms 20x 3 Clear browse area Browse Browse all E e ES New point oniy 8 5 ly 414 311 WideF ield LI 33 3 did Full 2x 4x 20x 40x 80x 1 0 aS ara Seo SEs a gt EJ 439985 E ERSTE AG ERES 4 3 5 l Y 41 4x311 um 20x x Show cross line 5 1 1 Figure 3 2 1 New time lapse setting screen Wide field screen Table 3 2 1 Functions on the New time lapse setting screen Wide field screen 1 2 Item Function 1 select to switch between the Live screen and the Wide field screen buttons 2 Observation point The current pointer is indicated by the red frame The registered verification display observation point is indicated by the blue frame 3 3 Browselclear Select to set and clear a browse area of a tiled image buttons 4 Tiled image display The images browsed as a tiled image are displayed The current pointer is indicated by the yellow frame The registered observation p
54. back point and elapsed time and moves in synchronization with the playback image To play back the image from the point move this slider to a point with a mouse pointer or the right and left arrow buttons Select to play back the image forward or backward frame by frame Select to forward or backward to the start position of the stream block These buttons appear when the image captured by stream capturing is played back Select to pause playback of the image Select to slowly playback the image forward or backward One to three images per second The speed changes each time the button is clicked The number of dots blinking of the button indicates the speed Select to play back the image Five images per second Select to fast forward the image Six settings with skip intervals of 2 3 5 10 20 and 30 are available The reproduction speed changes every time the button is clicked The circle symbol of the button changes in accordance with the reproduction speed Select to play back ten or more images set before and after the user mark or the cell stimulus mark When the playback slider is on either mark images set before and after the mark are played back When the playback slider is not on either mark images set before and after the next mark are played back Select to set or clear a repeating section and to repeat the selected repeating section The A to B repeating mark appears on the selected repeating section
55. be registered 25 Chapter 3 New Time lapse Setting Screen 3 1 1 Time lapse Confirmation Window Click the Start time lapse button Start time lapse to display the following window Gonti mation window Total rounds 2 Total time Oh 01 s Point Kiumi Yum Filter Objective 1 1 2079 51 3079 51 Fh 2x 1 2 408026 215996 Fh ex 1 3 153283 3960 05 Fh ex start time lapse Prospective file size 351 MB Back to setting Figure 3 1 2 Confirmation window Table 3 1 2 Functions on the Time lapse confirmation window Item Function Start time lapse Select to display the Windows Save As dialog box button Input a file name for the time lapse result file and click the Save button And then the Time lapse image in process screen automatically appears and time lapse experiment starts Back to setting button Select to cancel time lapse experiment and return to the New time lapse setting screen 26 Chapter 3 New Time lapse Setting Screen 3 1 2 Observation Point Verification Display Checking the registered observation points and current view point are available in the entire observation area Click in the following screen to move the stage to the clicked point For the BS IM MC MOT chamber the Well switch tabs are displayed on the right side of the display Click the Well switch tab to see the observation points registered to each well Well switch tabs Y 22740 Item Registered observation point Live im
56. bit PNG Gray 8bit Color 24bit JPEG Gray 8bit Color 24bit AVI 79 Chapter 4 Time lapse Images in Process Screen Table 4 1 27 Functions of image data save settings for Wide field image 2 2 Item Function Save image settings Enabled when AVI is selected in the Format field Interval Time opecifies the frame rate number of frames displayed per second at which pictures taken are saved into an AVI file maximum frames 60 Total Time opecifies the time during which pictures taken are saved into an AVI file The number of frames and total time are calculated and if the frame rate is above 33 fps an error is indicated Frequency every Enabled when this checkbox is enabled Allowable range 2 to 99 The thinning out setting is valid for both Interval Time Total Time Filename Input a file name to save images Channel names are added to the end of file names automatically Ph ch Fl1 2 2 ch2 Multi ch4 Advanced The Save images Advanced settings dialog box appears settings button This allows you to configure for example information to be written into images to be saved For detail see How to write information into images on page 71 Save button Select to save the image into a file according ER to setting After saving is succeeded the dialog box A Save succeeded appears Click the OK button to close the dialog box Figure 4 1 52 Save succeeded dialog box Back butto
57. can be drawn on the image area Therefore cancel the scroll mode or the clipping mode to draw an ROI mark 2 Display the intensity graph Click the Calculate button The intensity graph of the selected area is displayed in the intensity graph display area When the intensity graph is displayed the selected area frame changes from yellow to red Display area of an intensity graph For information on the time horizontal axis and the intensity vertical axis of the intensity graph see Section 4 1 7 Intensity Graph 57 fu Calculate Figure 4 1 19 Calculate button e _ Time Range Fit Intensity Range Fit FI1 1131 Fl2 641 Ratio 1 7644 Grid wes 1 1 1 1 ian 1 Oh 00 40s Oh 01m 20s Oh 02m 00s Oh 02m 39s Oh 03m 195 Oh 03m 59s Stream block Round 4 4 gt gt gt 4 00m 00s Figure 4 1 20 Displaying the intensity graph Chapter 4 Time lapse Images in Process Screen Points view 1 Select the area of image to display intensity graph Drag left click and drag a marquee around the area for intensity graph display Different points and range for intensity graph display can be set to each observation point Image area setting for an intensity graph When only the phase contrast Ph filter is selected for the microscopy no image area can be set for an intensity graph because the intensity analysis is not necessary Note on
58. cted thumbnail image is displayed in blue Select to save all thumbnail images in a file Thumbnail image must be highlighted prior to saving Select save format from TIFF JPEG BMP or PNG For TIFF format image resolution is selectable between 16 bits and 8 bits The images captured with the Capture button are thumbnailed Click the Enlarge button magnifier icon at the bottom right of thumbnail image to display the enlarged image on other screen If number of thumbnail images becomes five or more a scroll bar appears at the bottom of thumbnail display Select to display the Captured Image Gallery window and to show the thumbnailed images captured with the Capture button in a list This button is displayed only for the BS IM MC MOT chamber Select to display the Reference mark register screen to register the AF position of the reference mark 12 Chapter 2 Live Observation Screen 2 1 1 Captured Image List View Figure 2 1 3 Captured Image Gallery window Table 2 1 2 Functions on the Captured Image Gallery window Item Function Enlarge button Select to display the enlarged image on other screen Image save Select to save all thumbnail images selected a file The thumbnail images button ig must be highlighted prior to saving Select the save format from TIFF JPEG or PNG For TIFF format image resolution is selectable between 16 bits and 8 bits Image overlay Select to overlay more than one thumbnail
59. cus graph dialog box Z focus graph Target Foint 1 1 and 20x 0 50um 1 m s zu m s 3 m s nm s Close Figure 4 1 10 Z focus graph dialog box Table 4 1 7 Functions on graphical display of focus positions Item Function Target Click the W button on the right side of the Target item to show the selection list of the observation points Selecting an observation point shows a graph of the focus positions registered for the observation point Close button Select to closes the Z focus graph dialog box 51 Chapter 4 Time lapse Images in Process Screen 4 1 6 tle Time lapse Observation Images Display Ph FI1 FI2 Multi Images of time lapse experiment are displayed If a filter Ph Fl is not selected for time lapse experiment scheme its image cannot be displayed Item Observation image display Filter button Scale bar button Automatic tone curve adjustment apply button Tone curve adjustment setting button Tone curve adjustment cancel button d re TRA T u T Je Fluorescent microscopy image 1 amp e 7 EN iJ po i fL E 2 it Fluorescent e microscopy image 2 LJ Lis Ls 8 Multi Ch Ph laj qu pv pr ie e 1 gt 1 1 Se d PD 41 x2 LT 3 12 ES 43 1 noXv T 3 i EI p Multi microscopy im LS
60. e 1 channel data into file To save the image with the pseudo color check the use pseudo color checkbox Select to save the FI2 channel data into a file To save the image with the pseudo color check the use pseudo color checkbox Select to save overlaid image into a file with the displayed Multi ch condition Next button Select to display the Save images Multi points dialog box In the dialog box specify the destination to save image and input its file name and file format Back button Select to return to the Save image dialog box Cancel button Select to close the Export images and data Multi point dialog box 74 Chapter 4 Time lapse Images in Process Screen Image data save settings for multiple observation points 2 2 Click the Next button of the Export images and data Multi points dialog box to display the Save images Multi points dialog box Save images Multi points Folder i ad e Documents Save Format 1 Gray Sbit Colorz24bit AVI settings f Interval Time i fps Total Time zec Iw Frequency every 2 frames Filename test_timells0e0 PhFIH FI SP md B Advanced settings e Ser Figure 4 1 48 Save images Multi points dialog box Table 4 1 25 Functions of image data save settings for multiple observation points 1 2 Item Function Folder Select a folder to save images Click the gal button to display the Browse For Fo
61. e Experiment Scheme Time Tab When the stream setting is configured during the time lapse experiment time setting the display of Time tab is replaced with Stream Acquisition cycle T atal time Rounds Oh 10m 00 20h 00m 0105 121 Delete Save Figure 3 1 10 Time tab Table 3 1 6 Functions on the Time tab Item Function Time lapse Select to display the capturing interval time total observation time and number of rounds experiment for time lapse experiment time display Click time lapse experiment time display to display the Timelapse dialog box for setting change In the dialog box changing and deleting capturing interval time and total observation time are available Delete button Select to delete the selected time lapse n experiment time The Delete confirmation dialog box appears A Delete from timelapse list To delete the selected time lapse experiment time click the OK button To return to the Time tab without deleting click the Cancel button Figure 3 1 11 Delete confirmation dialog box New button Select to set new time lapse experiment time The Timelapse dialog box for new registration appears Clear button Select to delete all settings of time lapse experiment time The Delete all confirmation dialog box AN appears To delete all settings of time lapse experiment time click the OK button To return to the Time tab without deleting clic
62. e button Select to save the registered observation point observation condition and time lapse experiment time as a time lapse experiment condition file Caution on the Z stack sequence e When switching the settings for 2 I j 4ch Z 1 after registering the observation points the Z stack sequence of all m is s changed e The image of an observation point registered with the Z stack function cannot be reproduced successively with this application software 35 Chapter 3 New Time lapse Setting Screen Z position display examples for observation points registered with the Z stack function The Point information dialog box for an observation point configured with Z stack shows the Z position as shown below Reference value negative side absolute value positive side absolute value number of steps Point information 414x311 um DIA Lamp Exposure time ain Resolution 640 x 480 Binning Dish Do I 074 51 um 3074 5 um test Delete Figure 3 1 22 Z position examples 36 Chapter 3 New Time lapse Setting Screen 3 1 7 Setting and Editing Observation Conditions Setting for Observation Filter and Magnification Editing Z stack NS NS Register Register AN v E v e For point 1 1 For paint 1 2 FI Phi For point 1 3 uU p 41421 207x156um 10478um 4l4x3llum 207x156um 104x um Figure 3 1 23 Setting or
63. e controller communication error Internal shutter error Initialize error Status 0x07 M matar error motor error motor error Objective motor error Filter cassette motor error intemal shutter error Running error Status 0 00 FAN motor error Heater error Temperature control error 2C bus error Mechanical black sensor error Figure 1 3 4 Error information dialog box BioStation IMa Chapter 1 Basic Operations 1 3 4 Basic Settings Click the Preferences menu to display the Preferences dialog box Preferences Preferences General General Data input when capture image saved Explorer launches when image capture Explorer launches when image capture Explorer launches when file export Explorer launches when file export ANE Viewing Viewing Exposure Time msec f Fractional Exposure Time msec f Fractional EPI Lamp v Transmission rate f Position EFI Lamp Transmission rate Position Bis IM MC chamber 4 Enable AF Mode Type Nomal H4 Launch as Mark window mode Cancel Cancel Figure 1 3 5 Preferences dialog box Figure 1 3 6 Preferences dialog box BS IM C chamber BS IM MC MOT chamber Table 1 3 2 Functions of Preferences dialog boxes 1 2 Item Function General Data input when Select to save captured images together with information such as capture image saved observation conditi
64. e number of frames and total time are calculated and if the frame rate is above 33 fps an error is indicated Frequency every Enabled when this checkbox is enabled Allowable range 2 to 99 The thinning out setting is valid for both Interval Time Total Time Filename Input a file name to save images Channel names will be added to the end of file names automatically Ph ch1 Flt ch2 2 ch2 Multi ch4 Save z stack Enables only the Z stack images Select to enable the images Z stack save settings According focus setting Saves images at the Z position registered by the Z focus mark button According range setting Saves images including Z stack data By changing the values of the From To fields the Z stack range to save the image can be specified Advanced Displays the Save images Advanced settings dialog settings button This allows you to configure for example information to be written into images to be saved Save Intensity data Select a format from the pull down menu to save the Format intensity graph Time lapse log Mark comment Save button Back button Cancel button Input a file name to save the intensity graph Filename The suffix intensity is added to the end of the file name automatically Input a file name to save a log file text file of time lapse experiment The suffix timelapse is added to the end of the file name automatically Input a file name to save comments
65. ed their images are overlapped and displayed Ch select to display the Ratio image of time lapse experiment Ls Bh Fa Select to display the Ratio setting dialog box and set FI FI2 EE the Ratio image 11 Intensity graph This graph shows changes in intensity during time lapse experiment The color of graph line is the same as that of each filter 12 13 14 Time line display Use the slider to check process of time lapse experiment The lt P button and image playback at the right and left of the slide bar is used to reproduce images step by control step or reproduce images step by step in a reverse order Also use the image control buttons to pause playback forward fast playback slowly reverse slowly repeat partially and mark a point Observation point The observation point of time lapse experiment is indicated by the yellow verification display frame The registered observation point is indicated by the blue frame Time lapse Points tab Select to display the observation conditions filter name experiment scheme exposure time and magnification for each observation point Click an observation point to display the Point information dialog box In the dialog box checking changing and deleting the condition are available Click the Stop button to stop the running time lapse experiment Time tab Select to display the capturing interval time total observation time and number of round
66. een For the BS IM MC MOT chamber with the AF mode enabled AF is automatically performed for the currently selected well and the well moves to the X Y position that is previously displayed BioStation IM Version 2 20 Build 140 2 T Time lapse images Temp Chamber 370 570 4 Water 380 380 time lapse setting Acquired Outside 260 unitdeg Stable Y 1 Start C 5 1 1 Ph Sms 20x V 1 2 Ph Sms 20x 2 V 1 3 Ph Sms 20x Register 8 New point DL 414 311 207x156um 104x78um Simple J Manual AE Load settings EPILamp 12 4 4 Exposure time 300ms v Gain 100 4 b Resolution 640 x 480 Binning 7 2 Position 150 00 um Fix af 1 E n n 1 X 163998 VL com Y 396005 3 1 1 24 New time lapse setting screen Live screen Chapter 3 New Time lapse Setting Screen Table 3 1 1 Functions on the New time lapse setting screen Live screen Item Function 1 Select to switch between the Live screen and the Wide field screen buttons 2 Observation The field of view is indicated by the red frame and displayed as the live observation point image verification The registered observation point is indicated by the blue frame display Click the desired point to move the stage and to display the live image of
67. een magnification is fixed to 20x Therefore changing magnification is not available Select to register points in an observation area in blue frame selected on the Wide field screen in a batch Click the Time lapse experiment point registration button with this checkbox checked to register the points in the selected area in blue frame in a batch The points will be registered from left top gt right top gt left bottom gt right bottom When the total of registered observation points and points to be registered exceeds 99 points or selected area overlaps a registered Wide field area that is the XY coordinates match completely the batch registration is not available Wide Field 44 Chapter 3 New Time lapse Setting Screen Table 3 2 4 Operations for setting observation conditions and moving the stage 2 2 Item Function Image display Select magnification of tiled image display with these buttons magnification Full Entire observation area is displayed button 2x mage is enlarged by twice 4x Image is enlarged by four times 20x Displayed image size is one sheet of a tiled image 40x Image is enlarged by 40 times 80x Image is enlarged by 80 times Click each direction button of the jog dial to scroll the tiled image when the entire tiled image is not displayed due to partial enlargement At the center part of the jog dial X and Y coordinates of Fine focus current pointer are displayed
68. en This screen cannot be displayed by operating any buttons Select to playback performed time lapse images This screen automatically appears after time lapse experiment ends On this screen loading and reproducing saved file of time lapse experiment results are available Chapter 1 Basic Operations 1 2 2 Status Display and Settings of the Microscope This area shows the current temperatures and the temperature settings of the culture chamber and the humidifier water tank The outside air temperature is also shown here For the temperature settings of the culture chamber and the humidifier tank see Step 3 in Section 2 1 Start up in the separate manual BioStation IMa Instructions lt System gt Temp chamber 372 375 24 Water 370 z0 0 370 4 untag Stable Unstable Figure 1 2 3 Temperature status display of the microscope Table 1 2 2 Functions on the status display and settings of the microscope Item Function Temp Click the Temp button to display the Temperature dialog box The temperature change since the software is started up and temperature status during time lapse experiment can be checked Chamber Display area Temperature detected with the temperature sensor of the culture chamber is displayed Setting area Set temperature inside the culture chamber to be suitable for time lapse experiment Water Display area Temperature detected with the temperature sensor of the humidifier water tank
69. et the displayed image EM Is saved Figure 4 1 7 Save confirmation dialog box 49 Chapter 4 Time lapse Images in Process Screen 4 1 4 Magnification Button 4145311 Z xi55um 104 7 Oum Figure 4 1 8 Magnification button Table 4 1 5 Magnification function Item Function Magnification button Magnification used for running time lapse experiment is displayed 4 1 5 Z stack Display Control Z stack sequence icon 16 05 Figure 4 1 9 Z stack display Table 4 1 6 Functions on the Z stack display Item Function Z stack The means to capture observation image in the Z direction during the time lapse sequence icon experiment is indicated Pull down The Z coordinate of the image currently displayed is displayed Select the Z coordinate menu of the captured image displayed in the pull down menu to switch the time lapse observation image display Slider Switch the Z coordinate to the positive or negative side from the reference observation point for the image display Z focus mark c Select to register the optimum image displayed by switching the slider bar as button J the focus point Z focus graph button Select to display the Z focus graph dialog This allows you to check the points registered by using the Z focus mark button on a graph 50 Chapter 4 Time lapse Images in Process Screen Graphical display of focus positions Clicking the Z focus graph button E shows the Z fo
70. gs soon as possible 0h 00m 283 Figure 3 1 16 Timelapse dialog box for new registration 31 Chapter 3 New Time lapse Setting Screen Registering a new time lapse experiment time for stream capturing When the conditions for the stream setting are fulfilled the Stream setting box appears on the Timelapse dialog box for new registration After configuring the time lapse experiment time for stream capturing click the Add button to register the configured settings Stream setting limitations The following observation conditions must be applied for the stream setting The Stream setting box does not appear on the Timelapse dialog box unless all of the following conditions are met e The exposure time must be less than a second e One type each for the filter and the objective must be selected e A single observation point Z stack must be applied in the Z direction Timelapse Acquisition cycle 15 minutes Total time minutes Stream blocks 4 Stream setting Stream time 10 gec 4s soon as possible Oh 00m 19s Close Figure 3 1 17 Timelapse dialog box for stream capturing Table 3 1 8 Functions for registering the time lapse experiment time for stream capturing Item Function Acquisition cycle Input the capturing interval time for the time lapse experiment Select a unit from the pull down menu next to the input box Total time Input the total observation time of the time lapse experimen
71. h modes are available for settings in this section x b d 41 4x311 um 207 15 104x78um Simple JC Manual Load settings Lamp Exposure time Gain Resolution Position Figure 2 2 1 Item Filter button Magnification button Simple button Manual button Eoo 1 414 311 207 158um 104x78um AEffocus Simple Manual f Bright Middle f Dark Cy 1003 46 _ EPI Lamp Cy p B ms Exposure time Booms 100 4 _ b Gain E 540 x 480 Binnine Resolution 540 x 480 Binning 150 00 um j m Z Position 150 00 um Fix Setting observation conditions for fluorescent filter Left Manual mode right Simple mode Table 2 2 1 Functions for setting observation conditions 1 2 Function Select the filter for observation The blue frame appears on the selected filter button This operation is also available with the Observation method switches of the ergonomic controller 414x311um 207 15 104x78um Simple Select to perform phase contrast microscopy with the diascopic illumination of the red LED illuminator built in the microscope Fluorescent filter is not used Select to perform the fluorescent microscopy using the selected filter with the illumination of external mercury lamp This button does not appear unless the HG precentered fiber illuminator is connected and turned t
72. h the Shutter x open close switch of the ergonomic controller For phase contrast filter set the intensity of diascopic illumination built in LED illuminator Turn on or off the built in LED illuminator Turned off 21 Chapter 2 Live Observation Screen Table 2 3 1 Functions for setting observation conditions 2 2 Item Function Exposure Select exposure time from the pull down menu time Setting value can be increased or decreased by 1 with the W and A buttons Gain Click the value displayed next to the Gain to display the Edit detailed gain dialog box In the dialog box set the compensation value with the keyboard The compensation value can also be set with the slider and the right and left triangle buttons displayed next to the value Resolution Select resolution from the pull down menu Resolution can be set for each channel separately The applicable combinations are shown as follows Resolution combinations for BioStation IMQ 640x480 640x480 Binning 1280x960 640x480 1280x960 Resolution combinations for BioStation IM Lor rf D ssoxaso pining oc 800x600 Binning v v Y applicable None not applicable 2 3 1 Gain Compensation Value Fine Adjustment Click the compensation value displayed next to Gain to display the Edit detailed gain dialog box Gain compensation value can be adjusted finely Edit detailed gain Figure 2 3 2 Edit deta
73. hannel Select to save the intensity data into a file Multi ch Intensity data This option is enabled only when the intensity is calculated Time lapse log Select to save the time lapse experiment log into a file Select to save the comments of the user mark and cell stimulus marks This option is enabled only when the user mark or the cell stimulus mark is used Mark comment Next button Select to display the Save images Single point dialog box In the dialog box specify the destination to save image and input its file name and file format Back button Select to return to the Save image dialog box Cancel button Select to close the Export images and data Single point dialog box 68 Chapter 4 Time lapse Images in Process Screen Image data save settings for a single observation point 2 2 Click the Next button of the Export images and data Single point dialog box to display the Save images Single point dialog box Save images Single point Falder and Documentss Save image Format oy Gray Color e settings Interval Time fps Total Time 25 zl sec Frequency every 2 frames For Fh Hes sach080512_ch For Hes sach080512_ch2 0 For Fiz test sach080512_ch3 0 For Multi ch lw Save z stack images Filename According focus setting According range setting rom 2050 To 050 r Advanced settings
74. he power on Select the magnification for observation This operation is also available with the Magnification adjustment selector switch UP DOWN of the ergonomic controller Select the Simple mode Set the observation condition selecting from the following three given exposure conditions e Bright to brighten dark images e Middle to observe half bright images e Dark to darken too bright images Set a condition for each combination of filters Ph FI1 and FI2 and magnifications three types Select the Manual mode Set all observation conditions manually or load the already registered condition file Set a condition for each combination of filters Ph FI1 and FI2 and magnifications three types 18 Chapter 2 Live Observation Screen Table 2 2 1 Functions for setting observation conditions 2 2 Item Function Z Position Register an appropriate Z position focus position for each selected filter For example set the Z position for the fluorescent filter 1 F11 and click the Fix button to display the input value in red and register it Even if the fluorescent filter 2 FI2 is selected and the Z position is changed the registered value appears when the fluorescent filter 1 F11 is selected again Click the Fix button again to clear the registered value Additionally changing the Z position is available even after once registering the Z position To display the registered Z position again click the Go button These se
75. igure 4 1 40 Print dialog boxes Table 4 1 19 Functions of captured image printing Item Function Select channel Select the image to be printed For Channels display select a filter For Points display select an observation point The filters that are not specified for the time lapse experiment condition are disabled Print button Select to print an image Cancel button Select to stop printing and close the dialog box 65 Chapter 4 Time lapse Images in Process Screen 4 1 14 Captured Image Saving Click the Save button 7 to display the Save image dialog box If a clipping area is set the save confirmation dialog box for the clipped image appears Click the OK button in the dialog box Then the Save image dialog box appears Export images and data Single point Export images and data Multi points Export large images Multi points Cancel Figure 4 1 41 Save image dialog box Table 4 1 20 Functions of captured image saving Item Save as BioStation IM format button Export images and data Single point button Export images and data Multi points button Export large images Multi points button Cancel button Function During time lapse experiment this button is disabled because the file is already saved at the beginning of the experiment Select to save the time lapse images captured at a single observation point as image file s The Export images and data S
76. iled gain dialog box Table 2 3 2 Functions of gain compensation value fine adjustment Item Function Compensation Set the compensation value with the keyboard or the and A buttons value setting Apply button Select to apply selected compensation value and close the dialog box Close button Select to cancel compensation value setting and close the dialog box 22 Chapter 2 Live Observation Screen 2 4 Observation Point and Focus Adjustment Focus L A Jog dial 4 300033 SEMEN m 350 95 um Y Figure 2 4 1 Setting observation point Table 2 4 1 Functions of observation point movement and focus adjustment Item Function Jog dial Click each direction button of the jog dial to move stage i focus At the center part of the jog dial X and Y coordinates of current stage position are displayed Fine This operation is also available with the X stage knob or the Y stage focus knob of the ergonomic controller Focus button Coarse Click the up arrow button or down arrow button to move objective in focus the Z direction Fine This operation is also available with the Focus knob of the ergonomic focus controller Undo button Select to return observation point to the previous focus position dn lie Slide up or down the slider to adjust focus Well switch Well This button is displayed only for the BS IM MC MOT chamber button Select to switch the well to observe in the glass bottom 4 well dish
77. ime lapse experiment experiment time are displayed Click a time lapse experiment time to display the Timelapse dialog box In the dialog box changing and deleting capturing interval time setting and total observation time setting are available Delete button Select to delete the selected time lapse experiment time The Delete confirmation dialog box appears To delete the selected time lapse experiment time A E 0 click the OK button To return to the Time tab without deleting click the Cancel button However currently running and already done Figure 4 1 35 Delete confirmation time lapse experiment times cannot be deleted as dialog box the Delete button becomes disabled BioStation Red dot Indicates the time lapse experiment is running New button Select to set new time lapse experiment time The Timelapse dialog box appears for new registration of time lapse experiment time Details on the Timelapse dialog box EE Operation of the Timelapse dialog box is the same as that of the New time lapse setting screen hours s soon as possible 0h 00m 28s Reference Chapter 3 New Time lapse Setting Screen Delete Section 3 1 4 Time lapse Experiment Scheme Time Tab Changing or deleting time lapse experiment time Registering new time lapse experiment time EE minutes minutes s soon as possible Oh 00m 28s Figure 4 1 37 For new registration 62 Chapter 4 Time lapse
78. ingle point dialog box appears Select to save the time lapse images captured at multiple observation points as image files The Export images and data Multi points dialog box appears Select to save the time lapse images Wide field image as image file s The Export large images Multi points dialog box appears Select to close the Save image dialog box 66 Chapter 4 Time lapse Images in Process Screen Image data save settings for a single observation point 1 2 Click the Export images and data Single point button of the Save image dialog box to display the Export images and data Single point dialog box Export images and data Single point Export images and data of Paint 1 and 205 Save time range t AJ One shot of current time Neighbors of current time Time 0 00 00 ta 24758 Save channel Ph use pseudo color iw use pseudo color Time lapse log n zu Figure 4 1 42 Export images and data Single point dialog box Table 4 1 21 Functions of image data save settings for a single observation point 1 2 Item Function Export images and Select the observation point and magnification to save the image from the pull down data of menu Save time range Select to save images of the selected observation point under the selected magnification in a file One shot of current time Select to save only the currently displayed
79. just the intensity and gain until there is no saturation point BioStation IM Version 2 20 Build 140 Temp Chamber 370 370 Water 380 80 EA mw Time lapse images Time Peel images i i 21 Water m time mw setting in process Acquired Outside 260 uitdeg Stable gt 3000um uz 414 311 207 15 104x78um Simple Manual AEfoous AE Load settings EPI Lamp 100 4 Exposure time 300ms X j Gain 100 4 Resolution 640 x 480 Binning Z Position 150 00 um Well A 1 A m X 300033 4 a 4 Figure 2 1 5 Displaying saturation point 15 Chapter 2 Live Observation Screen 2 1 4 Intensity Screening Click the Intensity screening button ri to display the Intensity Screening dialog box Intensity screening Apply Close Figure 2 1 6 Intensity screening dialog box Table 2 1 4 Intensity screening functions Item Function Enable Screening checkbox Select to enable the intensity screening functions Image select box Drag a marquee left click and drag around the area for intensity screening yellow box Enlarge reduce or move the area with a mouse pointer if necessary Right click the box to display the Delete menu to delete the box Image display Screening points same intensity points are displayed in blue Histogram Inten
80. k the Cancel button Figure 3 1 12 Delete all confirmation dialog box Load button Select to load the registered time lapse experiment condition file Save button Select to save the registered observation point observation condition and time lapse experiment time to a time lapse experiment condition file Registering multiple time lapse experiment times is available Acquisition cycle Totaltime Rounds This software can register multiple time lapse experiment times wong 9 Oh00m 10s Oh 10m 00s 61 which are composed of different total observation time and Oh 10m 00 20h 00m 0068 121 Delete capturing interval time When a time lapse experiment starts the multiple time lapse experiment times registered with the Time tab are automatically C e applied in descending order Figure 3 1 13 Time tab Additionally even during the time lapse experiment adding a new time lapse experiment time and changing or deleting unperformed time lapse experiment time are available 30 Chapter 3 New Time lapse Setting Screen Changing or deleting time lapse experiment time Click a time lapse experiment time on the Time tab to display the Timelapse dialog box for setting change Timelapse Acquisition cycle 1 0 minutes Tatal time 20 hours Rounds 121 As soon as possible 0h 00m 28s Apply Delete Figure 3 1 14 Timelapse dialog box for setting change Table 3 1 7 Functions for changing
81. lder dialog box and specify the destination to save C My Documents 4 Computer 8 314 Floppy Sw Local Disk C D Bos COMPAQ C Documents and Settings Drivers 3i hp x 23 i386 c Image New Folder Browse For Folder Make New Folder Figure 4 1 49 Browse For Folder dialog box Save image Format Select a format from six formats to save images TIFF Gray 16 bits Color 24 bits TIFF Gray 8 bits Color 24 bits BMP Gray 8 bits Color 24 bits PNG Gray 8 bits Color 24 bits JPEG Gray 8 bits Color 24 bits AVI 75 Chapter 4 Time lapse Images in Process Screen Table 4 1 25 Functions of image data save settings for multiple observation points 2 2 Item Function Save image AVI setting Enabled when AVI is selected in the Format field Interval Time opecifies the frame rate number of frames displayed per second at which pictures taken are saved into an AVI file maximum frames 60 Total Time opecifies the time during which pictures taken are saved into an AVI file The number of frames and total time are calculated and if the frame rate is above 33 fps an error is indicated Frequency every Enabled when this checkbox is enabled Allowable range 2 to 99 The thinning out setting is valid for both Interval Time Total Time Filename Input a file name to save images Channel names are added to the end of file names automatically Ph ch1 Fl1 ch2 FI
82. lective applicable aux pays europ ens Ce symbole indique que ce produit doit tre collect s par ment Les mesures suivantes concernent uniquement les utilisateurs euro p ens e Ce produit doit tre jet s par ment dans un point de collecte appro pri Ne jetez pas ce produit dans une poubelle r serv e aux ordures m nag res Pour plus d information contactez le d taillant ou les autorit s locales responsables de la gestion des ordures Es S mbolo para recogida separada aplicable en pa ses Europeos Este s mbolo indica que este producto se recoger por separado Lo siguiente s lo se aplicar en pa ses Europeos Este producto ha sido designado para su recogida en un punto de al macenamiento apropiado No lo tire como un deshecho dom stico Para m s informaci n contacte con el vendedor o autoridades locales al cargo de la gesti n de residuos Dki Symbol for special bortskaffelse af denne type produkter i de europ iske lande Dette symbol angiver at dette produkt skal bortskaffes specielt Det efterf lgende er kun til forbrugere i de europ iske lande Dette produkt skal bortskaffes p fx en genbrugsplads el lign Det m ikke smides v k som normalt husholdningsaffald For yderligere information kontakt din forhandler eller de lokale myndigheder som fx teknisk forvaltning Symbool voor gescheiden inzameling zoals dat wordt gebruikt in Europese landen Dit symbool betekent dat dit prod
83. mage display is marked with a blue box Only the image at the checked observation point is displayed on the observation image display Up to four observation points can be displayed at a time Click this button to calculate intensity value If this button is clicked after an area of an observation image is selected the intensity graph for the selected area is displayed The ROI mark is normally marked with a yellow frame and the ROI mark when intensity graph is displayed is marked with a red frame 88 Enl Symbol for separate collection applicable in European countries This symbol indicates that this product is to be collected separately The following apply only to users in European countries This product is designated for separate collection at an appropriate collection point Do not dispose of as household waste For more information contact the retailer or the local authorities in charge of waste management Del Symbol f r getrennte Wertstoff Schadstoffsammlung in europ ischen L ndern Dieses Symbol zeigt an dass dieses Produkt separat entsorgt werden muss Folgendes gilt f r Verbraucher in europ ischen L ndern e Dieses Produkt darf nur separat an einer geeigneten Sammelstelle entsorgt werden Eine Entsorgung im Hausm ll ist unzul ssig e Wenden Sie sich f r n here Informationen bitte an Ihren H ndler oder die rtlich f r Abfallentsorgung zust ndigen Beh rden 154 Fr Symbole pour la collecte s
84. ments of the film bottom 4 quadrant dish on the Cell name etc tab Cell name etc Sample name Cell name User name Dimension First quadrant test test2 test Second quadrant test2 Third quadrant test3 Fourth quadrant test4 test3 test4 Figure 1 3 11 Cell name etc tab BS IM Enable AF Mode Select to enable the AF mode and display the Reference mark register MC MOT screen Auto focusing is carried out whenever you switch between the chamber wells on the New time lapse setting screen Launch as Mark Select to display the Reference mark register screen immediately after window mode the software is started If this is not selected the Live observation screen appears instead 125 5 Hide Control Panel Click the Hide control panel menu to display only the image view on the left side of the window BioStation IM Version 2 20 Build 140 Live observation Figure 1 3 12 Hide control panel Live Observation Screen When the BioStation IM IMQ is used as a microscope select this screen You can check the specimen is suitable or not for time lapse experiment by observing cell shape and the amount of fluorophore in cell 1 2 3 4 5 6 BioStation IM Version 2 20 Build 140 time lapse setting Time lapse images Temp Chamber 370 370 Water 380 980 H Acquired Outside 260 unitdeg C Stable 2
85. mouse pointer on the button enlarged image When the image is dragged without clicking the scroll mode button or clipping button a region of interest ROI area is drawn 4 1 3 Setting a Clipping Range of the Displayed Image Figure 4 1 5 Clipping button Table 4 1 4 Function for setting a clipping range of the displayed image Item Function Clipping Select to set a clipping range of the displayed image button Select left click and drag an area of the displayed image A clipping range of the image can be set If this operation is performed for one image the same clipping area is set for other three images However when a clipping area is set for an image only the set clipping area clipping image can be saved Click the clipping button again to turn off the clipping Bio Station IM function To delete the set clipping area display the delete confirmation pee ibang dialog box by right clicking the clipping area and selecting Delete from the submenu or selecting the clipping area EM and pressing the Delete key on the keyboard then click the Figure 4 1 6 Delete confirmation OK button in the dialog box dialog box Saving an image when a clipping area is set with the clipping function Bio Station IM If a clipping area is set click the Save button to display the save giu IM LI confirmation dialog box for the clipping image and click the OK button to save the clipping image If a clipping area is not s
86. mple name Cell name and User name are displayed Click any displayed name to display the Cell name etc dialog box In the dialog box inputting changing and deleting the name are available The time lapse experiment can be performed even if none of them is input The input information in the Cell name etc dialog box can be displayed on saved images or Live images This field appears when Hi Q4 film bottom 4 quadrant dish is set on the Preferences dialog box Since the BS IM MC MOT chamber does not support the Hi Q4 film bottom 4 quadrant dish the Dimension field does not appear See Section 1 3 4 Basic Settings as a reference Click the Dimension field to display the Dimension dialog box In the dialog box inputting changing and deleting the names of the first to fourth quadrants are available Select to load the saved time lapse experiment condition file Select to save the registered observation point observation condition and time lapse experiment time as a time lapse experiment condition file 33 Chapter 3 New Time lapse Setting Screen Inputting subordinate information of time lapse experiment Click Sample name Cell name User name on the Cell name etc tab to display the Cell name etc dialog box Cell name etc Sample name Cell name User name Apply Figure 3 1 19 Cell name etc dialog box Table 3 1 10 Functions for inputting time lapse experiment subordinate information Item F
87. n 7 All highlighted images are saved at the same time Click the Trash button to delete the selected thumbnail images Observation condition and observation point Adjust the stage position the focusing condition and the observation conditions such as filters magnification light intensity and exposure time 10 Chapter 2 Live Observation Screen 2 1 Image Adjustment Buttons and Captured Image Display Figure 2 1 1 Image adjustment buttons and captured image display Table 2 1 1 Functions of the image adjustment buttons and captured image display 1 2 Item Function Automatic tone curve adjustment Select to automatically set the range Display range of intensity is apply button automatically set The highest intensity A of the displayed image is set as the upper limit C and the lowest intensity B is set as the lower limit D D A O Figure 2 1 2 Intensity graph Select to set tone curve adjustment manually adjustment setting button The Tone curve adjustment dialog box appears to set tone curve adjustment Tone curve adjustment cancel button Select to cancel the applied tone curve adjustment and return to the original screen FL2 only To clear the saturation check condition clicked this button again This function can be used only for the fluorescent microscopy Intensity screening button FL1 amp FL2 only Pseudo color
88. n Select to return to the Export large images Multi points dialog box Cancel button Select to close the Save large images Multi points dialog box 80 Chapter 4 Time lapse Images in Process Screen 4 1 15 Example of a Saved Image Observation conditions Input information on the Cell name etc tab Filter Ph Sample name Sample 01 Cell name Cell 01 User name User Dimension Divi Fassage of tine Uh Om 37s Figure 4 1 53 Example of a saved image When the 4 quadrant dish is selected Scale Displaying the number of rounds photo date and passage of photo time 81 Chapter 4 Time lapse Images in Process Screen 4 2 Points Display This screen shows process of time lapse experiment One display area is provided for one observation point Switch the display area to display each filter image and overlapped image Therefore up to four observation points can be observed at the same time Functions other than the way to display images are the same as those for the Time lapse images in process Screen Channels Bio Station IM Version 2 12 Build 105 test_sach08051 2 apse images Temp chamber 37 0 70 Water 378 380 in process Outside 293 unitideg C Stable Qe ae W EJ g 1 3000 33 4459 00 1 Ph Et 2 Ratio Y 414 311 207x156um iM Bum __ 1 2 File name test_sach080512 2 2 Point
89. ng and deleting time lapse experiment time setting are available Click the New button to add new time lapse experiment time Cell name Select to input subordinate information of time lapse experiment etc tab Sample name Cell name or User name can be input Click any displayed name to display the Cell name etc dialog box In the dialog box inputting changing and deleting the name are available Zstack tab To specify multiple observation points in the Z direction for an Observation point use this function Up to 99 observation points can be set by specifying the travel amount and travel count along Z axis However the setting that exceeds the maximum stroke in the Z direction is not available 7 Observation Set observation conditions filter selection magnification selection intensity condition and adjustment and exposure time setting and move stage and perform focusing with the observation Jog dial point For the BS IM MC MOT chamber the Well switch button is displayed Clicking the Well switch button displays the live observation image of the selected well 8 Time lapse After specifying all observation conditions click the Time lapse experiment experiment point registration button to register the specified conditions point to the Time lapse experiment scheme registration If registered observation condition setting is changed the setting is button overwritten Up to 99 points can
90. ning time lapse experiment 63 Chapter 4 Time lapse Images in Process Screen Displaying the logarithmic graph of temperature change Click the Temperature log button v in the time lapse experiment process area to display the following dialog box Temperature logging start at Tuesday July 15 2008 14 35 11 E The time lapse experiment is running under a stable condition Time lapse running Temp Fluctuations 2 48 lt 3himizs Zh1m3B8s Close An unstable condition is detected during the time lapse experiment Figure 4 1 39 Temperature dialog box Table 4 1 18 Functions on the Temperature dialog box Item Function Range Select time range horizontal axis of temperature graph Temperature graph Temperature changes inside of the culture chamber and the humidifier water tank and that of ambient temperature are shown Close button Select to close the Temperature dialog box 64 Chapter 4 Time lapse Images in Process Screen 4 1 13 Captured Image Printing Click the Print button to display the Print dialog box The Print dialog box changes depending on the observation image conditions Channels display or Points display Use this dialog box to print the images captured with time lapse experiment Select channel lw Point 2 lw Point 3 Iw Multi ch lw Point 4 v Intensity Graph wv Intensity Graph Caneel Caneel Print dialog box for Channels display Print dialog box for Points display F
91. oint is indicated by the blue frame 5 Image adjustment Select to set up tone curve adjustment buttons Operation of these buttons is the same as that of the Live observation screen 6 Partial enlargement Click the Magnifier button and drag the mouse pointer on the tiled image to enlarge the area according to the magnification selected from the pull down menu 7 Cross line display The Show cross line checkbox appears when Hi Q4 film bottom 4 quadrant checkbox dish is selected on the Preferences dialog box Select to display the crosshair This software identifies the quadrants 1 to 4 with the crosshair 39 Chapter 3 New Time lapse Setting Screen Table 3 2 1 Functions on the New time lapse setting screen Wide field screen 2 2 Item Function 8 Start time lapse Select to start a time lapse experiment button A time lapse start confirmation dialog box Confirmation window appears 9 ime 9 Time lapse Displays and functions of the tabs are same as those of the Live screen experiment scheme 10 Observation condition Switch filters and a magnification to be used ae point When an enlarged view of the tiled image is displayed the display area can settings be scrolled with the operation of these controls 11 Save Wide field image Select to combine range selected observation area and save it as one Wide field image Click this button to show the Save As dialog box Select the im
92. on conditions Save as Simple Setting for Item Function Save as Simple Select to save the observation condition as the setting file in the Simple mode Setting for Specify the exposure condition for which the observation condition is saved Save button Select to display the save confirmation oe dialog box To save setting click the OK button To return to the Save settings dialog box without saving click the Cancel button Figure 2 2 4 Save confirmation dialog box Close button Select to close the dialog box without saving observation condition setting 20 Chapter 2 Live Observation Screen 2 3 Setting Observation Conditions Focus Mode Automatic Exposure Condition File Loading Light Intensity Exposure Time Gain and Resolution Each setting in this section is available only on the Manual mode 2 P d 207x156um 104x78um 414 311um 207 156um 104x78um Sinis Load settings Simple Manual Load settings AE AE focus EPILamp Q 100 4 DALam 71 4 Exposure time B ms Exposure time 14496 35 Gain P Gain 16 00 4 1 bh Resolution 1640 x 480 Binning Resolution 640 x 480 Binning v Z Position Z Position 350 95 um 150 00 um Fix Figure 2 3 1 Setting observation conditions on the Manual
93. on image display is marked with a blue box Only the image at the checked observation point is displayed on the observation image display Up to four observation points can be displayed at a time Select to calculate intensity value If this button is clicked after an area of an observation image is selected the intensity graph for the selected area is displayed The ROI mark is normally marked with a yellow frame and the mark when an intensity graph is displayed is marked with a red frame 82 Time lapse Images Acquired Screen This screen automatically appears at the end of time lapse experiment On this screen loading and reproducing the saved time lapse result files are available There are the Channels display and the Points display for the Time lapse images Acquired screen A part of functions is different from those of the Time lapse images in process screen This section indicates only the different part File compatibility of the saved file of time lapse result files Files created by the software of Ver 1 to Ver 2 0 can be read into the software of Ver 2 1 or later whereas those created by the software of Ver 2 1 or later cannot be read into the software of Ver 1 Additionally if a file created by the software of Ver 1 to Ver 2 0 is read into the software of Ver 2 1 or later it is automatically upgraded to the file format of the version currently used Once upgraded it cannot be read into the sof
94. ons Explorer launches Select to display the Explorer after captured images are saved when image capture Explorer launches Select to display the Explorer after images are exported in the general when file export format Viewing Exposure Time Select the unit of exposure time displayed on the software msec Fractional Fractional representation in seconds Only the Fractional representation is available when BioStation IM is used Load settings DIALamp 0 95 4 Load settings DALam 55 4 Exposure time 1 250s Exposure time Gain Gain Resolution Resolution Z Position Z Position Figure 1 3 7 representation Figure 1 3 8 Fractional representation Chapter 1 Basic Operations Table 1 3 2 Functions of Preferences dialog boxes 2 2 Item Function Viewing EPI Lamp Select the unit of the light intensity of the EPI lamp displayed on the software Transmission rate Position representation Only the Position representation is available when BioStation IM is used Load settings Load settings EPILamp Q 25 4 4 EPllam 4 4 Figure 1 3 9 Transmission rate Figure 1 3 10 Position representation representation Dish Type Select the dish type Normal Hi Q4 used for observation Selecting Hi Q4 shows the fields to input and edit the names of the compart
95. or S alid Lum 120 Blue 0 Cancel Add to Custom Colors Figure 4 1 15 Color dialog box Apply button Select to close the dialog box applying the settings of file name and color Close button Select to close the dialog box without applying settings o4 Chapter 4 Time lapse Images in Process Screen Setting ratio Click the Ratio setting button Fna ofthe time lapse experiment image Multi Ch to display the Ratio Setting dialog box Ratio Setting Ratio Channel FIT FI2 Ratio Range 0 25 4 00 Fit Background Threshold M ea 2 Apply Close In this area set the lower limit of the intensity for ratio setting Figure 4 1 16 Ratio Setting dialog box Table 4 1 11 Functions for setting ratio Item Function Ratio Channel Select Ratio Channel from the pull down menu Ratio Range Input Ratio Range Apply button Select to register Ratio settings Close button Select to close the dialog box without applying Ratio settings 55 Chapter 4 Time lapse Images in Process Screen 4 1 7 Intensity Graph Time Range Ft Intensity Range FI1214 2 94 Ratio 0 1489 Calculate Intensity 2398 1798 1199 Item Time Range Intensity Range Grid button Calculate button Oh 00m 38s Oh Odm 16s Oh 1m 54s Oh zm 31s Oh 03m 09s Oh 3m 47s Figure 4 1 17 Intensity graph Table 4 1 12 Functions of intensity graph Function Select the time range horizontal axis of one page graph from the
96. or deleting time lapse experiment time Item Function Acquisition cycle Input capturing interval time for time lapse experiment Select a unit from the pull down menu next to the input box Total time Input total observation time of time lapse experiment Select a unit from the pull down menu next to the input box Rounds Round value is displayed This value is automatically calculated from total observation time and capturing interval time Apply button Select to apply the input values to time lapse experiment time setting Delete button select to delete the highlighted time lapse BioStation IM experiment time The Delete confirmation dialog box appears To delete the selected time lapse experiment time click the OK button To return to the Timelapse dialog box without deleting click the Cancel button Figure 3 1 15 Delete confirmation dialog box Close button Select to cancel the Time lapse experiment time setting and close the dialog box Registering new time lapse experiment time Click the New button of the Time tab to display the Timelapse dialog box for new registration Setting values of capturing interval time and total observation time are the same as those of the Timelapse dialog box for setting change After setting time lapse experiment time click the Add button to register the setting Timelapse Acquisition cycle 1 minutes Total time 2 minute Rounds 2 A
97. owing checkboxes to specify the font size and color of the characters for the image to be written opecifies font of the character written into the image opecifies size of the character written into the image opecifies color of the character written into the image settings written into the image to the OK button Select to save the settings and to return to the Save images Single point dialog Cancel button Select to close the Save images Advanced settings dialog box 72 Chapter 4 Time lapse Images in Process Screen Image data save settings for multiple observation points 1 2 Click the Export images and data Multi points button of the Save image dialog box to display the Export images and data Multi points dialog box Export images and data Multi points Select export data Select ALL Points v Point 1 v Point 2 v Point 3 Point 4 Save lime range C Al One shot af current time Neighbors of current time C Round m to E Time 2 00 00 ta 24758 Save channel v Ph lw use pseudo color iw Fiz use pseudo calor e Figure 4 1 47 Export images and data Multi points dialog box Table 4 1 24 Functions of image data save settings for multiple observation points 1 2 Item Function Select export data Select ALL Points Select to highlight all observation points Point 1 to Point n All registered observation points appear here To select
98. p button Select to stop the running time lapse experiment AN Stop ol The stop confirmation dialog box appears To stop time lapse experiment click the OK cancel button Figure 4 1 32 Stop confirmation dialog box Resume button When time lapse experiment is stopped the Stop button is changed to the Resume button Click the Resume button to restart time lapse experiment Checking observation condition Only checking the observation condition is available in the Point information dialog box displayed this time 2 414x311 um DIA Lamp 150 Exposure time 5ms Gain 1 00 Resolution 640 x 480 Binning Dish 134419 um 5021 86 um 150 00 um 14850um 151 50um steps Comment test Figure 4 1 33 Point information dialog box Table 4 1 15 Function for checking the observation condition Item Function Close button Select to close the Point information dialog box 61 Chapter 4 Time lapse Images in Process Screen 4 1 11 Time lapse Experiment Scheme Time Tab When the stream setting is configured during the time lapse experiment time setting the display of Time tab is replaced with Stream Acquisition cycle Totaltime Hounds Oh 2m 00 10h 00m 00s 31 OhOOm 10s Oh 10rn 00 1 Oh 10m 00 20h 00m 00s 121 Delete New Figure 4 1 34 Time tab Table 4 1 16 Functions on the Time tab Item Function Time lapse Capturing interval time total observation time and rounds for t
99. played image in a file Neighbors of Select to save the currently displayed image the five current time preceding images and the five following images total of 11 images into a file Concatenate Select to save all concatenated images into a file all marks The concatenated images consist of images of all marks including user marks and cell stimulate marks added in the Time Line five preceding images of each mark and five following images of each mark If neither user mark nor cell stimulate mark is set this item is disabled Input time range Saves the image in the specified round range as a file Round According to the round count specified in these fields the time of the Time field is automatically set Saves the image within the specified shooting time as a file According to the time specified in these fields the count of the Round field is automatically set Time Select to save the Ph channel data into a file 11 Select to save the Fl1 channel data into a file To save the image with the pseudo color check the use pseudo color checkbox Select to save the 2 channel data into a file To save the image with the pseudo color check the use pseudo color checkbox Multi ch Select to save overlaid image into a file with the displayed condition Next button Select to display the Save large images Multi points dialog box In the dialog box specify the destination to save image and input its file name
100. pull down menu Fit total observation time fits in one page graph 1x 2x 3x and 4x calculated from minimum interval time value of Time Range and coefficient Time per scale 4 value of Time Range x minimum interval time Time range of one page graph time per scale x 7 Example when value of Time Range is 2x and minimum interval time is 10 minutes 4 2 x 10 20 minutes time per scale 20 x 7 140 minutes time range of one page graph If total observation time cannot fit in one page scroll bar appears at the bottom of the graph Select the intensity range vertical axis of one page graph from the pull down menu Fit full range of measured intensity fits in one page graph 1x 2x 3x and 4x calculated from the following equations when one unit is the range of to the maximum intensity measurement result 4 Intensity range of one page graph 4 value of Intensity Range x one unit Example when value of Intensity Range is 2x and one unit is 600 Intensity O to 2400 4 2 x 600 1200 intensity range for one page graph When value of Intensity Range is 2x or more a scroll bar appears at the right side of the graph Select to display or hide grid line on graph Select to calculate intensity value If this button is clicked after an area of an observation image is selected the intensity graph for the selected area is displayed See Section 4 1 8 Intensity Graph Displaying Method 56 Chapter 4
101. r using BioStation IMQ screens as examples Procedures are virtually identical for BioStation IM series e This manual describes various operations in Windows XP Vista 7 using Windows XP screens as examples Procedures are virtually identical for Windows XP Vista and 7 Depending on the specific OS type or version the actual appearance of the screen or operations may not correspond precisely to the example screens shown at various points throughout the manual For information on operations or screens specific to your OS version please consult the OS user s manual Contents um TR un i Notes on Handling the SoftWare a aaa E a li Chapter 1 Basic Operations viai oa usc DU ea e siue Ne Vd 1 1 1 Starting and Closing the 1 1911 Slamming tie SOItWaEGe ake deo Ie 1 1 1 2 Registering AF Position of the Reference Mark BS IM MC MOT Chamber 2 11593 SCIOSING the SOMWA E es idees adde 3 50 COMMON FUN OM E OE a 4 12 7 Screen SWwIICIBUllons uos cone ee e IEEE DRE IE 4 1 2 2 Status Display and Settings of the Microscope 5 1 3 Context Menu on the Title 6 1 3 1 Checking the Software Versions
102. ration button to register the AF positions of the reference marks For details see Section 7 4 Setting up the BS IM MC MOT chamber in the separate manual BioStation IMQ Instructions System Chapter 1 Basic Operations 1 1 3 Closing the Software 1 Click the Close button in upper right corner of the window _ ___ 0 Temp Chamber 370 70 Water 378 380 H This software closes and the Windows desktop Outside 293 Stable appears m d Perform this procedure when you detach the Figure 1 1 5 Close button chamber For the BS IM MC MOT chamber this procedure triggers the dish mounting part take the position in readiness to sustain dismounting of the chamber Close confirmation dialog box If the Close button is clicked while captured images remain unsaved the Close confirmation dialog box appears AN Not saved images remains Clase Bio Station IM Bio Station IM To save the captured image click the No button to return to the Bio Station IM screen To close the software without saving the captured Figure 1 1 6 Close confirmation dialog box image click the Yes button Chapter 1 Basic Operations 1 2 Function The items shown on the top screen of the software are common functions for all screens BioStation IM Version 2 20 Build 140 Screen switch buttons Time lapse images Time lapse images Temp
103. s V 1 Ph 142505 FIT 1 45 FI2 1 4s 10x 2 Ph 1 2508 FIT 1 45 FI2 1 48 10x 3 BUT 3 Ph1 250s FIl 1 45 FI2 1 48 10x VE 4 Ph1 250s F1 1 45 FI2 1 45 10x 5 Ph 1 250s 11 1 4s FI2 1 4s 10x 15 05 42 PM Nov 15 05 52 PM 5 63 Time ine BEBE __ Number of rounds 12 61 Time Range Fit v Intensity Range Fit v Grid Calculate 4 Intensity Passage of time Oh 04m 26s Oh 10m 00s 4096 3072 Now captured Point 20x Temperature log ev Oh Q0 m 40s Oh O1m 20s Oh 02m 005 Oh 02m 39s Oh O3m 195 Oh O3m 59s Pause Stop Stream block 2 Round gt b gt EX Oh 00m 005 cm Figure 4 2 1 Time lapse images in process screen Points display Table 4 2 1 Functions on the Time lapse images in process screen Points display Item Function Overlapping buttons Observation image display Selecting observation points Calculate button Select to highlight filters for references The selected filter is checked and the image is displayed Multiple filters can be selected When multiple filters are selected the selected filters overlapped image is displayed Images at different points can be displayed on the four observation image displays Additionally the ROI for intensity analysis can be set for each observation point Click in the observation image display to select an observation point The point number at the upper left of the observati
104. s 1 2 Item Function Image Select to save the image that underwent tone curve Apply tone curve processing adjustment 9 Not available when saving Wide field image Write current ROI niai to write the ROI mark into images for the intensity Not available when saving Wide field image Select to add a color bar to an image for which the ratio is set Add ratio color bar The color bar appears on the right side of the image Not available when saving a Wide field image Overlay text Check this checkbox to enable the following checkboxes to specify the write position for each item to be written into the image For the image in which the information is written see Section 4 1 15 Example of a Saved Image Select to write the observation conditions such as the Observation settings exposure time gain and resolution and select their position of display Select to write the scale and user mark and select their Scale amp User Mark position of display Not available when saving a Wide field image Select to write the cell name and etc and select their position Cell name etc of display Select to write the number of round capturing date and Round information passage of capturing time and select their position of display 71 Chapter 4 Time lapse Images in Process Screen Table 4 1 23 Settings for writing information into images 2 2 Item Function Use font setting Select to enable the foll
105. s for time lapse experiment Click time lapse experiment time display to display the Timelapse dialog box In the dialog box changing and deleting time lapse experiment time setting are available Click the New button to add new time lapse experiment time 47 Chapter 4 Time lapse Images in Process Screen Table 4 1 1 Functions on the Time lapse images in process screen Channels display 3 3 Item Function 14 Time lapse Cell name Select to input subordinate information of time lapse experiment scheme etc tab experiment Sample Cell name or User name can be input as subordinate information Click any displayed name to display the Cell name etc dialog box In the dialog box inputting changing and deleting the name are available 15 Time lapse The time lapse experiment time start and estimated end time of time lapse experiment process experiment number of rounds elapsed time and logarithmic graph of display temperature change are displayed 16 Saturation alert icon If saturation occurs during the time lapse experiment this icon appears Determined as saturation when adjacent two pixels or more reached the maximum intensity continuously Click the icon to display a saturation information dialog This icon once appeared remains displayed after the saturation has been reset 17 Select to print the image captured with time lapse experiment The Print dialog box
106. se button Edit comment button Point information 414x311 um DIA Lamp Exposure time 10 Resolution 640 480 Binning Dish ium c BOOST um BOOST um 0000 0 A5000 um test Delete Figure 3 1 7 Point information dialog box Table 3 1 5 Functions for checking changing or deleting observation condition Function Select to change the displayed settings To change displayed observation condition settings to new settings click the Go button Select to delete the selected observation point The Delete confirmation dialog box appears A Delete from point list To delete the selected observation point click the OK button To return to the Point information dialog box without deleting click the Cancel button Figure 3 1 8 Delete confirmation dialog box BioStation IM Select to close the Point information dialog box Select to add or edit a comment per observation point Click the Edit comment button to display Edit comment dialog box for adding or editing comments Edit comment Click the Apply button to add the comment after creating it canine Click the Close button without test clicking the Apply button to ignore the entered comment and close the Edit comment dialog box Added comment pops up when you double click the observation point Figure 3 1 9 Edit comment dialog box 29 Chapter 3 New Time lapse Setting Screen 3 1 4 Time laps
107. served Click an observation point to display the Point information dialog box In the dialog box checking changing and deleting the condition are available Select to delete the selected observation BioStation point The Delete confirmation dialog box A eese appears To delete the selected observation point click the OK button l To return to the Points tab without deleting click the Cancel button Figure 3 1 5 Delete confirmation dialog box select to delete all selected observation oorr A points The Delete all confirmation dialog box A Delete all points appears To delete all selected observation points click the OK button To return to the Points tab without deleting click the Cancel button Figure 3 1 6 Delete all confirmation dialog box Select to load the registered time lapse experiment condition file Select to save the registered observation point observation condition and time lapse experiment time to a time lapse experiment condition file i If time lapse experiment time setting is disabled or hard disk space of control PC is insufficient this mark appears 28 Chapter 3 New Time lapse Setting Screen Checking or deleting observation condition Click an observation point on the Point tab to display the Point information dialog box In the dialog box check observation condition setting for each combination of filters and magnifications Item Go button Delete button Clo
108. setting the ROI mark m 7 ex Owe e Ph v v 2 Ratio 2 u 414 311 207x156um 104x78um 141 152 1 m LI i m ES Time Range Fit v Intensity Range Ft Ca ir 3072 2048 1024 Oh 00m 36s Oh 01m 12s Oh 01m 48s Oh 02m 23s Oh 02m 58s Oh 03 35s Figure 4 1 21 Selecting the area of image to display an intensity graph e Inthe scroll mode or the clipping mode no ROI mark can be drawn on the image area Therefore cancel the scroll mode or the clipping mode to draw an ROI mark Select the observation point to display the intensity graph Click an observation point of an image to display its intensity graph A blue frame appears on the point number of the selected observation image Display the intensity graph Click the Calculate button The intensity graph of the selected area is displayed in the intensity graph display area When the intensity graph of a selected area is displayed its frame color changes from yellow to red The frame color of other selected areas remains yellow Display area of an intensity graph For information on the time horizontal axis and the intensity vertical axis of the intensity graph see Section 4 1 7 Intensity Graph 58 CO e E TPE Figure 4 1 22 Selecting the area of image to display the intensity graph ELJOO 9 ZANEN TI 2 Fl2 472 Ratio 0 3008
109. sity histogram of the image is displayed To adjust the intensity range drag the and A buttons from side to side Apply button Select to apply setting of intensity screening to the Live observation image display Close button Select to close the Intensity Screening dialog box 16 Chapter 2 Live Observation Screen 2 1 5 Pseudo Color Display Click the Pseudo color button to display the live observation image with the pseudo color of the fluorescent filter You can capture the live observation image displayed with the pseudo color and save it into a file BioStation IM Version 2 20 Build 140 New Time lapse images Time lapse images Temp Chamber 370 370 Water 380 80 H time lapse setting in process Acquired Outside 260 unitdeg Stable 3000um bu 414 311 207x156um 104x78um Simple AE Load settings EPI Lamp 100 4 Exposure time 800ms M Gain 100 4 Resolution 640 x 480 Binning Z Position 15000 um Well X 300033 300033 moe Figure 2 1 7 Pseudo color display 17 Chapter 2 Live Observation Screen 2 2 Setting Observation Conditions Filter Magnification Mode Z position and Save Some observation condition settings vary between the phase contrast microscopy and the fluorescent microscopy Also there are the Manual mode and the Simple mode for the observation condition Bot
110. sult file 86 Chapter5 Time lapse Images Acquired Screen Table 5 1 3 Functions on the Save into Bio Station IM format dialog box 2 2 Item Function Save time range All Select to save all images of the selected observation point under the selected magnification into a file Save points Save tone curve processing parameter Save ROI information Save button Back button Cancel button One shot of current time Select to save only the currently displayed image in a file Neighbors of current time Select to save the currently displayed image the five preceding images and the five following images total of 11 images into a file Concatenate all marks Select to save all concatenated images into a file The concatenated images consist of images of all marks including user marks and cell stimulate marks added in the Time Line five preceding images of each mark and five following images of each mark If neither user mark nor cell stimulate mark is set this item is disabled Input time range Saves the image in the specified round range as a file According to the round count specified in these fields the time of the Time field is automatically set Saves the image within the specified shooting time as a file According to the time specified in these fields the count of the Round field is automatically set Select an observation point to save Select to save the tone curve adjustment parameter Only
111. t Select a unit from the pull down menu next to the input box Stream blocks The number of stream blocks is displayed The value is automatically calculated from the streaming time total observation time and capturing interval time The unit of stream data captured with the stream setting is called the Stream block in this manual Stream setting Select to enable the stream setting Stream time Set the stream capturing time setting range 1 to 3600 sec Add button Select to configure the set values as the time lapse experiment time Close button Select to cancel the Time lapse experiment time setting and to close the dialog box Caution on stream capturing e After the time lapse experiment for the stream capturing is performed the volume of the file data becomes heavy Be sure to keep enough blank space in the hard disk on the control PC before performing the time lapse experiment For the stream capturing the number of frames varies among the stream blocks 32 Chapter 3 New Time lapse Setting Screen 3 1 5 Time lapse Experiment Scheme Cell Name etc Tab Item Sample name Cell name User name Dimension Load button Save button Sample name Sample Cell name User name Dimension First quadrant test test test Second quadrant test Third quadrant tests Fourth quadrant testa tests test Figure 3 1 18 Cell name etc tab Table 3 1 9 Functions on the Cell name etc tab Function The Sa
112. text Menu on the Title Bar Right click on the title bar of this software window displays the context menu Time lapse images Acquired BioStation IM Version 2 20 Build 140 Move X Close Alt F4 Figure 1 3 1 Context menu on the title bar Table 1 3 1 Functions of context menu Item Function About Bio Station The About BioStation IM dialog box is displayed IM The dialog box showing the version information of this software appears Device information The Device information dialog box is displayed The Device information dialog box allows you to check information about the microscope and camera connected to the control PC Error information The Error information dialog box is displayed The item is enabled when an error occurs and the Error information dialog box allows you to check the cause of the error Preferences The Preferences dialog box is displayed The dialog box allows you to configure basic settings appropriate for the environment Hide control panel Only the left side of the software window remains displayed 1 3 1 Checking the Software Versions Click the About Bio Station IM menu to display the About BioStation IM dialog box This dialog box allows you to check the version of the software About BioStation IM Bio Station V Figure 1 3 2 About BioStation IM dialog box Chapter 1 1 3 2 Checking the Device Information Basic Operations
113. the clicked point For the BS IM MC MOT chamber the Well switch tabs are displayed Click the Well switch tab to show its observation points 3 Live The live observation image of the field of view is displayed observation Click in this screen to move the stage until the clicked point locates at the center of this image display screen 4 Image Select to set tone adjustment saturation check intensity screening and pseudo color adjustment display buttons Operation of each function is the same as that for the Live observation screen 5 Start Select to start time lapse experiment time lapse Time lapse start confirmation dialog box Confirmation window appears button If the Pause button Time lapse images in process screen is used to suspend the time lapse experiment that is running the Start time lapse button changes from Start time lapse to Restart time lapse for restart 6 Time lapse Points tab Select to display the observation conditions filter name exposure time experiment and magnification for each observation point scheme Click an observation point to display the Point information dialog box In the dialog box checking changing and deleting the condition are available Time tab Select to display the capturing interval time total observation time and number of rounds for time lapse experiment Click time lapse experiment time display to display the Timelapse dialog box In the dialog box changi
114. the magnification is Full because the entire tiled image is displayed For the BS IM MC MOT chamber the Well switch tabs are displayed on the right side of the display Click the Well switch tab to see the observation points registered to each well Y Well switch tabs zB845um 344140 Figure 3 2 3 Observation point verification display for the BS IM MC MOT chamber magnification Full 41 Chapter 3 New Time lapse Setting Screen 3 2 2 Browsing or Clearing the Tiled Image The observation area is browsed as a tiled image Clear browse area Browse Browse all Figure 3 2 4 Tiled image display Table 3 2 3 Functions for setting or clearing a browse area on the tiled image Item Function Browse clear Clear browse area Select to clear browsing the tiled image buttons T Select to browse the selected area with the mouse pointer as L Browse 2 tiled image See the figure at lower left Select to browse the entire image as a tiled image See the Browseall figure at lower right Blue frame Registered observation point The number of a registered observation point appears within the frame Yellow frame Current pointer which specifies an observation point Scroll bar Use this scroll bar to scroll the display when the tiled image is enlarged Figure 3 2 5 Tiling images left Browse right Browse all 42 Chapter 3 New Time lapse Setting Screen Browsing
115. the saturation check condition clicked this button again This function can be used only for the fluorescent microscopy Intensity Select to display the Intensity Screening dialog box for setting intensity screening screening button This function can be used only for the fluorescent microscopy Pseudo color button Select to assign a pseudo color for fluorescence images For overlaid images this button is available only when a fluorescence image is used When this button is clicked the saturation check button and the intensity screening button are disabled On the other hand when the saturation check button or the intensity screening button is clicked the pseudo color button is disabled Enlarge reduce zi Enlarge Select to enlarge the image buttons button The enlarge button changes to the reduce button after the Reduce image is enlarged Click the reduce button to reduce the button image to the original size Overlapping mE ____ _ Select to display selected filters overlapped image or buttons mh f display Ratio image Ratio setting Select to set Ratio image button Lu The Ratio setting dialog box appears 53 Chapter 4 Time lapse Images in Process Screen Changing filter name and display color Right click the F11 or FI2 filter button to open the sub menu Filter settings Select the sub menu to display the Filter settings dialog box Filter settings Filter Mame FL1
116. ttings can also be performed on the Simple mode Save button c Select to save observation condition setting in a setting file or in a file for setting on the Simple mode 19 Chapter 2 Live Observation Screen 2 2 1 Saving Observation Conditions Click the Save button 5 in the observation condition setting area to display the Save setting dialog box And then save observation condition setting in a setting file or the Bright Middle or Dark setting file on the Simple mode Setting file compatibility Files created by the software of Ver 2 1 or later cannot be read into the software of Ver 1 Save settines Objective 20x Filter Ph DIA 150 Exposure time 5ms Gain 1 00 Resolution 640 x 480 Binning Save into file Save as Simple Setting for Figure 2 2 2 Save settings dialog box Save into file Table 2 2 2 Functions for saving observation conditions Save into file Item Function Save into file Select to save the observation condition as a setting file Save button Select to display the Windows Save As dialog box Input a file name and save the observation condition as a setting file Close button Select to close the dialog box without saving the observation condition 7 Save into file Save as Simple Setting for C Middle Sample C Dark Sample Save Close Figure 2 2 3 Save settings dialog box Save as Simple Setting for Table 2 2 3 Functions for saving observati
117. tware of the version by which it was originally created 5 1 Channels Display 1 BioStation IM Version 2 20 Build 140 test New es Temp chamber 370 870 Water 380 m time lapse ud m E 20 Stable uw 414 311 207x156um 104x78um Focus X 1 1 3000 33 3000 33 um BLE E File name test Sample name Cell name Qu 2 j Mutich Ph Fn Fe 2 PE 1 Ph Sms FIT 300ms FI2 300ms 20x 2 Ph Sms 300ms FI2 300ms 20x 3 Ph Sms 300ms 2 300ms 20x 4 Ph Sms 300ms 2 300ms 20x Time Range Fit v Time Oh 05m 34s Intensity Nov 26 0500 PM Nov 26 0505 3 20 R 529r Number of rounds 4 1919 1 A 1780 Temperature log vw Qum 4 1641 Chamber 370 370 1501 Water 380 380 Oh 00m 40s Oh 01m 20s Oh 02m 00s Oh 02m 39s Oh 03m 19s Oh 03m 59s Outside 26 0 260 deg P Round 4 E p D g 0 00 SS iV A LL e R dx B T o 6 Figure 5 1 1 Time lapse images Acquired screen Channels display Table 5 1 1 Functions on the Time lapse images Acquired screen Channels display 1 2 Item Function 1 File load button Select to load the saved time lapse result file 2 Observation
118. uct apart moet worden ingezameld Het volgende is alleen van toepassing op gebruikers in Europa Dit product dient gescheiden ingezameld te worden op een daartoe bestemd inzamelpunt Niet wegwerpen bij het normale huisvuil Neem voor meer informatie contact op met het verkooppunt of met de lokale instantie die verantwoordelijk is voor het verwerken van afval Pt S mbolo para recolha de residuos em separado utilizado nos paises Europeus Este s mbolo indica que este produto para ser recolhido separada mente Esta norma aplica se s para os utilizadores nos pa ses Europeus Este produto est designado para recolha de res duos em separado num recipiente apropriado N o deitar no caixote do lixo dom stico e Para mais informa es contactar o revendedor ou as autoridades locais respons veis pela gest o dos res duos I Simbolo per la raccolta differenziata applicabile nei paesi europei Questo simbolo indica che il prodotto va smaltito separatamente La normativa che segue si applica soltanto agli utenti dei paesi europei prodotto designato per lo smaltimento separato negli appositi punti di raccolta Non gettare insieme ai rifiuti domestici Per maggiori informazioni consultare il rivenditore o gli enti locali incaricati della gestione dei rifiuti This symbol is provided for use in the People s Republic of China for environmental protection in the fields of electronic information products 0 2718
119. unction Sample name Input names arbitrary letter strings for the Sample name Cell name and User Cell name name User name Apply button Select to register the input name Close button Select to close the dialog box without registering the input name Inputting name of film bottom 4 quadrant dish Click Dimension field on the Cell name etc tab to displays the Dimension dialog box Dimension First quadrant bes Second quadrant test Third quadrant Fourth quadrant eb Apply Close Figure 3 1 20 Dimension dialog box Table 3 1 11 Functions on the Dimension dialog box Item Function First quadrant Second quadrant Input names arbitrary letter strings for the respective compartments of the film Third quadrant bottom 4 quadrant dish Fourth quadrant Apply button Select to register the input name Close button Select to close the dialog box without registering the input name 34 Chapter 3 New Time lapse Setting Screen 3 1 6 Time lapse Experiment Scheme Zstack Tab Use this function to set up multiple observation points in the Z direction at a same X Y coordinate When the Z stack function is used the setting information is saved and will be restored at the next time When the stream setting is configured on the Time tab the Zstack tab is not displayed Register points with atack Step 050 zi um Donn 2 steps Up 12 From 1 00 um Ta 1 00 Figure 3 1 21 Zstack tab Table 3 1 12
120. user mark to move the playback slider to the user mark position Mark Comment Input Apply Close Figure 4 1 28 Mark Comment Input dialog box This mark is displayed at the point set with the Edit cell stimulus mark setting button Delete Right click the cell stimulus mark to display the submenu Figure 4 1 29 submenu Select Delete from the submenu to delete comments Select Edit to display the Mark Comment Input dialog box with which you can input confirm or change comments Click the Apply button in the Mark Comment Input dialog box to apply the edited comments mark to move the playback Left click the cell stimulus slider to the cell stimulus mark position Mark Comment Input Apply Close Figure 4 1 30 Mark Comment Input dialog box 60 Chapter 4 Time lapse Images in Process Screen 4 1 10 Time lapse Experiment Scheme Points Tab M 1 Fh1 180s FIT 1 48 FI2 1 42 20 BE 2 Ph 1 1805 FM 1 4 FI2 1 48 20 Figure 4 1 31 Points tab Table 4 1 14 Functions on the Points tab Item Function Observation point Select to display the observation conditions filter name exposure time and magnification for each observation point When time lapse experiment is performed only the point selected with a check mark is observed Click an observation point to display the Point information dialog box In the dialog box checking changing and deleting detailed condition are available Sto

Time Lapse Imaging System BioStation IMQ CELL-S2 / CELL-S2

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