User`s Manual Multi Point Time Lapse
Contents
1. Frame 0 Time 0 0 0 Remaining Time T T y Fig 2 2 Display when protocol processor and this software started 2 2 Page Multipoint Time Lapse Observation Connecting This Software and Protocol Processor 2 2 Connecting This Software and Protocol Processor 1 Select With Protocol Processor from Option menu on XY Stage Control window Options Preferences Ctrl F Fig 2 3 Sub menu The TIEMPO group box and Stage Coordinate Information Registration sheet displays the PP Track No and File Name fields E Untitled XYStage SBE File Edit View Options Control Current Pos um x 7570 Y 68180 gp TIEMPO group box Saini rocessin I Step Size um ed by the j ei m0 eyja protocol processor For statement etc here Set to Protocol Processor check box When the lt Add Pos gt button is clicked the creation of the protocol for the added coordinate position is enabled 10000 0 1363 0 2 R0015R001 7272 0 606 0 og 3 ROO2Z RO02 PP Track No field 6818 0 4 ROO3 R003 Shows the track number of the protocol processor corresponding to the current track The protocol processor s track is enabled when this is checked File Name field Specify the image file name to be saved after executing the protocol Fig 2 4 XY Stage Control Window for TIEMPO Co2. 2 9 Saving the Protocol Save the protocol used in the multipoint time lapse observation in the CSV file format For details refer to 2 4 3 Saving the Protocol in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual Page 11 Multipoint Time Lapse Observation Saving the Stage Coordinate Information 2 10 Saving the Stage Coordinate Information Save the stage coordinate information used in the multipoint time lapse observation in the CSV file format In the XY Stage Control window select the File menu Save As name the information file and save it 2 11 Exiting from This Software 2 12 Exiting from the FLUOVIEW software 2 12 Page In the XY Stage Control window select the File menu Exit Exit from the FLUOVIEW software For details refer to 1 2 16 Exiting from the Software in Volume OPERATION INSTRUCTIONS of the FV300 User s Manual or 1 2 11 Exiting from the Software in Volume OPERATION INSTRUCTIONS of the FV500 User s Manual System Requirements System Requirements Specifications Computer system Computer Same as the FLUOVIEW FV300 500 systems Monitor Image input block Image memory Input memo XY stage MPT AS01 FV MPT AS02 FV BIOS series SIGMA KOKI Co Ltd ProScan Prior Scientific Instruments Ltd ProScan is registered trademarks of Prior Scientific Instruments Ltd Page 3 1 OLYMPU
3. Control window 2 Select Open PAPP Track Editor from Edit menu Track window of track to protocol processor that meets with track selected would be displayed For details of Track window see FV300 500 Users manual 2 4 2 4 Input supporting function Copy Ctrl C Paste Ctrl P Delete Ctrl D Register the selected comment as base comment Ctrl B Adjust the shift of Z axis Ctrl 2 Re tegister to PAPP Ctrl R Open PAPP Track Editor Ctrl E _ TIP o Click mouse right side over Stage Coordinates Information Registration sheet and it is possible to select Open PAPP Track Editor or display Track window eee a K TIP To reflect all settings of FLUOVIEW software to the tracks in XY Stage Control window select Re register to PAPP from Edit menu 2 5 Acquiring Images Execute the created protocol to acquire the multipoint time lapse images gt 1 Click the lt Start gt button in the PAPP window Start gt butt Re cabins For details on the protocol execution refer to section 7 4 Executing the Protocol in the FV TIEMPO User s Manual e Page 2 9 Multipoint Time Lapse Observation Z range Offset 2 6 Z range Offset In case that Z position is displaced due to deflection of microscope or ambient temperature change set Z Pos Current Pos once again with use of XY Stage Control window 1 Select registered number of track stage coordinates informatio
4. Enter number of rows in Row text box and number of columns in Column text box and then select lt Set gt button E Define Region by Size x Row 20 Column 20 Fig 1 9 Define Region by Size Dialog box e Page 1 11 Software Description Registering the Stage Coordinate Information 3 The stage coordinate information of each indexed area around the current position is registered automatically in order of the displayed area numbers Ristcenp x Edit View Normal View g Index size Current position 7 The index size can be changed by the user Se ai For details see section 1 7 3 Changing the Index Size If you specify two points start point and end point that are diagonally positioned each other inside region rectangular region to acquire coordinate information the region specified will be equally divided in index size size of view programmed You can therefore acquire adjacent images by moving the stage in every index divided 1 12 Page Software Description Registering the Stage Coordinate Information 1 Select Define Region by Bounds from Edit menu of Stage Map window and the following dialog will appear Define Region by Bounds lt Start Pos gt Button Set the 1st point X and Y coordinates x Pos um Y Pos um Start Pos 4545 0 4545 0 lt End Pos gt Button Set 2nd point X and Y y End Pos 4692 0 5298
5. column Registration Example and enter the name Input data to text box Registration example of File Name Directory name File name No 1 Track No 2 Track ecce No n Track R R R001 R001 R002 R002 ROOn ROOn R R RO1 RO0001 R02 R00002 gt ROn R0000n OLY R OLY RO01 OLY ROO2 OLY ROOn blank OLY OLY OLY svete OLY blank blank Table 1 1 File Name Registration Example Page Multipoint Time Lapse Observation 2 Multipoint Time Lapse Observation Multipoint time lapse observation using a protocol processor is possible when this software is used together with the FV300 or FV500 system and the FV TIEMPO Time Course Software The observation procedure is as shown below For the operations in each step refer to the section or manual mentioned inside parentheses Turn power on and start the FLUOVIEW software Sections 1 2 1 Turning Power ON and 1 2 2 Starting the Software in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual Start this software and protocol processor Section 2 1 Starting This Software and Protocol Processor Connect this software and protocol processor Section 2 2 Connecting This Software and Protocol Processor Start repeated scanning and determine the XY stage coordinate information Section 2 3 Setting the Stage Coordinate Information Edit the pr
6. INSTRUCTIONS of the FV300 User s Manual or section 2 2 1 4 9 Adjusting the image brightness in Volume OPERATION INSTRUCTIONS of the FV500 User s Manual 4 Move the stage to an approximate area near the observation target position In the Stage Map window panel double click on the approximate position you want to move the stage For details see section 1 2 1 Moving the Stage by Specifying the Approximate Position 5 Using the joystick provided with the XY stage move the stage to the approximate area near the position you want to observe 6 Move the stage to the observation position using the arrow buttons in the Control group box in the XY Stage Control window For details see section 1 2 2 Moving the Stage Step by Step 7 Click the lt STOP SCAN gt button on Acquire panel of FLUOVIEW software to stop repeated scanning 8 If XYZ observation is required set it in the Z Stage sub panel in the Acquire panel of the FLUOVIEW software For details see section 2 2 2 1 1 Setting the Z Direction Scanning Area in Volume OPERATION INSTRUCTIONS in the FV300 500 User s Manual e Page 2 5 Multipoint Time Lapse Observation Setting the Stage Coordinate Information Add Pos 9 Click the lt Add Pos gt button in the XY Stage Control window nde Boer Dutton The coordinate information of the current position is added in the Coordinate Information Registration sheet In an i
7. Pos appears here When stage position number registered is selected from drop down list indexes of periphery 5x5 will be displayed enlarged around center of current stage position WERNA Current o Index size Fig 1 6 Stage Map Panel Enlarged The index size can be changed by the user TIPS tan For details see section 1 7 3 Changing the Index Size COE ore ee eth ca ast Ae 2 Double click inside grid of any index at Stage Map so that the stage moves to center of the said index 3 To return to normal display selected the Normal View menu in the Stage Map window e Page 1 7 Software Description Moving the Stage NOTE while the Stage Map panel is displayed enlarged when zoom magnification and object lens magnification are changed on FLUOVIEW software please select the Edit menu Refresh Stage Map will be updated When scanning condition is changed on FLUOVIEW software and to fan out the index size changed it is necessary to update Stage Map panel 2 Moving periphery of stage position registered 1 Select Registration Pos from Large View menu of Stage Map window so that the following dialog box will appear ye Set Registration Pos Registration No fi Set 1 25 Fig 1 7 Set Registration Pos dialog box 2 Enter stage position number registered in Registration No text box and select lt Set gt button In
8. Software and Protocol Processor 1 Select lt Protocol Processor gt button from Time Series sub panel of Acquire panel on FLUOVIEW software Protocol processor boots up 1 il i 1 10 000 0 sec lt Protocol Processor gt button Total Time Click to start this software 1 8 sec and protocol processor Protocol Processor Fig 2 1 Time Series Sub panel in the Acquire Panel of the FLUOVIEW Software 2 This software starts up For further details see 1 1 Starting the software This software Untitled XYStage IIT Ea Stoae Mop Acquire File vO Tile Proce Live SAMPLE XYZT X 3D DFLUOVIEWY Edt View Large View Focus is XY Repeat Once Focus x00 wie ma oe Ne PMT Gain offset ua Step Sit ah en Yp a me on Fa ale nev T fE par Taw 840 0 7 x OT Oxz Cxvzt size 256 by 256 E objective PLAPO AIK UseCA2 PAPP software Image File Name D FLUOVIEW Images 256 by256 10 0 0 0 Noma 0 0 256 256 Fat 00 10 me Image Size Zoom Paa Ansle Sub Mode Sean aser X Pos Pos X Size Y Size Speed M_Ari M_Ar2 m
9. TIEMPO Path Repeat k XY Stage Control Window N PAPP Window 5 4 Start b For Normal Normal K7 2 wy klarmal in 6 PAPP Window _ Image File Name C FLUOVIEW images s Path 1 Interval text box drop down list It works as in the same case as PAPP window 4 Unit of interval time is to be selected from drop down list and interval time to execute protocol by returning from Next track to For track should be put in text box for repeated process of protocol processor 2 Repeat text box It works as in the same case as PAPP window 5 Designate cycles of repeated process for protocol processor 3 lt Path gt button It works as in the same case as PAPP window 6 Designate destination to save image acquired For further details of PAPP window operation see FV300 500 User s Manual 2 4 2 Editing protocol e Page 2 7 Multipoint Time Lapse Observation Editing the Protocol File 2 4 2 Setting information that meets with track selected to FLUOVIEW software It is possible to set the setting value of each track for protocol processor that meets with each track in Stage Coordinates Information Registration on XY Stage Control window to FLUOVIEW software 1 Select Set PP Track Info to FV with position movement from Option menu and make it to the state that a check is entered Options Preferences Ctrl F v with Protoco
10. as plural numbers of coordinate information automatically For further details refer to 1 3 2 Auto Registration of Stage Coordinate Information Specifying Region Refresh Refreshes the enlarged view by obtaining the current image size and index size This item is available when the image is being enlarged View Registration No It indicates number of registration for stage coordinates registered inside index in Stage Map panel Registration FileName It indicates file name inside index in Stage Map panel Large View Current Pos Enlarges the peripheral areas around current stage position in the Stage Map panel For details see section 1 2 5 Moving the Stage to an Indexed Area Registration Pos Enlarges the peripheral areas around stage being registered in Stage Map panel For further details refer to 1 2 5 Moving the Stage to an Indexed Area Normal View Returns the image in the Stage Map panel to the normal size 1 16 Page Software Description Stage Coordinate Information Markings Used in the Stage Map Panel 1 5 Stage Coordinate Information Markings Used in the Stage Map Panel The Stage Map panel uses the following markings to indicate the registered coordinate positions current position etc E Stage Map x Edit View Large View Marking Name Description Origin A red handle indicates the registered origin Current position A
11. be adjusted by changing the index size 1 In the Preference dialog box select Index Size to display the sub panel Based on Scan Size Check box Preference x general Manipulation In case that calculation of index size is required based on scanning size as reference check the check box Ya um X Index Y Index Text box Yindex eum Shows the index sizes in X and Y axis values K ss Shows the percentage of the index size The setting can be changed by direct entry of the value Fig 1 16 Index Size Sub panel Index Size Software IV Based on Scan Size Adjustment slider Page hg x Software Description Changing the Settings In case that calculation of index size is required based on scanning size as reference check the check box of Based on Scan Size When check box of Based on Scan Size is checked you can program the percentage of index size in left side text box of adjuster slider using adjuster slider Depending upon the value set the index size will be calculated and displayed in text boxes X Index and Y Index respectively Actual field White parts are the margins overlapped with the images of adjacent areas Y axis index size Field if 90 index size X axis index size 1000um i In case that the check box of Based on Scan Size is not checked you can enter the value directly in text boxes X Index and Y Inde
12. change the stage control related settings including the stage movement speed stage movement acceleration XY stage connection port etc The basic setting procedure is as follows Select Preferences from Option menu on XY Stage Control window and display Preference dialog Preference x General Manipulation N Index Size j Software Speed r Mechanical Origin Fast 12 0 mm s Go To Mechanical origin Acceleration Connect Using 50 0 mm sec 2 COM 1 7 Connect Fig 1 14 Preference Dialog Box 1 7 1 Setting the Stage Control Parameters Set the stage movement speed stage movement acceleration stage returning to the mechanical origin and connection port as described below 1 Setting the stage movement speed and stage movement acceleration When a quick stage move is required set the value of moving speed and acceleration larger and on the other hand if you wish to minimize the stage move in case of wafer dip observation in a Petri dish it is effective that you set the value of moving speed and acceleration smaller Select Preferences from Option menu on XY Stage Control window and display Preference dialog Preference General Manipulation Speed scale Slow Index Size Software m Mechanical Origin Go To Mechanical origin Select the stage movement speed 12 0 mm s Acceleration Acceleration text box gt 50 0 mm se
13. other observations such as XZ Xt XYZ XYt and XYZt In this manual there are some modified panels and dialog boxes partly changed in window size and gray characters are also modified for enabling users to read them easily e Page 3 NOTES ON THE FLUOVIEW MANUALS NOTES ON THE FLUOVIEW MANUALS This instruction manual is the instruction manual for Multi Point Time NOTE Lapse software to be used on FV300 FV500 or FV TIEMPO Expressing Refer to FV300 500 TIEMPO User s Manual in this manual means that the user is requested to read the instruction manual for the FV300 FV500 or TIEMPO the user possesses NOTE Although the descriptions in this manual are common for the FV300 and FV500 the figures always refer to the time course software for the FV500 BEFORE USE Refer to Volume SAFTY GUIDE in the FV300 500 User s Manual Be sure to read this volume before use SPECIFICATIONS Refer to Volume SAFETY GUIDE in the FV300 500 User s Manual SYSTEM OUTLINE Refer to Volume SYSTEM OUTLINE amp SPECIFICATION in the FV300 500 User s Manual MAINTENANCE Refer to Volume MAINTENANCE in the FV300 500 User s Manual TROUBLE Q amp A Refer to Volume TROUBLE Q amp A in the FV300 500 User s Manual 4 Page Contents Doo 1 Software Description 1 1 1 1 Starting the Software ccccccceecseeeeeeceeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeseeeeeees 1 1 1 2 Movin
14. 0 coordinates A Clear lt Clear gt Button Determine the end point of a eqistration domain Procedures are It returns 1st and 2nd point to explained bere original positions Fig 1 11 Define Region by Bounds Dialog box 2 Move stage to start point of rectangular region and select lt Start Pos gt button 3 Move stage to end point of rectangular region and select lt End Pos gt button Stage coordinate information will be automatically registered in sequence of number inside the display Regarding method of stage move refer to 1 2 Moving the Stage Edit View Large View Fig 1 12 Stage Map Panel with Tiled Indexed Areas Page 1 13 Software Description Menu List 1 4 Menu List Clicking a menu name in the XY Stage Control window or Stage Map window displays the control menu which contains commands for controlling the window 1 4 1 XY Stage Control Window Menu File Edit Sub menu Function New Creates a new stage coordinate information registration sheet Open Loads a CSV file to edit the stage coordinate information Save Saves the stage coordinate information by overwriting previous data Save As Saves the stage coordinate information being edited in a different file name and or location Exit Exits from the application Copy Copies the selected string cell or track into the clipboard Paste Inserts the data in the clipboard in the cursor pos
15. OLYMPUS User s Manual FLUOVIEW Multi Point Time Lapse Multi Point Time Lapse Software Ver 4 3a Thank you for purchasing a XY stage control software To ensure the safety obtain optimum performance and familiarize yourself fully with the use of this product we recommend that you study this manual thoroughly before operate it This manual describes Multi Point Time Lapse software only to be used on FV300 or FV500 Together with this manual please also read FV300 500 TIEMPO User s Manual so that you can understand the overall operation method Keep this manual in an easily accessible place near the work desk for future reference AX6636 CAUTION lt is prohibited to reproduce copy or duplicate a part or the whole of this software and manual The information described in this manual may be subject to change without notice Registered Trademarks Microsoft Microsoft Windows Excel for Windows are registered trademarks of Microsoft Corporation Other brand names and product names are trademarks or registered trademarks of their respective owners e Page NOTATION IN THIS MANUAL NOTATION IN THIS MANUAL This manual has been compiled based on the following notation rule lt Notation of Caution Notes and Tips Notation Description A Caution for preventing injuries of the user or damage to the product including the surroundings NOTE Note for the user TP Hint or one point advice which can be a re
16. S OLYMPUS CORPORATION 2 43 2 Hatagaya Shibuya ku Tokyo Japan OLYMPUS OPTICAL CO EUROPA GMBH Postfach 10 49 08 20034 Hamburg Germany OLYMPUS AMERICA INC 2 Corporate Center Drive Melville NY 11747 3157 U S A OLYMPUS SINGAPORE PTE LTD 491B River Valley Road 12 01 04 Valley Point Office Tower Singapore 248373 OLYMPUS OPTICAL CoO U K LTD 2 8 Honduras Street London E OTX United Kingdom OLYMPUS AUSTRALIA P PTY LTD 104 Ferntree Gully Road Oakleigh Victoria 3166 Australia This publication is printed on recycled paper Printed in Japan 2003 10
17. c 2 Enter the stage movement Connect Using COM 1 vj Connect acceleration value Enter a value in the unit of 1 0 mm sec Page 1 19 Software Description Changing the Settings 2 Returning the stage to the mechanical origin The mechanical origin is the origin point specific to the stage You can set the software origin in the desired position The stage is controlled with reference to an origin but using the mechanism origin as the reference position makes the stage control difficult The stage control can be made easy by setting the software origin in the position you want to use as the reference position for observation Returning the stage to the mechanical origin is executed automatically at the start of this software With certain stages very small errors are accumulated after repeated movements This operation returns the stage to the mechanical origin in order to nullify the accumulated errors After the stage has been returned to the mechanical origin it can be moved from there by the software control Preference Eg General Manipulation Index Size Software r Mechanical Origin lt Go To Mechanical origin gt button Click to return the stage to the mechanical origin Go To Mechanical origin 12 0 mm s Acceleration Connect Using 50 0 mm sec 2 COM 1 7 Connect 3 Setting the connection port This setting is required when changing the con
18. dexes of periphery 5x5 will be displayed enlarged around center of the registered stage position of which number was entered fj Stage Map Edit View Normal View Index size Stage coordinates displayed in the center represents stage number being registered When Current Pos is selected the peripheral indexes 5x5 will be displayed enlarged around center of current stage position Stage position at registered number Fig 1 8 Enlarged Stage Map Panel 1 8 Page Software Description Moving the Stage ercte i TIP a Index size can be changed For further details see section 1 7 3 Changing E ee E a aan ee ee CO TO 3 If you double click any grid of indexes on Stage Map panel the stage will move to center of the said index 4 To return to standard display select Normal View of Stage Map window NOTE While Stage Map panel is displayed enlarged and zoom magnification and objective lens magnification are changed on FLUOVIEW software please select Refresh from Edit menu Stage Map will be updated When scanning condition is changed on FLUOVIEW software and to fan out the index size changed it is necessary to update Stage Map panel One Point Select Registration No from View menu of Stage Map window check mark should appear at the item and the number that stage coordinates are registered will appear Select Registration FileName from View
19. ference for the user lt gt Notation of Menus Command Buttons and Dialog boxes Notation Description Acquire panel The name of a panel dialog box list box or check box is enclosed inside square brackets lt OK gt button The name of a button in a panel or dialog box is enclosed inside angle brackets lt gt lt gt Notation of Mouse Operations Notation Description clicking Action of pressing then immediately releasing the mouse button double clicking Action of clicking the mouse button successively twice dragging Action of moving the mouse while holding the mouse button and releasing the mouse button at the desired position Note In this manual click double click and drag should use left button of the mouse unless otherwise specified Page NOTATION IN THIS MANUAL lt Notation of key operations Notation Description Enter The name of a keyboard key is enclosed inside Alt F1 The positive sign expresses the combination of more than one key operation For example Alt F1 refers to pressing the Alt key while holding the F1 key down Generic names given to the gt N M keys Direction Keys lt Notation of system specific terms Notation Description XY observation Refers to observation with XY scanning other observations The same principle also applies to
20. ftware cccceeeceeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeneeeeees 2 12 2 12 Exiting from the FLUOVIEW softwar e cccccccceeeseeeeeeeeeeees 2 12 3 System Requirements 3 1 Software Description Starting the Software 1 Software Description This chapter describes the functions and setup methods of the software 1 1 Starting the Software 1 Click the lt Start gt button on the bottom right of the Windows screen 2 In case of Windows NT select Programs Windows NT Explorer command from Start menu displayed In case of Windows 2000 select Accessories Windows Explorer command Windows NT or Windows2000 Explorer as shown below will be displayed Y Exploring C FLUOVIEW OO x File Edit View Tools Help z aoe l Le E Fluoview gt aa Hae a e SISA All Folders Contents of C FLUOVIEW My Computer Draw dll Application Extension 9 3 Floppy 4 Driver dll Application Extension CC Help dll Application Extension L Dikar lo dll Application Extension 4 Fluoview Ix ll Application Extension H Program Files Lsm dll Application Extension 5 G aa MicroscopeCore dll Application Extension D Su dll Application Extension E F T askCore dil Application Extension 32 Control Panel TestLog txt Text Document 23 Printers a NDContral ocx OCX File E Eol Network Neighborhood a NewLED ocx OCK File Gy Recycle Bin a OF xmiMonitor ocx OCX File My Briefcase gt
21. g the Stage ssasnsnnnnsnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnmnnn nnne n nnne 1 3 1 2 1 Specifying approximate position ON stage cccceceeeeeeeeeeeeeeeeetesteneeeees 1 3 1 2 2 Moving the Stage Step by Step ccc ccceceeccceeeeeeceeeeeeeeseeneneeeeeeeeeeseenseneeees 1 4 1 2 3 Moving the Stage to the Specified Coordinate Position eeeeee 1 5 1 2 4 Moving the Stage to a Preset Position cee ceeee eee eeee eee eeee teen eeeee teeta 1 6 1 2 5 Moving the Stage to an Indexed Area ecceeee cece eee e eee eeeeeeeeeeeneeeeeneeeeeeeeas 1 7 1 3 Registering the Stage Coordinate Information ceeees 1 10 1 3 1 Auto Registration of Stage Coordinate Information Specifying Index Numbers1 11 1 3 2 Auto Registration of Stage Coordinate Information Specifying Region 1 12 aA Men U SE ar ta aae aap aaa a r a a aan aap e ar i eitan 1 14 1 4 1 XY Stage Control WiNndOW ss sssesssessseesirrsrrrrrrrtrrrtrrrrirrtrrrtrrrtrrrrereenrense 1 14 1 4 2 Stage Map WINdOW oeer eei aa a eaaa eee aar Aa Aa E EE Aa Eh 1 16 1 5 Stage Coordinate Information Markings Used in the Stage Map PAIS E A E A T 1 17 1 6 Enlarging the Stage Map Panel ccccccccceeeeeeeeeeeeeeeeeeeseeeeees 1 18 1 7 Changing the Setting sis siicc soseccccccecte sotes ceded eee ees 1 19 1 7 1 Setting the Stage Control ParameterS cccsccceeceeeeeeeeeeeeeeeteetetenneaees 1 19 1 7 2 Al
22. green handle indicates the current position Registered point White dots indicate the positions registered in the Stage Coordinate Information Registration sheet Registered point When a registered number in the Stage Coordinate Information confirmation Registration sheet is selected a white handle indicates the position corresponding to the registered number e Page 1 17 Software Description Enlarging the Stage Map Panel 1 6 Enlarging the Stage Map Panel When you select Current Pos from Large View menu of Stage Map window the peripheral indexes 5x5 are displayed enlarged around center of current stage position In addition when you select Registration Pos from Large View menu and specify number of stage being registered on dialog box that appears the peripheral indexes 5x5 are displayed enlarged at center of stage being registered For further details refer to 1 2 5 Moving the Stage to an Indexed Area fj Stage Map x Edit View Normal View Current Pos Index size Current position The index size can be changed by the user n For details see section 1 7 3 Changing the Index Size To return from the enlarged display to the normal display select the Normal View menu in the Stage Map panel 1 18 Page Software Description Changing the Settings 1 7 Changing the Settings This section describes how to
23. ignment of moving direction 0 cece eee eee tee teee teen teee teen teeeeeeeee 1 21 1 7 3 Changing the Index Size cececcccceeceee cece eeeeeeeeeceeeeeeeeeeeeseeeneaeeeeeeeetseeeas 1 22 1 7 4 Default setting of file name to be registered automatically 0 0006 1 23 2 Multipoint Time Lapse Observation 2 1 2 1 Starting This Software and Protocol Processor ccccseeee 2 2 2 2 Connecting This Software and Protocol Processor 2 3 2 3 Setting the Stage Coordinate Information 2 seeeeee 2 5 2 4 Editing the Protocol File cccccceeeeeceeeeeeeeeeeeeeeseeeeeeeeneeeeeeeeeeeees 2 7 2 4 1 Setting repeated processing to protocol processor c cccceeeeeeeeeeeeeees 2 7 2 4 2 Setting information that meets with track selected to FLUOVIEW software 2 8 2 4 3 Display of input supporting function to track of protocol processot 2 8 2 5 Acquiring IMAageS au o aon ns eee tenet veeenenee dee cte nc ceeteeetene rene soneeneente 2 9 Contents Doo O 2 6 Z range Offset osiin noes Seca dineteeecexcticcednntee oe cent ee ec 2 10 2 7 Editing Images ei oseec es sccs hesearees eonscricss oon eee ter eeve cetacean cer emeiens 2 11 2 8 Saving Mage S niipea aia en eee eee 2 11 2 9 Saving the Protocol sssssssssusnnnunnnnnnnnnnnnnnnnnnnnnunnnnnnnnnnnnnnnnn nnmnnn 2 11 2 10 Saving the Stage Coordinate Information ccceeeeeeeeeeees 2 12 2 11 Exiting from This So
24. inates of track of SE the track being selected in sheet Add Pos Stage Coordinate Information Registration sheet Stage coordinate information registered will be displayed in X Y and Z axis You can change each setting if you input in each text box directly The Z axis setting is displayed when FLUOVIEW software is started The displayed values are obtained from the settings made with the FLUOVIEW software When No column of track in sheet is double clicked the stage moves to the coordinate of track being selected No 1 2 3 4 5 6 i 8 9 Fig 1 5 Main Screen The stage coordinate information contains the X and Y axis coordinates of the registered position It also includes the Z axis coordinate when the FV300 500 is used 1 6 Page Software Description Moving the Stage 1 2 5 Moving the Stage to an Indexed Area This operation allows the stage to be moved to one of the indexed areas each of which having the specified index size field size This operation is effective when you want to acquire images of adjacent areas on the stage by moving the stage position to the desired areas 1 Moving the periphery of current stage position 1 Select Current Pos from Large View menu of Stage Map window so that peripheral indexes 5x5 will be displayed enlarged around center of current stage position fj Stage Map x Edit View Normal View Current
25. ing of details With Protocol Processor It appears when FLUOVIEW software is linked Connects or disconnects the protocol processor When this is checked the software is connected and the TIEMPO group box and item TIEMPO in the Stage Coordinate Information Registration sheet are displayed Set PP Track Info to FV with position movement It appears when FLUOVIEW software is linked It sets setting information that meets with the track selected to FOLUOVIEW software For further details see 2 4 2 Setting information that meets with track selected to FLUOVIEW software Page 1 15 Software Description Menu List 1 4 2 Stage Map Window Menu Sub menu Function Edit Register Software Origin Registers the current position as the origin Add Current Position Registers the current position in the Stage Coordinate Information Registration sheet Define Region By Size Registers the coordinate positions of the grids indexes around center of the current position in the Stage Map panel divided according to the index size view size automatically as stage coordinate information For further details refer to section 1 3 1 Auto Registration of Stage Coordinate Information Specifying Index Numbers By Bounds Registers positions divided per index size view size specified in the region that is specified as region to acquire coordinate information in Stage Map panel
26. ition Delete Deletes the selected string cell or track Re register to PAPP It appears when FLUOVIEW software is linked Re registers the stage coordinate information in the protocol processor Register the selected comment as base comment When stage coordinate information is newly added to Stage Coordinate Information Registration sheet it programs default comment registered in Comment column Adjust the shift of Z axis It appears when FLUOVIEW software is linked When Z position is displaced due to deflection of microscope or ambient temperature change it programs the shift stroke of Z axis if Z range is adjusted Open PAPP Track Editor It appears when FLUOVIEW software is linked It displays input supporting function to the track of protocol processor For further details see 2 4 3 Display of input supporting function to track of protocol processor 1 14 Page Software Description Menu List Menu Sub menu Function View On Top When this is checked the XY Stage Control window is always displayed at the front Registration When this is checked the Registration group box is displayed Stage Map When this is checked the Stage Map panel is displayed Separate Stage Map When this is checked the Stage Map panel can be moved independently from the XY Stage Control panel Options Preferences Displays the Preference dialog box for use in sett
27. l Processor Ctrlew Set PP Track Info to FY with position movement Ctrl I 2 Double click No column of the track to be set from Stage Coordinates Information Registration on XY Stage Control window The setting information of track to protocol processor will be set to FLUOVIEW software Simultaneously the stage moves to the position where the track is selected NOTE Set PP Track Info to FV with position movement does not appear unless With Protocol Processor of Option menu is checked NOTE If No column is double clicked in the state the Set PP Track Info to FV with position movement is not checked the stage moves to the position where the track is selected but setting to FLUOVIEW software is not executed 2 4 3 Display of input supporting function to track of protocol processor 2 8 Page Track window that is input supporting function to protocol processor can be displayed from XY Stage Control window By checking check box of each group on Track window it is possible to get or set necessary information only from or to FLUOVIEW software The setting information that differs between PAPP window and FLUOVIEW will be highlighted and displayed in orange For details Multipoint Time Lapse Observation Acquiring Images of Track window see FV300 500 User s manual 2 4 2 4 Input supporting function Display method 1 Select track in Stage Coordinates Information Registration on XY Stage
28. ls refer to 1 2 12 7 Setting the Multiple sections to be Observed in Volume OPERATION INSTRUCTIONS of the FV300 User s Manual or 1 2 8 7 Setting the Multiple sections to be Observed in 2 10 Page Multipoint Time Lapse Observation Editing Images Volume OPERATION INSTRUCTIONS of the FV500 User s Manual 6 Upon completion of Z position adjustment select lt Set gt button on Adjust the shift of Z axis dialog box Shift distance of Z position will be registered in Z Pos column of Stage Coordinates Information Registration sheet of XY Stage Software 7 Select lt STOP SCAN gt button on Acquire panel of FLUOVIEW software so that the repeat scanning will be stopped etme Pid TIP Mi In case that the shift distance of Z position is already known you may ars enter the shift value directly in Z Shift text box on Adjust the shift of z _axis dialog box without executing scan and moving the stage to acquire NOTE When you use FV300 500 system that Z motor is not equipped you cannot use Adjust the shift of Z axis function 2 7 Editing Images Edit the acquired XYZ or XYZT images For details refer to 2 6 Image Processing and after in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual 2 8 Saving Images Save the edited images For details refer to 2 3 1 Saving Images and after in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual
29. menu check mark should appear at the item and file name will appear inside the index grid Normal View File Name Registered number 2 R002 R008 Page 1 9 Software Description Registering the Stage Coordinate Information 1 3 Registering the Stage Coordinate Information The stage coordinate information contains the X and Y axis coordinates of the registered position It also includes the Z axis coordinate when the FV300 500 is used 1 Move the stage to the desired position as described in section1 2 Moving the Stage 2 Click the lt Add Pos gt button in the XY Stage Control window The coordinates of the current position are added to the Stage Coordinate Information Registration sheet E Untitled XYStage ME Ei File Edit View Options j Control Current Pos um lt Add Pos gt button xi oof a Click to add the current stage position XYZ coordinates as i i Step Size um the stage coordinate information in the sheet fE ae x 500 Y 50 0 Up down buttons hace istragn Click to move the stage to the f coordinate position of the track Add Pos before or after the cell being registered in the sheet FA o w 0o woon e Om Stage Coordinate Information Registration sheet Stage coordinate information registered will be displayed in X Y and Z axis You can change each setting when you input in each text box directly The Z axis setting is dis
30. n to adjust Z position on Stage Coordinates Information Registration sheet on XY Stage Control window and double click it to move the stage 2 Select Adjust the shift of Z axis from Edit menu of XY Stage Control window so that the following dialog box will appear Adjust the shift of Z axis xi Z Shift 0 00 um Set scanning start position is shifted lt Tip Do mouse right click over Stage Coordinates Informatio 7 Registration sheet on XY Stage Control window and select Adju the shift of Z axis so that the same operation as described above can 3 Select lt XY Repeat gt button on Acquire panel of FLUOVIEW software to execute repeat scanning Acquired image will appear in Live panel For details refer to 1 2 12 6 Repeated Scanning Operation in Volume OPERATION INSTRUCTIONS of the FV300 Users Manual or 1 2 8 6 Repeated Scanning Operation in Volume OPERATION INSTRUCTIONS of the FV500 User s Manual 4 Select Z Stage sub panel on Acquire panel of FLUOVIEW software 5 Watching Live panel adjust Z position using lt Z stage coarse adjustment gt button or lt Z stage fine adjustment gt button on Z Stage sub panel Shift distance of Z position the difference between the value in Current Pos text box and the value in Z Pos of track selected with XY Stage Software will be displayed in Z Shift text box of Adjust the shift of Z axis dialog box For detai
31. nection port or connecting a new stage 1 After the port to which the XY stage has been changed open the drop down list in the Connect Using group box select the new connection port and click the lt Connect gt button General Manipulation Index Size Software p Speed Mechanical Origin Fast Go To Mechanical origin 12 0 mm s Connect Using Acceleration r Connect Using group box 50 0 mm sec 2 ama Select the connection port name from the drop down list and click the lt Connect gt button 1 20 Page Software Description Changing the Settings 1 7 2 Alignment of moving direction Alignment of moving direction is done When the stage is moved in the state that it is not precisely connected it would move to X or Y direction as it has been displaced from the specimen By aligning moving direction the displacement with coordinate can be compensated Alignment should be executed by programming two points on specimen as reference in X direction 1 In the Preference dialog box select Manipulation to display the sub panel lt 1st Pos gt Button Set 1st point on X direction X and Y coordinates Preference eneral Manipulation Index Size Software x um Y um 1stPos 0 0 0 0 lt 2nd Pos gt Button k 2nd Pi Set 2nd point on X Tzara oe ee direction X and Y coordinates Procedures are explained here Clear lt Clear g
32. nnection Mode e Page 2 3 Multipoint Time Lapse Observation Connecting This Software and Protocol Processor 2 4 Page One Point Stop when an error occurs Ctrl F5 v Alarm an Interval remaining time Ctrl F6 Stop an interval remaining timer during pause Ctrl F7 Use R 232 C command in protocol Ctrl F8 COM Control Ctrl F9 v Start up with Y Stage Ctrl F11 Refresh When Changing Mode Ctrl F12 Click Option menu on PAPP window and put a check on Start up with XY Stage and boot up protocol processor so that this software would automatically start up and protocol processor would be connected Multipoint Time Lapse Observation Setting the Stage Coordinate Information 2 3 Setting the Stage Coordinate Information While performing repeated scanning set the stage coordinate information in the Registration group box 1 Set the conditions for repeated scanning in the Acquire panel of the FLUOVIEW software For details refer to section 1 2 Outline of LSM Observation Procedures in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual N Click the lt XY Repeat gt button in the Acquire panel of the FLUOVIEW software lt XY Repeat gt button to execute repeated scanning The acquired image is displayed in the Live panel 3 Adjust the image brightness For details refer to section 2 2 1 3 9 Adjusting the image brightness in lt Focus gt button Volume OPERATION
33. nterlocked operation with this software the protocol is also added in the sheet in the PAPP window 10 Repeat steps 1 to 6 for each of the coordinate positions to be observed This software and protocol processor are set up as follows E Untitled XYStage Miimi Ea E Stage Map x File Edit View Options Edit View Large View Control Current Pos um x 00 0 0 Go a Step Size um SAIL x a elo a TIEMPO Interval 10 0 jsec Pah Repeat 1 Registration gt AddPos p Sette Protocol No Pos _Y Pos ZPos PP Track NO _ File Name 1 636 4 636 4 oM 2 ROOWROO1 2 3182 636 4 ov 3 ROOZ RO02 3 0 0 636 4 ov 4 ROO3 R003 4 318 2 636 4 oo 5 ROOs RO04 5 636 4 635 4 aoM 6 ROOS RO0S 6 636 4 318 2 ov 7 ROOB ROOG 7 318 2 318 2 oo M 8 R0075R007 8 00 3182 ooM 9 ROOB RO08 3 3182 318 2 ov 10 ROOS RO09 10 636 4 318 2 ooMv 11 ROINROIO oy lw id 3 Fig 2 5 XY Stage Control window where the coordinate information is registered File Edt View Run Option Tools w n gt File Comment Image File Name D FLUOVIEW Images Path aE GetIntVaues Init vat 0 2 56by256 1 0 0 0 0 Noma 0 O 256 256 Fast 0 0 1 0 j j Track Frame gt Pan Status Scan Status Scan Laser 4 Registe ring p No Mode Start by Condi
34. openFLUOVIEW dll Application Extension iE PAPP exe Application PHO_Test ini Configuration Settings gt Physiology dll Application Extension ls QAF dll Application a TK 0CK OCX File x ScpSetup exe 1 732KB Application s sidxvb dll 224KB Application Extension gt sysstat dll 116KB Application Extension TeeChart ocx 1 496KB OCX File la test csv 1KB CSV File a TurretControl ocx 107KB OCX File widat exe 52KB Application 5 365KB Application XYStage exe 83 object s 42 1MB Disk free space 2 44GB 3 Double click XYStage exe of Fluoview folder at the drive where FLUOVIEW software is installed e Page 1 1 Software Description Starting the Software 4 Message to confirm stage move to home position and escape of objective lens from stage appears Click lt OK gt button and the stage will move to mechanical home position Detect the mechanical origin check box Remove check when stage is not moved to mechanical origin xY Stage warning The stage will move in a wide range to detect its mechanical origin Escape the objective to the safety position to avoid collison with stage JV Detect the mechanical origine Escape automatically check box JV Escape automatically Remove the check when you do not escape the objective lens from the stage when using the Motorized microscope 5 The main screen of the software shown below appears lt Exit gt button S
35. otocol file Sections 2 4 1 Editing the Protocol and 2 4 5 Loading a Protocol in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual Set the interval and number of protocol repetitions Section 2 4 Editing the Protocol File Acquire Images Using the Protocol Processor XYZ and XYZT Images Section 2 5 Acquiring Images Edit the XYZ and XYZT images Section 2 8 1 Building Extended Focus Image from XYZ Images in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual Save the images Section 2 3 Saving Opening and Shredding Images in Volume OPERATION INSTRUCTIONS of the FV300 500 User s Manual Save the protocol Section 2 4 3 Saving the Protocol in Volume OPERATION INSTRUCTIONS of the FV300 500 Users Manual Save the stage coordinate information Section 2 10 Saving the Stage Coordinate Information Exit from this software Section 2 11 Exiting from This Software Exit from the FLUOVIEW software and turn power off Sections 1 2 16 Exiting from the Software and 1 2 16 Turning Power OFF in Volume OPERATION INSTRUCTIONS of the FV300 User s Manual Sections 1 2 11 Exiting from the Software and 1 2 12 Turning Power OFF in Volume OPERATION INSTRUCTIONS of the FV500 User s Manual e Page 2 1 Multipoint Time Lapse Observation Starting This Software and Protocol Processor 2 1 Starting This
36. played when FLUOVIEW software is started The displayed values are obtained from the settings made with the FLUOVIEW software In Comment column you may enter comment regarding stage coordinates 10 Register other desired observation positions by moving the stage to each of them and adding its stage coordinate information 1 10 Page Software Description Registering the Stage Coordinate Information If you specify reference position as center of region center of rectangular region to acquire coordinate information and number of indexes you can program the region to acquire coordinate information You can acquire adjacent images by moving the stage in every index size size of view programmed inside the region 1 In the panel of the Stage Map window click the mouse on the position you want to set as the reference position A green handle appears 2 Select Define Region by Size from Edit menu of Stage Map window and the following sub menu will appear Select number of indexes column by row to queue View Large View Register Software Origin Ctrl S Add Current Position Ctr 1x3 1 5 Define Region Fag Select the number of indexes to be displayed Sel F4 column x row from the oho ers sub menu 345 FG 5x1 oF 5x3 FB PRT SRES More F12 Select More from sub menu and you can program row number and column number you desire The following dialog box will appear
37. ption Moving the Stage 1 2 2 Moving the Stage Step by Step Move stage per step specified It is useful when you wish to move the position of observation in small movement Arrow buttons Stage moves in arrow direction The movement step is the value entere in the Step Size text box button Click to return the stage to the set origin File Edit View Options 0 0 0 0 Go Registration won ooN e wN eI 4 lt GO gt button Functions as the stop button during stage movement Click if you want to stop movement Step Size text box Set the distance of stage movement step per click of an arrow button Fig 1 3 XY Stage Control Window If you want to register this position as the stage coordinate information refer to section 1 3 Registering the Stage Coordinate Information 1 4 Page Software Description Moving the Stage 1 2 3 Moving the Stage to the Specified Coordinate Position The following operation allows the stage to be moved to the observation position by specifying its coordinates This operation is effective when the coordinates of the observation position is already known and you want to move the stage there directly 1 Enter the coordinates of position to which you wish to move the stage in X axis and Y axis at Control Pos text box and click lt Go gt button E Untitled XYStage SBE Eile Edit View Options Control Pos tex
38. t Button Move to 2st point on new X axis Then click 2st Pos button It returns 1st and 2nd point coordinates or Fig 1 15 Manipulation Sub panel alignment to original positions Move stage to the 1st point in X direction and select lt 1st Pos gt button Move stage to the 2nd point in X direction and select lt 2nd Pos gt button The straight line that passes through two points will be aligned as X axis Regarding method of stage move refer to 1 2 Moving the Stage Ym 1st point When the 1st and 2nd points are specified the software performs the following processing 1 Assumes a right angled triangle having the line connecting the two points as the hypotenuse and computes sin and cos 9 of this triangle e Page 1 21 Software Description Changing the Settings X2 Y2 p Y2 Y1 X1 Y1 X2 X1 2 Performs affine transformation using the obtained trigonometric functions 3 Based on the results obtained above corrects the error in coordinates between the stage and specimen as shown below Overall movement area Xp Yp Software coordinates Aligned stage coordinates 1 7 3 Changing the Index Size 1 22 When connecting the images of adjacent areas after acquiring them certain software is incapable of connecting them properly if the images of adjacent areas do not have some overlapped areas The degree of overlapping between the images of adjacent area can
39. t box Control lt GO gt button Enter the coordinates of the Curent Pos fum When clicked the stage move position in the X and Y oo oo coy moves to the nie text boxes displayed in the Current Pos text box During stage in Step Size um x 500 motion this button changes y 500 to lt Stop gt button Click jie o gt e lt Stop gt button when you r Registration wish to stop gt Add Pos No _x Pos YPos _Z Pos 1 2 3 4 5 6 7 8 9 10 X 4 L Fig 1 4 XY Stage Control Window If you want to register this position as the stage coordinate information refer to section 1 3 Registering the Stage Coordinate Information Page 1 5 Software Description Moving the Stage 1 2 4 Moving the Stage to a Preset Position This operation allows the stage to be moved to one of the coordinate positions set in advance This operation requires stage coordinate information to be registered in advance For details see section1 3 Registering the Stage Coordinate Information 1 Double click column of No of which track coordinate that is required to move is memorized in Stage Coordinate Information Registration sheet on XY Stage Control window E Untitled XYStage Bik File Edit View Options r Control Current Pos um xp oo Yf oo Go Step Size um Up down buttons el x Stage is moved to forward up or IEAI Ap backward down coord
40. tage Map Click to exit from the software window Shows the coordinate lt Close gt button lt Minimize gt button positions including the Click to close the It makes window to minimum current stage position Stage Map and registered points size by turning it to icon window E Untitled XYStage Biim ki ihe File Edit View Options Control XY Stage Control Current Pos um window 0 0 Y 0 0 Go Window for use in registration confirmation and editing of stage Step Size um coordinate information by gt x 50 0 controlling the stage yi 50 0 Registration Pee No x Pos YPos ZPos lt w o woo N e w Nl 3 4 Fig 1 1 Main Window 1 2 Page Software Description Moving the Stage 1 2 Moving the Stage Move stage to the position where the specimen is observed 1 2 1 Specifying approximate position on stage Move stage directly to the position of observation It is useful to move the stage to approximate position before specifying the accurate position Double click the position where you wish to move on panel of Stage Map window ap Large View Edt View Stage Map panel i Double click the position you wish to move inside the panel Fig 1 2 Stage Map Window If you want to register this position as the stage coordinate information refer to section 1 3 Registering the Stage Coordinate Information Page 1 3 Software Descri
41. tion Interval Interval age Size Zoom gt pan Y Pan 209 Sub Mode pos Y pos X Size Y Size Speed M_Ari M_Ar2 M coordinate mi For Normal i 1To1 10 0 a inform ation using M2 XY Normal 1 04 0 256 by 256 1 0 0 0 0 Normal 0 0 256 256 Fast 0 0 1 0 Ot A F3 x Normal 1 0 4 256 by 256 Normal this software results ma xv Normal 1 04 256 by 256 Normal in automatic creation Ws xy Namal 1 ee 256 by 256 Normal ey me xy Nomal 1 0 4 256 by 256 Normal of a For statement M7 x Normal 1 0 4 256 by 256 Normal vs x Normal 1 0 4 256 by 256 Normal vs x Normal 1 oa 256 by 256 a Normal mio xY Normal 1 o4 256 by 256 Normal M1 xY Normal 1 0 4 256 by 256 Normal m12 x Normal 1 0 4 256 by 256 Normal m13 xv Normal 1 0 4 256 by 256 Normal mis xy Normal 1 04 z 256 by 256 Normal a 4 gt Total Frame 25 Time 0 0 10 Remaining Time 7 Fig 2 6 PAPP window where the protocol is set up automatically by registration of coordinate information 2 6 Page Multipoint Time Lapse Observation Editing the Protocol File 2 4 Editing the Protocol File Edit the protocol file to be used in the multipoint time lapse editing For details refer to section 2 4 2 Editing the Protocol in the FV 300 500 User s Manual 2 4 1 Setting repeated processing to protocol processor The repeated processing of the protocol processor can also be set in from the TIEMPO group box in the XY Stage Control window 1
42. x respectively After completing all settings click the lt Close gt button on the top right of the dialog lt Close gt button box to close it 1 7 4 Default setting of file name to be registered automatically Format of file name to be registration of stage position registered automatically in File Name column is set at 1 Select Software panel of Preference dialog box Directory name Check box Text box When you register directory for save automatically check this box and enter directory name in text box File name Check box Text box When you register file name for save automatically check this box and enter file name in text box Q Preference eneral Manipulation E Index Size Default File Name for registration position MV Directory name Ree V File name Re Message Fig 1 17 Software Sub panel Page 1 23 Software Description Changing the Settings 1 24 When you register directory for save automatically check the check box of Directory name and enter directory name in the text box When you assign consecutive number to directory name see the following table 1 1 File Name column Registration Example and enter the name When you register file name for save automatically check the check box of File Name and enter file name in the text box When you assign consecutive number to file name see the following table 1 1 File Name
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