Olympus FLUOVIEW FV300, User Manual

Page 1: ...IEW OPERATION INSTRUCTIONS MAINTENANCE and TROUBLESHOOTING Together with this manual please also read the User s manual FLUOVIEW FV300 and the instruction manual of the microscope in order to understa...

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Page 3: ...ted 2 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 Corpor...

Page 4: ...ECTION MEASURES ACCORDING TO IEC60825 1 1993 A1 1997 A2 2001 LASER PRODUCT RADIATION SAFETY STANDARD I 4 1 II SPECIFICATIONS Another manual Describes the specifications of the FLUOVIEW system 1 SPECIF...

Page 5: ...ist of Functions in the Active Overlays Dialog Box IV I 1 Appendix J Hand Switch and Microscope Frame Function Allocation IV J 1 V MAINTENANCE This manual Describes maintenance of the FLUOVIEW system...

Page 6: ...alog box list box or check box is enclosed inside square brackets OK button Open File button The name of a button in a panel or dialog box is enclosed inside angled brackets Notation of Mouse Operatio...

Page 7: ...eys Generic names given to the o m n and p keys Notation of system specific terms Notation Description XY observation Other observations Refers to observation with XY scanning The same principle also...

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Page 9: ...is volume describes the outline of the FLUOVIEW FV300 system Please read this volume so that you can understand the system before use On This Volume I II II I S SY YS ST TE EM M O OV VE ER RV VI IE EW...

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Page 11: ...on 1 4 1 4 1 System Component Units and Their Functions 1 4 1 4 2 Block Diagram 1 7 1 5 Software Functional Configuration 1 10 1 5 1 Function Panel and Display Panel 1 10 1 5 2 Panel Structure of the...

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Page 13: ...g light detector and outputs the image of the specimen on an image monitor The confocal optics place a confocal aperture at a position which is optically conjugate with the focusing position i e confo...

Page 14: ...focal aperture can be selected from 5 positions so the optimum confocal aperture for each objective can be selected with one touch operation 7 High speed scanning acquires 4 images per second 512 x 51...

Page 15: ...oic mirror Imaging lens Objective Specimen Pupil lens Mirror Barrier filters Laser Light axis adjustment plate Condensing lens Beam expander CH1 PM1 CH2 PM2 Custom dichroic mirror for separating the f...

Page 16: ...laser oscillator Reflected light power supply unit Supply power to the mercury burner light unit Laser Combiner Unit which compounds various laser light Computer Used to control the LSM process image...

Page 17: ...SYSTEM OVERVIEW System Configuration III SYSTEM OVERVIEW III 1 5 Page Photo 1 2 Combination with BX51WI Microscope BX51WI stage fixed upright microscope set up for fluorescence observation...

Page 18: ...Configuration III SYSTEM OVERVIEW III 1 6 Page Photo 1 3 Combination with IX Microscope IX81 inverted microscope set up for fluorescence observation Transmitted light detector optional Unit for acquir...

Page 19: ...alvano X Y cables 2 9 m each 8pin Power control cable 2 9m Analog signal cable 2 9 m 1 8m 5 5m SU control cable 2m Ar LASER POWER SUPPLY UNIT 5 5m HeNe G LASER POWER SUPPLY UNIT 1 8m 1 8m Galvano cont...

Page 20: ...I O cable 2 9 m 50pin Dsub15pin Dsub9pin Galvano X Y cables 2 9 m each 8pin Power control cable 2 9m Analog signal cable 2 9 m 1 8m 5 5m SU control cable 2m Ar LASER POWER SUPPLY UNIT 5 5m HeNe G LAS...

Page 21: ...pin For switching signals output from the left connectors VIS1 and VIS2 are for invisible laser UV is for UV laser IR is for IR laser and Blanking is for switching laser ON OFF UV laser and IR laser a...

Page 22: ...tion to be executed just like when using a system notebook or file folder 1 5 1 Function Panel and Display Panel The FLUOVIEW software is organized by two kinds of panels the function panels and displ...

Page 23: ...age acquisition File I O Tile Process Math Filters Histogram Analyze Single Save load and delete images Change the image display method Process the acquired images Perform mathematical operations betw...

Page 24: ...xecuted by Dragging Dropping This software selects image files and observation methods dye name by means of dragging dropping This allows simple selection based on an intuitive operation of selecting...

Page 25: ...splayed in the Icon field in the Files list box in the File I O panel or during dragging of an image file Use these icons to identify the observation methods used in image acquisition In all observati...

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Page 27: ...g procedures of the FLUOVIEW FV300 system Getting Started FLUOVIEW contains information on the basic operation flow until acquisition of XY images APPLIED OPERATIONS provides detailed operating proced...

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Page 29: ...scope IX81 1 36 1 2 4 4 Combination with Inverted Microscope IX70 1 38 1 2 5 Setting the Dyeing Methods 1 40 1 2 6 Selecting the CONFOCAL APERTURE 1 42 1 2 7 Selecting the DETECTION MODE 1 42 1 2 8 Se...

Page 30: ...on and Jog Sensitivity Adjustment When the BX or IX is used 1 72 2 APPLIED OPERATIONS 2 1 2 1 General Operation Procedure 2 1 2 1 1 Image Acquisition Procedure Section A 2 3 2 1 2 Image Acquisition Pr...

Page 31: ...on of a Line at Desired Angle 2 82 2 2 10 Display the change of image intensity Point Scan 2 85 2 2 11 Image Acquisition on Desired Line XZ XT or XZT Observation 2 89 2 2 12 Image Acquisition in the L...

Page 32: ...otocol using repetition processing 2 180 2 4 9 Pop up Menu 2 188 2 4 10 Restrictions of PAPP setting 2 189 1 Restrictions with Mode and I O card 2 189 2 Restrictions with Mode and Sub Mode 2 191 2 5 C...

Page 33: ...n Image 2 214 2 6 Image Processing 2 215 2 6 1 Filtering 2 215 2 6 1 1 Contour Enhancement 2 216 2 6 1 2 Noise Reduction 2 219 2 6 1 3 Image Sharpening 2 220 2 6 1 4 Correcting DIC Level Irregularitie...

Page 34: ...uilt Image 2 277 2 8 1 2 Turning Built Image into Single Image 2 280 2 8 1 3 Turning Built Image into time series image 2 282 2 8 2 Building line images to be viewed in Z direction 2 282 2 9 Viewing 3...

Page 35: ...12 Changing the Comment Font 2 315 2 13 Image Output at Printer 2 315 2 14 Merger Extraction of Image Channels 2 317 2 14 1 Setting the Range of Multiple Image Slices 2 317 2 14 2 Merging Image Chann...

Page 36: ...O Panel H 1 Appendix I List of Functions in the Active Overlays Dialog Box I 1 Appendix I 1 Coordinate Position Data I 1 Appendix I 1 1 X Coordinate I 1 Appendix I 1 2 Y Coordinate I 2 Appendix I 1 3...

Page 37: ...and Microscope Frame Function Allocation J 1 Appendix J 1 Hand Switch Functions J 1 Appendix J 2 Microscope Frame Functions J 2 Appendix J 2 1 BX J 2 Appendix J 2 2 IX J 3 Appendix J 2 3 Focus Adjust...

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Page 39: ...in use Select the optimum confocal aperture number for the objective being displayed in the control panel displayed by the computer Acquire panel When the acquired image is bright enough and noise is...

Page 40: ...mbers recommended for them Objective N A Confocal Aperture Number PLAPO 40X 0 95 2 PLAPO 60XO 1 4 2 PLAPO 100XO 1 4 3 PLAPO 40XWLSM 0 9 2 PLAPO 60XWLSM 1 0 3 PLAPO 60XOLSM 1 1 3 UPLAPO 10X 0 4 1 UPLAP...

Page 41: ...ic with a shorter wavelength than 570 nm such as FITC x When the slider switch is pushed in half way to the position The custom dichromatic mirror for separating the fluorescence emission is engaged i...

Page 42: ...lens Objective Specime Conde nsing lens Laser Light axis adjustment plate Pupil lens Beam expander CH1 CH2 FL Slider in the pulled out positions Custom dichromatic mirror for separating the fluorescen...

Page 43: ...set pushed in harf way to the position by mistake no images can be observed at all Be sure to set it to the fully pulled out or pushed in position 4 Switching the excitation DM One of the two DMs can...

Page 44: ...n when observing a double labeled specimen such as FITC PI 4 568 488 AT25 Excites the specimen at 488 nm and 568 nm but the excitation output at 488 nm is attenuated to 25 of the normal level Use this...

Page 45: ...Started FLUOVIEW Basic Operations IV OPERATION INSTRUCTIONS IV 1 7 Page 400 500 600 700 wavelength nm CH2 CH1 TRITC 488 FITC Attenuates the excitation intensity to reduce leakage of the FITC emission...

Page 46: ...he microscope refer to the instruction manual of your microscope Combination with BX51 or BX61 1 Light path selector knob 2 Cube turret Cube display window This figure shows the cube display window 3...

Page 47: ...ce observation or transmitted polarized light observation x For laser microscope always disengage the U MDICT3 Engaging the U MDICT3 in the light path degrades the image quality 4 Transmitted light DI...

Page 48: ...erence observation polarizer are also provided x To perform differential interference observation engage the transmitted light DIC slider optional matching the objective in use in the light path For b...

Page 49: ...ination with FVX LT this knob is not attached 1 Light path selector knob BI LSM 2 Universal vertical illuminator mirror cube housing AX URBC 3 Analyzer AX AN optional 4 Transmitted light DIC slider U...

Page 50: ...zing plate for use in differential interference observation and polarized light observation x Set to the pushed in position to engage the AX AN in the light path for direct transmitted light different...

Page 51: ...ry turret for the transmitted light DIC slider and the polarizing plate for differential interference observation polarizer are also provided x To perform differential interference observation engage...

Page 52: ...NSTRUCTIONS IV 1 1 4 Page Combination with IX81 FVF 1 Light path selector button 2 Cube turret Analyzer IX2 MDICT Optionally installed 4 Transmitted light DIC slider U DICTS optional 6 Filters 5 Conde...

Page 53: ...into the light path for direct transmitted light differential interference observation or transmitted polarized light observation x Be sure to disengage the IX2 MDICT analyzer for laser microscopy Eng...

Page 54: ...direct observation and laser differential interference observation x To perform direct differential interference observation or laser differential interference observation engage the polarizing plate...

Page 55: ...elect something and execute its function Double clicking To double click place the mouse pointer on the desired function and press the mouse button successively twice To move something Dragging To dra...

Page 56: ...on a large scale Clicking the top or bottom arrow button allows fine adjustment of the value Dragging the square knob allows the value to vary directly Page tab Click to switch the panel for executin...

Page 57: ...is box displays a control menu which contains the commands for use in controlling the window Group box The group box is organized by grouping functions with the specific meanings and enclosing them in...

Page 58: ...ect the light path for 100 binocular tube and focus on the specimen Section 1 2 3 If no image is displayed If an image is displayed Adjust PMT Voltage Sec 2 2 1 3 9 Change the ND filter with a filter...

Page 59: ...Sec 1 2 10 x Set the objective magnification Sec 1 2 12 1 x Set the zoom ratio to 1X Sec 1 2 12 2 x Set the channels Sec 1 2 12 3 x Set the highest scan speed Sec 1 2 12 4 x Select the XY observation...

Page 60: ...up period should be more than 10 minutes when the laser system in use uses an Ar laser or more than 30 minutes when it uses a Kr laser or Green HeNe laser x Turning the computer PC ON When the power...

Page 61: ...fan will start After the key is set to ON it takes tens of seconds before the laser oscillation starts Kr laser 1 Set the power switch to ON 2 Turn the key to the ON position The laser fan will start...

Page 62: ...aser oscillation starts LD405 440 laser 1 Confirm that the provided shorting plug is attached to the remote interlock 1 or that the user s equipment is connected to the remote interlock and interlocki...

Page 63: ...30 minutes when it uses a Kr laser Green HeNe laser or HeCd laser x Turning the reflected light power supply unit 1 Set the power switch to ON Once the reflected light power supply unit is turned OFF...

Page 64: ...tialize themselves for about 2 minutes after they are turned on 1 Input the user name and password and start Windows NT or Windows 2000 Input the user name authorized as an Administrator 2 Double clic...

Page 65: ...Getting Started FLUOVIEW Outline of LSM Observation Procedures IV OPERATION INSTRUCTIONS IV 1 2 7 Page 3 The following window appears when the FLUOVIEW software starts Fig 1 3 Window at Start up...

Page 66: ...using the Manual microscope 3 Focus on the specimen by looking into the eyepiece Be sure to adjust the diopter of the eyepiece in advance Refer to the instruction manual of the BX microscope When the...

Page 67: ...1 2 3 2 Combination with IX 1 Turn the light path selector 1 on the right side of the microscope to when using the Manual microscope From the page tabs on the bottom right of the Acquire panel select...

Page 68: ...otor are in use clear the check mark in the Locked check box in the Z Stage sub panel in the Acquire panel see section 2 2 1 3 7 of this volume then focus on the specimen by operating the fine focus a...

Page 69: ...CT3 analyzer 3 is in use disengage it from the light path by setting the switch 5 When only fluorescence observation is required disengage the U DICTHR WI DICTHRA transmitted light DIC 4 by setting th...

Page 70: ...BX61WI LBD FFR This figure shows the case of BX 2 Cube turret Cube display window This figure shows the cube display window 3 Analyzer U MDICT3 4 Transmitted DIC slider U DICTS WI DICTHRA when using B...

Page 71: ...nce observation is required disengage the U DICR transmitted DIC slider 4 by setting the switch to the pulled out position When transmitted light differential interference observation or simultaneous...

Page 72: ...uorescence unit to 4 When the IX2 MDICT analyzer 3 is in use disengage it from the light path by pressing the button 6 on the U HSTR2 hand switch 5 When a fluorescence observation alone is required di...

Page 73: ...ervation Procedures IV OPERATION INSTRUCTIONS IV 1 3 5 Page 1 Fluorescence mirror unit 2 Analyzer IX MDICT Optionally installed 3 Transmitted DIC slider U DICTS optional 5 Filters 4 Condenser 6 Hand s...

Page 74: ...ing the switch to the pulled out position When transmitted light differential interference observation or simultaneous fluorescence transmitted light differential interference observation is required...

Page 75: ...LSM Observation Procedures IV OPERATION INSTRUCTIONS IV 1 3 7 Page 7 Intermediate magnification knob Left side 1 Light path selector 2 Cube turret 4 Transmitted DIC slider U DICT optional 5 Filters 6...

Page 76: ...optimum channels The confocal aperture is also set automatically to the optimum channel for the wavelength and the objective being selected in the Acquire panel Available Dyes list box Lists the avai...

Page 77: ...the Assign dyes manually check box in the Dyes sub panel 2 Select the dyeing method in the Available Dyes list box and drag it directly to the field of the Ch check box 3 After dragging the icon appea...

Page 78: ...ference Examples Dye in Use Detected Channels DETECTION MODE slider FITC Lucifer Yellow etc CH1 Pushed in position mirror FITC TRITC FITC PI etc CH2 TRITC PI CH1 CH2 pushed in half way to the position...

Page 79: ...M488 543 633 1 Mirror 2 SDM570 3 SDM630 1 BA510IF 2 BA530RIF 1 BA560 600 2 BA660IF Ar Kr HeNe R 488 568 633 2 channels 1 DM488 568 633 1 Mirror 2 SDM570 3 SDM630 1 BA510IF 2 BA550RIF 1 BA585 615 2 BA6...

Page 80: ...ctory configuration Up to two types of barrier filter can be equipped per channel If the equipment of another filter set for laser configuration is required please contact your local Olympus represent...

Page 81: ...er intensity by sliding the scale bar in the Laser Intensity group box of the Lasers sub panel in accordance with specimen s brightness fluorescence crosstalk and photo bleaching While using the HeNe...

Page 82: ...item 5 Kr Ar laser line filter switching in section 1 1 1 Scan Unit in this volume 1 2 12 Setting the Observation Condition 1 2 12 1 Setting the Objective Magnification From the drop down list on the...

Page 83: ...g the applicable dyeing method to make the image acquisition ready 2 In the Channel 2 group box check the check box showing the applicable dyeing method to ready the image acquisition To display the i...

Page 84: ...Settings sub panel Select either option button in the Focus Mode group box The focus mode is enabled when acquiring images using the Focus button The Focus function reduces the scanning time by line s...

Page 85: ...lect the XY observation mode option button 1 2 12 6 Repeated Scanning Operation 1 Select the XY Repeat button The acquired image will be displayed in the Live panel Use the FOCUS button to acquire ima...

Page 86: ...ine focus adjustment knob while the Locked check box is checked for this may damage the Z motor 2 While observing the image in the Live panel locate the plane to be observed by displacing the stage us...

Page 87: ...ired for image acquisition When the scan speed is decreased during fluorescence observation the saturation of fluorescence may darken the image of certain types of specimens In this case increase the...

Page 88: ...in the Live panel 2 If no image appears in the Live panel increase the confocal aperture number by 1 using the confocal aperture rotary knob 1 3 While observing the image in the Live panel adjust the...

Page 89: ...Getting Started FLUOVIEW Outline of LSM Observation Procedures IV OPERATION INSTRUCTIONS IV 1 5 1 Page Fig 1 6 Image Acquired in the Live Panel...

Page 90: ...more than one image are saved simultaneously or only one of the images is saved Use the Display channel switch buttons to select the images to be saved The selected images will be saved under the cond...

Page 91: ...button Then this software finishes 3 To logout from this software select the Close all experiments and log in as a different user option button and click the Yes button The dialog box asking whether...

Page 92: ...The fan will stop automatically in a few minutes i e when the laser has cooled down Refer to the instruction manual of your laser unit HeNe Green Red laser Turn the key to the OFF position HeCd laser...

Page 93: ...ure below shows the initial display table of contents of the FLUOVIEW Online Help window To display this window click the Help button in the toolbar at the bottom left of the screen Some words in the...

Page 94: ...ld immediately above the list box 3 Click the Apply button to apply the selected dyeing method to the Channel 1 Channel 2 group box on the upper part of the Acquire panel Available Dyes list box Lists...

Page 95: ...is selected from the Available Dyes list box and the Apply button is clicked the dyeing method will be set automatically to the optimum channels The pinhole diameters are also automatically set to th...

Page 96: ...es manually check box in the Dyes sub panel 2 Select the dyeing method in the Available Dyes list box and drag it directly to the field of the Ch check box 3 After dragging the icon appears on the rig...

Page 97: ...e Help IV OPERATION INSTRUCTIONS IV 1 5 9 Page 1 3 2 1 Configuring the Microscope 1 Display the Acquire panel Settings Z Stage Time Series Dyes Lasers sub panels These are used to set the information...

Page 98: ...cope Configuration Window Scope Control button Displays useful information for the system setup SU group box BARRIER FILTERS CH2 indicates the barrier filters for Channel 2 BARRIER FILTERS CH1 indicat...

Page 99: ...linking does not stop even after the indicated configuration setting has been changed 6 While looking into the microscope move the stage and check the observed image 1 3 2 2 Configuring the Scan Unit...

Page 100: ...light path according to the dyeing method selected for the specimen Use the following procedure to change these filters 1 From the panel page tabs shown on the bottom right of the Acquire panel selec...

Page 101: ...r operation the laser type is set and displayed automatically When the On button is pressed in the laser is oscillating the beam When not using laser for a long time in order to suppress useless elect...

Page 102: ...he combination with BX51 BX61 or IX81 is in use the microscope and scan unit FV500 can be configured on the FLUOVIEW software Use the following procedure to configure the microscope 1 From the panel p...

Page 103: ...oath The top lens is disengaged from the light path Condenser Turret group box Clicking the universal condenser automatically switches the turret Aperture Iris group box Changes the AS value EPI lamp...

Page 104: ...ube automatically switches the turret Shutter group box Clicking inside the box switches the EPI shutter to be closed opened The shutter is opened The shutter is closed EPI lamp Indicates the EPI lamp...

Page 105: ...e group box Checking here disengages the top lens from the light path when the objective of u4 or lower magnification is selected in the Nosepiece group box BX only Checking here closes the FL shutter...

Page 106: ...s shown below When the setting file which you want to read is not displayed in the list box use the Look in drop down list and select the drive or directory where the file is saved Select BX Setting F...

Page 107: ...name text box Select the Save button to close the dialog box z The Focus Setting button enables parfocal corrections and adjustments of jogging sensitivity for the objectives See section 1 3 2 5 Parfo...

Page 108: ...ed To use the setup wizard to begin the parfocality correction from the value of the objective with maximum magnification in descending order click the Next button in the Pfcl Setup Wizard group box O...

Page 109: ...n the parfocality correction value setting with the objective with the maximum magnification click the Switch to option button then click the Next button Click to set the objective with the maximum ma...

Page 110: ...V 1 7 2 Page The objective set as the specified in the Objectives group box is selected and the Pfcl Setup Wizard group box displays the message shown below Bring the specimen in focus by observing it...

Page 111: ...y correction values of objectives with other power values simply click the Next button Click to proceed to the setting of the objective with next higher power to the current objective Click to select...

Page 112: ...ck to set the current focal position as the parfocality correction value for the current objective and proceed to the setting of the next objective Click to set the parfocality correction value of the...

Page 113: ...og Fine drop down list The drop down list appears when clicking each value in the Jog Fine After completing the settings click the Exit button to close the dialog box When a dialog box is displayed to...

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Page 115: ...using the FLUOVIEW system by acquiring an image or opening an image from a file The procedures for the subsequent operations such as image processing are not in question here For the detailed operatio...

Page 116: ...imation display Section 2 9 2 Continuous display Frame by frame display Section 2 9 1 x Observation of image change over time Side by side image display Section 2 10 1 Continuous display Frame by fram...

Page 117: ...n 2 2 2 2 XYZ observation Section 2 2 2 4 XYT observation Section 2 2 2 5 XYZT observation Section 2 2 2 6 TIME Acquire an image in an observation mode FLUOVIEW provides the following observation mode...

Page 118: ...ction 2 2 2 Image Acquisition in Other Observation Modes as well as the following procedure Set the acquisition parameters Section 2 2 1 3 z Observation mode z Scanning speed z Image brightness Acquir...

Page 119: ...reviously acquired image in a file measure the intensity distribution and edit data with Excel Specify the area to be measured Measure the intensity distribution Section 2 7 2 Exit from the FLUOVIEW s...

Page 120: ...from the FLUOVIEW software and turn power OFF Sections 1 2 14 1 2 15 Output the image at the printer Section 2 13 Open an image in a file Section 2 3 2 Exit from the FLUOVIEW software and turn power O...

Page 121: ...N INSTRUCTIONS IV 2 7 Page 2 2 Image Acquisition Confirm the image to be acquired using the microscope and acquire its image using FLUOVIEW The image can be saved in a file as required Do not move FLO...

Page 122: ...in OPERATION Set the channel to be acquired Section 2 2 1 3 3 in OPERATION Set the highest scan speed Section 2 2 1 3 4 in OPERATION Perform repeated scanning Section 2 2 1 3 6 in OPERATION Set the X...

Page 123: ...image acquisition XY Repeat button Repeats XY scanning to display images successively in the Live panel Option buttons Check a button to select one of the displayed observation modes Scan Speed group...

Page 124: ...the Acquire panel Available Dyes list box Lists the available dyes Select the desired items from this list and drag them to the field above it to select the dyeing method Clear button Clear the set dy...

Page 125: ...automatically One Point The Assign dyes manually check box can also be used to set the dyeing method to the desired channel 1 Check the Assign dyes manually check box in the Dyes sub panel 2 Select th...

Page 126: ...age tabs on the bottom right of the Acquire panel select the Settings sub panel Fig 2 4 Settings Sub panel 4 Select the Scope Control button The window as shown below will appear Fig 2 5 Microscope Co...

Page 127: ...nking does not stop even after the indicated configuration setting has been changed 7 While looking into the microscope move the stage and check the observed image 2 2 1 2 Configuring the Scan Unit 1...

Page 128: ...2 Setting the Zoom Ratio to 1X 1 Use the Zoom scale to set the zoom ratio to X1 Fig 2 6 Pan Zoom Group Box 3 Setting the Channels 1 In the Channel 1 group box check the check box showing the applicabl...

Page 129: ...the page tabs on the bottom right of the Acquire panel select the Settings sub panel Select either option button in the Focus Mode group box The focus mode is enabled when acquiring images using the...

Page 130: ...epeated Scanning Operation 1 Select the XY Repeat button The acquired image will be displayed in the Live panel Use the FOCUS button to acquire image at an even higher speed If the specimen is already...

Page 131: ...or detailed operations 1 Check the Locked check box in the Z Stage sub panel Do not turn the fine focus adjustment knob while the Locked check box is checked for this may damage the Z motor 2 While ob...

Page 132: ...blue circle to the left of the Zoom scale represents the field visible through the microscope and the blue frame indicates the acquired image area Move the blue frame inside the circle using the Pan...

Page 133: ...cquisition This value can be set independently from the Offset For example When the acquired image is dark or the observation targets are hardly visible increase PMT voltage to improve the sensitivity...

Page 134: ...lor Tool will appear 2 Click Hi Lo LUT from group box of Standard Color LUTs 3 The intensity value 0 is colored with Blue and the maximum intensity is colored with Red 4 When there is noises in the im...

Page 135: ...llows the acquired image quality to be improved However a low scan speed also lengthens the time required for image acquisition When the scan speed is decreased during fluorescence observation the sat...

Page 136: ...effective for reduction of noise z The Peak accumulation acquires images for the specified number of times while adding the images and stops image acquisition when any intensity value on the image rea...

Page 137: ...de 1 From the page tabs on the bottom right of the Acquire panel select the Settings sub panel Fig 2 9 Settings sub panel 2 In the Filter Mode group box select the Kalman or Accumulate To Peak option...

Page 138: ...The accumulation count can be set up a maximum of 63 times When 0 is set as the number of times of accumulation an ordinary image acquisition is to be performed 4 When Accumulate To Peak is selected e...

Page 139: ...tabs on the bottom right of the Acquire panel select the Settings sub panel Fig 2 10 Settings sub panel 2 In the Filter Mode group box select the Kalman option button 3 Enter the accumulation count i...

Page 140: ...ation In frame mode the image is accumulated every one frame during acquisition In line mode accumulation is done every one line during acquisition The frame mode is intended to utilize for fixed spec...

Page 141: ...tion in this section will be focused on the image acquisition operations in the XZ observation mode that are not used in the XY observation modes which are the operations enclosed in in the chart on t...

Page 142: ...lower scan speed Section 2 2 1 3 10 in OPERATION Re adjust the image brightness Section 2 2 1 3 9 in OPERATION Stop repeated scanning Section 2 2 1 3 11 in OPERATION Set the observation mode Section...

Page 143: ...stop position in the range of the observed cross section Z direction scanning range Start Z text box Shows the scan start position in the range of the observed cross section Z direction scanning range...

Page 144: ...ons 3 When the upper edge position is located click the Set button The Start Z text box will show the scan start position of the range of the multiple sections to be observed Z direction scanning rang...

Page 145: ...select the Z Stage sub panel 2 The number of steps has already been set and displayed in the Step Size text box This number can be changed using the or button in the Step Size text box The number of s...

Page 146: ...APPLIED OPERATIONS Image Acquisition IV OPERATION INSTRUCTIONS IV 2 3 2 Page 5 Acquiring image 1 Click the XZ button in the Acquire panel The acquired image will be displayed in the Live panel...

Page 147: ...2 1 3 1 in OPERATION Set the dyeing method Section 2 2 1 1 in OPERATION Set the channel to be acquired Section 2 2 1 3 3 in OPERATION Set the highest scan speed Section 2 2 1 3 4 in OPERATION Perform...

Page 148: ...2000 times successively at the set scanning speed For reference the time required for the image acquisitions is shown in the frame on the bottom left of the Scan Speed group box The number of image a...

Page 149: ...PERATION Re adjust the image brightness Section 2 2 1 3 9 in OPERATION Stop repeated scanning Section 2 2 1 3 11 in OPERATION Set the observation mode Section 2 2 2 3 2 in OPERATION Acquire image Sect...

Page 150: ...button the move stops Stop Z text box Shows the scan stop position in the range of the observed cross section Z direction scanning range Start Z text box Shows the scan start position in the range of...

Page 151: ...he image in the Live panel locate the bottom edge of the range to be observed by moving up the stage using the Z stage coarse adjustment and Z stage fine adjustment buttons in the Z Stage sub panel Wh...

Page 152: ...ze text box has been calculated by the system so that the depth scale of the acquired image is identical to the horizontal scale The number of steps calculated by the system may be erroneous unless th...

Page 153: ...also sets the Interval text box to Free Run 6 Setting the number of scans 1 Set the number of scans using the or button in the N text box in the Time Series sub panel TIP Rest Time text box Shows the...

Page 154: ...n the XZT observation mode the buttons at the upper part of the Acquire panel changes as shown below Click the Append Next button to append an image An image will be acquired with the same number of s...

Page 155: ...the highest scan speed Section 2 2 1 3 4 in OPERATION Perform repeated scanning Section 2 2 1 3 6 in OPERATION Set the XY observation mode Section 2 2 1 3 5 in OPERATION Set the observation range Sect...

Page 156: ...position in the range of the observed cross section Z direction scanning range Start Z text box Shows the scan start position in the range of the observed cross section Z direction scanning range Set...

Page 157: ...ons 3 When the upper edge position is located click the Set button The Start Z text box will show the scan start position of the range of the multiple sections to be observed Z direction scanning rang...

Page 158: ...tep Size number of steps 3 Setting the observation mode 1 Click the Surface option button in the Mode group box in the Acquire panel 2 In the Acquire panel select the XZT observation mode option butto...

Page 159: ...nges as shown below Click the Append Next button to append an image An image will be acquired with the same number of steps as the image acquired immediately before and appended to it Click the Series...

Page 160: ...eing method Section 2 2 1 1 in OPERATION Set the channel to be acquired Section 2 2 1 3 3 in OPERATION Set the highest scan speed Section 2 2 1 3 4 in OPERATION Perform repeated scanning Section 2 2 1...

Page 161: ...Interval text box An image with no interval can be acquired by entering 0 in the Interval text box In this case the Interval text box shows Free Run message Using button also sets the Interval text b...

Page 162: ...d by heat causing the focusing to be deviated To acquire precise image data it is recommended to set the power switch of the transmitted light illumination to O OFF While acquiring an image in the XYT...

Page 163: ...be focused on the image acquisition operations in the XYZT observation mode that are not used in the XY observation modes which are the operations enclosed in in the chart on the next page For other o...

Page 164: ...n 2 2 1 3 8 in OPERATION Set a lower scan speed Section 2 2 1 3 10 in OPERATION Stop repeated scanning Section 2 2 1 3 11 in OPERATION Set the observation mode Section 2 2 2 6 3 in OPERATION Acquire i...

Page 165: ...ition in the range of the observed cross section Z direction scanning range Start Z text box Shows the scan start position in the range of the observed cross section Z direction scanning range Set but...

Page 166: ...ons 3 When the upper edge position is located click the Set button The Start Z text box will show the scan start position of the range of the multiple sections to be observed Z direction scanning rang...

Page 167: ...l to the horizontal scale The number of acquired images shown in the Slices text box can also be input from the keyboard After setting Start Z Z direction scan start position and Stop Z Z direction sc...

Page 168: ...the Interval text box to Free Run 5 Setting the number of scans 1 Set the number of scans using the or button in the N text box in the Time Series sub panel Fig 2 17 Time Series Sub panel TIP N text b...

Page 169: ...acquire precise image data it is recommended to turn off the power of the transmitted light While acquiring an image in the XYZT observation mode clicking the STOP SCAN button changes the buttons at...

Page 170: ...low 570 nm the image is acquired in channel 1 Ch1 When the specimen is dyed so that the wavelength of fluorescence emission from it is above 570 nm the image is acquired in channel 2 Ch2 z Set the DET...

Page 171: ...he dyeing method emitting a shorter light wavelength than 570 nm is set to Ch1 and that emitting a longer light wavelength is set to Ch2 automatically Available Dyes list box Lists the available dyes...

Page 172: ...es manually check box in the Dyes sub panel 2 Select the dyeing method in the Available Dyes list box and drag it directly to the field of the Ch check box 3 After dragging the icon appears on the rig...

Page 173: ...group box that shows the dyeing method is check marked to indicate that the corresponding channel is ready for image acquisition z Set the image brightness Adjust the brightness of the acquisition cha...

Page 174: ...ng methods in the Available Dyes list box in the Selected Dyes group box and drag the selected method into the field immediately above the list box 3 Click the Apply button to apply the selected dyein...

Page 175: ...h2 automatically One Point The Assign dyes manually check box can also be used to set the dyeing method to the desired channel 1 Check the Assign dyes manually check box in the Dyes sub panel 2 Select...

Page 176: ...ow z Make the two channels ready for image acquisition In the Acquire panel make sure that the check box in the Ch1 and Ch2 group boxes that show the dyeing methods are check marked to indicate that t...

Page 177: ...cquire panel select the Dyes sub panel Fig 2 20 Dyes Sub panel 2 Select each of the desired specimen dyeing methods in the Available Dyes list box in the Selected Dyes group box and drag the selected...

Page 178: ...h2 automatically One Point The Assign dyes manually check box can also be used to set the dyeing method to the desired channel 1 Check the Assign dyes manually check box in the Dyes sub panel 2 Select...

Page 179: ...e instructions given by the Microscope Configuration window z Turn off the transmitted light bulb of the microscope 1 From the page tabs on the bottom right of the Acquire panel select the Settings su...

Page 180: ...g channel is ready for image acquisition z Set the brightness of the transmitted image Adjust the brightness of the image of Ch3 image by using the PMT Offset and Gain LED sliders in the Ch3 group box...

Page 181: ...ion Mode 2 2 2 Image Acquisition in Other Observation Modes 2 2 5 Image Acquisition of Only the Rectangular Position Clip Scan 2 2 6 Image Acquisition by Magnifying the Rectangular Position Zoom In Sc...

Page 182: ...n area is limited to an area with 400 x 300 pixels halved in both X and Y directions When the image is acquired in this mode the size per pixel on the specimen is identical to that before start of thi...

Page 183: ...at is compatible with rotation scan FV5 IO3 specify the image rotation angle using the rotation arrow buttons on both sides of the circuit in the Pan group box in the Acquire panel 8 Acquire images by...

Page 184: ...be observed and acquires the image of that area by magnifying it For example let us assume that an 800 x 600 pixel image is displayed in the Live panel and the image acquisition area is limited to an...

Page 185: ...4 Change the frame size Place the mouse pointer inside the frame and click to display square handles around the frame Then place the mouse pointer on one of the handles and drag it to change the fram...

Page 186: ...arrow buttons on both sides of the circuit in the Pan group box in the Acquire panel 9 Acquire the image by clicking XYZ XYT XYZT etc 10 After image acquisition select Normal under the Surface XY opti...

Page 187: ...e Mode group box in the Acquire panel and select Fast from the list displayed below it 512 by 512 is set automatically in the Size drop down list in the Mode group box X2 is set and shown in gray out...

Page 188: ...the Acquire panel and select Sequen from the list displayed below it Selecting Sq Clp Sq Rct or Line Sq enables the image acquisition of the desired rectangle area that of the rectangle area at desire...

Page 189: ...tion starts with the smallest group number set To prevent unwanted color fading it is recommended to acquire the image from the reagents with longer wavelengths 5 Click the Seq Once button to acquire...

Page 190: ...Off button to the Rdy button displayed in the pressed in condition 8 Click the OK button in the FLUOVIEW dialog box Image acquisition starts 9 Adjust the brightness with the PMT Offset and Gain LED s...

Page 191: ...rs according to the preset condition by increasing the number of detection channels With the FV300 they should be switched manually Operation Example The following procedure deals with the case of obs...

Page 192: ...ion mode is set to Sequential Scan the Dyes sub panel in the Acquire panel cannot be used 4 Using the and buttons in Group set the group according to the reagent in use This operation is not required...

Page 193: ...the physically identical channels an ordinary channel and virtual channel in the same group For example ordinary channel 1 and virtual channel 1 cannot be set in the same group Sequential Group means...

Page 194: ...in the Virtual Channel group box and select 2 In the Dye drop down list of vCH 2 select CY5 Displayed each optimum filters etc in Excitation DM Beam Splitter Barrier Filter and Laser Unit group box A...

Page 195: ...oup box 7 Adjust the image brightness for every group Select the group to adjust the image brightness in the Group drop down list displayed on the upper part of the Acquire panel Select a group before...

Page 196: ...scan for Group at Optical System Configuration window so that this dialog does not appear and scanning can be executed consecutively 14 After image acquisition click Normal below the Surface XY option...

Page 197: ...ears in the Live panel Slant under the Depth XZ option button is selected in the following Fig 3 Move the line to the area you want to observe To move the line place the mouse pointer on it and drag t...

Page 198: ...ick the mouse left button to fix the line inclination angle 7 Set the interval time or the number of scans The operation procedure is same as that in the XY observation mode See section 2 2 2 XT Obser...

Page 199: ...ating the laser to certain point on the image If you have FLUOVIEW TIME COURSE software image acquisition can be started with buttons keys or input of external trigger signals 1 Acquire an image in th...

Page 200: ...ty values appears in the Live panel 3 Move the cross cursor to the area you want to observe To move the cross cursor place the mouse pointer on it and drag the mouse 4 Set the scan speed for image acq...

Page 201: ...page tabs on the bottom right of the Acuire panel select the Time Series sub panel Set the time for measurement with the or button in the Scanning text box 6 Set the starting method for image acquisi...

Page 202: ...size of the graph window can be modified by double clicking the mouse button on it When a certain area is specified by dragging the mouse from the top left to the bottom right on the graph while pres...

Page 203: ...bservation When the specimen is in curved form the image can be acquired by drawing a desired line on it 1 Acquire an image in the XY observation mode For detailed operation see section 2 2 1 Image Ac...

Page 204: ...ged on the handle The handle can be add by clicking the mouse on desired coordinates To delete the handle click the mouse on it Right click the mouse on the coordinates but the handles to fix the shap...

Page 205: ...ick XZ XT or XZT etc to acquire the image The image shown in the Live panel after image acquisition is the image that acquired at the curved line specified on the image and arranged uprightly in the X...

Page 206: ...excitation in this section Three kinds of the laser excitation mode Disable REX and Bleach are selectable according to your purpose And each mode can be switched in the XYT observation In use of the R...

Page 207: ...image acquisition in the Line mode Normal Slant and Free can not be performed The specimen in the images carried in this section is not for FRAP but for the user s manual 2 2 12 1 Making REX Mask Fil...

Page 208: ...ngular Circle Polyregion or Free region button And drag on the image to specify the region of laser excitation In order to specify two or more regions click the mouse on or inside the region with hold...

Page 209: ...ers sub panel Fig 2 24 Lasers sub panel When the Make REX mask button is displayed in gray click and select the specified region on the Live panel When two or more regions are specified select all the...

Page 210: ...e REX Live panel has already been created on the FV300 software A panel is continuously created after REX_Live0 The REX mask file can be saved The saving procedure is completely the same as that of a...

Page 211: ...set in the Mask Intensity dialog box The laser intensity of the REX mask file is setup on percentage to 100 The region becomes brighter as the value is close to 100 In case of using REX function setti...

Page 212: ...It is because the square power of zoom magnification per time can be secured Moreover using the objective of larger NA concentrates the laser irradiation spot in Z direction and stronger laser can be...

Page 213: ...mple after acquiring an image 3 Making REX Mask File Refer to section 2 2 12 1 Making REX Mask File for the procedure to make the REX mask file If the setup of the region of laser excitation remains a...

Page 214: ...e mouse inside the frame to display the pop up menu as shown below Select the image to be masked in the menu In order to use the already saved file as a REX mask file open the image beforehand The ope...

Page 215: ...EX mask file is completely the same as that of an image For details refer to section 2 3 2 Opening Previously Saved Images in this manual 6 The icon of the selected REX mask file is displayed inside t...

Page 216: ...ge Acquisition IV OPERATION INSTRUCTIONS IV 2 1 0 2 Page 5 Setting the XYT Observation mode 1 Confirm that XY Norm is displayed in the Mode group box in the Acquire panel 2 Select the XYT option butto...

Page 217: ...the Total Time text box The time for image acquisition differs according to the interval and the number of scans 6 Setting the TIME COURSE Software optional Setting of the real time graph is required...

Page 218: ...ver the REX mode is effective to prevent fading of the obtained data 1 Select the Disabled option button in the AOTF group box in the Lasers sub panel 2 Click the XYT button to start image acquisition...

Page 219: ...cted Select the Disabled or REX mode option button within 10 seconds before the next scanning is started In the TIME COURSE software optional when the REX mask file in the REX or Bleach mode is change...

Page 220: ...specimen is acquired When performing the forth or subsequent scan be sure to switch the REX mode and Bleach mode between scans In the TIME COURSE software optional when the REX mask file in the REX o...

Page 221: ...mation described below when scanning is executed and the information appears in the same state when each scan mode is selected next time Angle Profile Position When default setting information is desi...

Page 222: ...can O O Clip scan O O Zoom in scan O O Fast normal scan O O Fast clip scan O O Sequential normal scan O O Sequential clip scan O O Sequential zoom in scan O O Line sequential normal scan O O Line sequ...

Page 223: ...Fast normal scan O O Fast clip scan X O Line sequential normal scan O O Line sequential clip scan X O XT Line sequential zoom in scan X O Normal scan O O Clip scan X O Zoom in scan O O Free line scan...

Page 224: ...s used with the following method In case of I O2 the arrow of Pan Zoom group box of Acquire panel is grayed out and cannot be selected when it boots up as shown below In case of I O3 the arrow of Pan...

Page 225: ...not displayed double click the hierarchical level above the current level Experiment button Saves the image displayed in the Display panel together with its file name Display button Saves the image di...

Page 226: ...s are also displayed in the Icon field in the Files list box In all observation modes the icons for 3 or more channels are identical Image Icon Significance XZ observation XZ observation 2 channel mod...

Page 227: ...names Icon Icons showing the image observation modes Y Image sizes in the Y direction X Image sizes in the X direction T Number of images acquired over time Ch Number of image channels Date Image sav...

Page 228: ...t position button Also set the save end position in the similar way 3 When the images to be saved are acquired from more than one channel select whether saving images from more than one channel simult...

Page 229: ...appears in File Name text box Then the image file can store with the numbering just pressing Save button 8 Click the Save button If a file with the same name as the entered file name already exists a...

Page 230: ...it is required to use a FLUOVIEW image in another application 1 Display the Display panel showing the image to be saved at the front 2 When there is more than one image to be saved display the image...

Page 231: ...APPLIED OPERATIONS Saving Opening and Shredding Images IV OPERATION INSTRUCTIONS IV 2 1 1 7 Page Fig 2 29 Save Experiment As Dialog Box...

Page 232: ...he file name in the File Name text box 9 Click the Save button If a file with the same name as the entered file name already exists a dialog box is displayed to ask if you want to overwrite the existi...

Page 233: ...ltiple channels are saved together or the image slice of only 1 channel is saved Use the display switching button for the selection The image will be saved in the channel condition as displayed For th...

Page 234: ...n so that the list of buttons disappear 7 Click the Experiment button in the Save group box The Save Experiment As dialog box appears Fig 2 30 Save Experiment As Dialog Box 8 Use the Save In drop down...

Page 235: ...2 1 Page 12 A dialog box as shown below appears Fig 2 31 Saving a subset of Experiment s Data Dialog Box The Saving a subset of Experiment s Data dialog box does not appear if no area is specified in...

Page 236: ...he Successive Display Speed One Point When saving image slices acquired in Z observation it is recommended to lower the animation display mode by clicking the Turtle button If image slices are saved w...

Page 237: ...hen it is required to change the saved file type use the Save as Type drop down list Two saved file types can be selected AVI Compressed Saves the AVI file being compressed AVI Uncompressed Saves the...

Page 238: ...if x Single TIF s 16 bit tif TIFF Tagged Image File Format is used for image x Single TIF s 24 bit tif exchange between applications or computers Three types including the 8 bit 16 bit and 24 bit type...

Page 239: ...ct when the image will be analyzed or processed after save Opening the image also opens the comment which can be changed or measured as required Single TIF s Select when the image will be handed to an...

Page 240: ...ving extended image Z T series switch button Comment drawn on images Fluoview Multi Tiff The image of only Ch3 is saved Save file name abcdefgh tif The images of Ch1 Ch2 and Ch3 are saved Save file na...

Page 241: ...ames abcdefgh0 tif Ch1 No 0 abcdefgh3 tif Ch1 No 3 abcdefgh4 tif Ch2 No 0 abcdefgh7 tif Ch2 No 3 abcdefgh8 tif Ch3 No 0 abcdefgh11 tif Ch3 No 3 4 images are saved only from Ch3 Save file names abcdefg...

Page 242: ...gh3 tif Ch1 No 3 abcdefgh4 tif Ch2 No 0 abcdefgh7 tif Ch2 No 3 abcdefgh8 tif Ch3 No 0 abcdefgh11 tif Ch3 No 3 4 images are saved only from Ch3 Save file names abcdefgh0 tif Ch3 No 0 abcdefgh3 tif Ch3...

Page 243: ...efghV5 tif Ch3 No 1 abcdefghV6 tif Ch1 No 2 abcdefghV8 tif Ch3 No 2 abcdefghV9 tif Ch1 No 3 abcdefghV11 tif Ch3 No 3 4 images are saved only from Ch3 Save file names abcdefgh0V0 tif Ch1 No 0 abcdefgh0...

Page 244: ...s abcdefgh0 bmp Ch1 No 0 abcdefgh3 bmp Ch1 No 3 abcdefgh4 bmp Ch2 No 0 abcdefgh7 bmp Ch2 No 3 abcdefgh8 bmp Ch3 No 0 abcdefgh11 bmp Ch3 No 3 12 images are saved from each of Ch1 Ch2 and Ch3 Save file...

Page 245: ...5 bmp Ch3 No 1 abcdefghV6 bmp Ch1 No 2 abcdefghV8 bmp Ch3 No 2 abcdefghV9 bmp Ch1 No 3 abcdefghV11 bmp Ch3 No 3 4 images are saved only from Ch3 Save file names abcdefgh0V0 bmp Ch1 No 0 abcdefgh0V3 bm...

Page 246: ...From the File Type drop down list select the file type of the files to be listed in the Files list box 3 Click the Experiment button in the Load group box Other methods are also available for opening...

Page 247: ...tom of the File I O panel The Experiments in Memory dialog box appears as shown below Fig 2 33 Experiments in Memory Dialog Box 2 In the Experiments in Memory dialog box select the file name of the im...

Page 248: ...can be shredded simultaneously 1 Display the Experiments in Memory dialog box while showing multiple images in the Display panel 2 Pressing down the Shift key select multiple image files to be shredde...

Page 249: ...the File I O panel The Experiments in Memory dialog box appears as shown below Fig 2 34 Experiments in Memory Dialog Box 2 In the Experiments in Memory dialog box select the file name of the image to...

Page 250: ...in the Comments panel is saved simply but it is not yet saved in the image file To save the comment in the image file use the Save button in the File I O panel 6 Click the OK button 7 Click the Done...

Page 251: ...quisition parameters and click the Comments button 3 Display the Image Attributes panel at the front The panel as shown below appears where the information on the selected image is shown Fig 2 37 Imag...

Page 252: ...ck the Set Attributes button During modification of the lengths per image pixel the number of steps in the Z direction or the objective setting if it is required to restore the previous setting click...

Page 253: ...ge Fields Panel 5 Select the information items to be checked from the list box on the right The kind of information displayed when each item is selected is shown below 6 After checking the information...

Page 254: ...1 4 0 Page One Point The Image Comments dialog box can also be displayed by a mouse operation 1 Display the image to be saved at the front of the Display panel and right click a point in the image 2 A...

Page 255: ...tion and observation condition can be saved as ASCII text file in a disk 1 Display the Display panel of the image to be saved at the front position 2 Select the Experiment List button in the tool bar...

Page 256: ...pears Fig 2 41 Save to txt dialog box The LUT Ch1 to LUT Ch6 check boxes are displayed according to the channel used for image acquisition 5 Check the check box of the item to be saved Properties butt...

Page 257: ...irm that ASCII text txt is selected in the Save as type drop down list 9 Enter a file name into the File Name text box 10 Click the Save button When the text file having the same name as the entered n...

Page 258: ...egion can be saved in a file and they can also be read 2 3 7 1 Saving the Region File 1 Display the Display panel of the image including the region to be saved at the front position 2 Select the Exper...

Page 259: ...omments dialog box 4 Select the Annotations ROIs button in the Export to File group box The Save Experiment As dialog box as shown below appears Fig 2 45 Save Experiment As dialog box 5 Select the sav...

Page 260: ...d enter another name to save the file 2 3 7 2 Reading the Region File 1 Display the Display panel of the image including the region to be read at the front position 2 Select the Experiment List button...

Page 261: ...iment As dialog box 5 Select the drive or folder where the file to be read is saved in the Look in drop down list 6 In the list below the Look in drop down list double click the folder or sub folder w...

Page 262: ...size of the file including the region to be read and that of the specified file differ the Careful dialog box as shown below appears Clicking the OK button reads the file in different image size where...

Page 263: ...setting The protocol can save and modify to be suitable to new experiment Refer to the reference sections written in the following diagram for detail operation Start the Protocol Processor Section 2...

Page 264: ...quisition Protocol Processor window appears as shown below It is referred to as the PAPP window in this manual PAPP window appears where is the last time in use Start button Starts the protocol Break...

Page 265: ...be pressed during PAPP protocol is working 2 4 2 Editing the Protocol The protocol of the Protocol Processor consists of several Tracks and Track is a set of image acquisition condition and parameter...

Page 266: ...automatically be adjusted according to the length of characters Get Init Values button Click to acquire the current settings in the Acquire panel as the initial values of the settings Init Val box Sh...

Page 267: ...mode This mode can set the previous scan mode to acquire consecutive XYT XYZT or XZT image with the acquisition parameter modified Sub menu of New mode Description Series XY Acquires XY image X XYT A...

Page 268: ...ew mode Change Input details Condition column of the following Series Mode XY X Set 1 automatically XYT O Set number of images to be acquired Enter number of slices to acquire XYZ X Set 1 automaticall...

Page 269: ...cessing Angle Assign rotated image scan angle of field of view 0 through 359 can be entered Angle can not set at a certain sub scan mode and a certain hardware Please see 2 4 10 Restrictions at PAPP w...

Page 270: ...ting Procedure of XYT Observation for hours Protocol using repetition processing Scan speed image acquisition speed from Fast Middle Slow and Init in the menu The above speed is the same with those of...

Page 271: ...tage Track No When multi point time lapse software is used track number of multi point time lapse software that meets to this track will be displayed When multi point time lapse software is not used t...

Page 272: ...00 01 10 and 11 in the pull down menu Comment Enter any comment as desired In case of the range of acceptable values are specified for a cell it is checked according to the specified range If a value...

Page 273: ...appended image data type from T series image Z series image or animation image The default is T series Date Name Specify the order of append date or name The default is date CopyROI CopyROI Copies act...

Page 274: ...case that NORMAL is selected no setting is required E g In case that integration is required 4 times using KALMAN filter enter Filter KALMAN 4 Gain Gain ch value Sets the Gain value of channel E g En...

Page 275: ...tails see 2 4 7 Save Save Saves whole series of images SendCOM SendCOM message 0 1 Sends data through RS 232C to other PC The following data has to be returned in synchronous mode Completed normally O...

Page 276: ...displayed on Zstage sub panel of Acquire panel of FV300 500 may occur Use the handle after confirming the moving range adequately In case that Z stage is controlled with use of command lock Z motor D...

Page 277: ...stination of file save using the Save and Stop Save commands is the path displayed on the left of the Image File Name text box If it is required to change the file save destination click the Path butt...

Page 278: ...les start To end Step n Define a variable name in Variables the start value in start end value in End and an integer representing the number of steps in n The range to which the variable is specified...

Page 279: ...ols that acquire image with interval of step 10 by N equal to 10 to 100 Enter N 10 To 100 Step 10 for Condition column in No 1 track Define laser intensity of argon laser as variable N for No 2 track...

Page 280: ...the setting of item checked at check box of group box name is set to FLUOVIEW software x Set Init Val button loads Init Val box in PAPP window into Track parameters One Point It is possible to select...

Page 281: ...idth when the window was previously closed x Left side of Protocol Creator window has Frame tab Command tab and Loop tab Track can be made by drag and drop those icon s into Track Image Track window a...

Page 282: ...button the detail information is displayed By pressing Less button close the information display During Protocol Creator window is displayed PAPP window can not be operated 2 4 3 Saving the Protocol...

Page 283: ...elect file type pap or csv with Save as type dropdown list 5 Click the Save button If a CSV file having the same file name as the file name entered above already exists a dialog box appears to ask if...

Page 284: ...tocol Before executing a protocol check the To Bottom on Running in the View menu in the PAPP dialog box x Shrink the PAPP dialog box at any position in the screen while executing a protocol Before ex...

Page 285: ...d the protocol Select Skip in the Run menu in the PAPP window x Stop executing the protocol Select End in the Run menu in the PAPP window One Point When Stop when error occurs is selected checked from...

Page 286: ...l in it 1 Start the Protocol Processor software and display the PAPP window For details see section 2 4 1 Starting the Protocol Processor 2 In the File menu select Open The File Open dialog box appear...

Page 287: ...d to XYT or XYZT in count of Z slice parameter If Z Slices 1 the Frame becomes XYT If Z Slices is more than 1 the Frame becomes XYZT If the Frame is continuous one Frame becomes Series Fig 2 51 Sample...

Page 288: ...ssor IV OPERATION INSTRUCTIONS IV 2 1 7 4 Page Fig 2 52 Multi Point Time Lapse Protocol Rule C x For and Command in Series appears with indented x Combined For Next structure becomes single layer Fig...

Page 289: ...dropdown list x By clicking Set button to be able to use the port COM Control dialog box x SendCOM and ReceiveCOM commands are available when Use RS232 C command in protocol in Option menu is checked...

Page 290: ...c Initial image acquisition Track 1 sets the zoom ratio to 2 x and acquires an image Photobleaching Track 2 sets the zoom ratio to 10 x and acquires image 3 times to make photobleach on the specimen R...

Page 291: ...APPLIED OPERATIONS Protocol processor IV OPERATION INSTRUCTIONS IV 2 1 7 7 Page Fig 2 54 Panel After a Repeated Scan...

Page 292: ...the image to display the real time graph For specifying method refer to section 2 4 3 Specifying the Regions Where Intensity Graph is Displayed in Real Time 2 Display the Tiempo sub panel in the Acqu...

Page 293: ...art and edit the protocol after setting of the observation condition has been completed For starting method refer to section 2 4 1 Starting the Protocol Processor Fig 2 56 Panel Displaying the PAPP wi...

Page 294: ...to 10X and acquire three images Irradiate strong laser onto the region to bleach 1 Select Series mode in Mode of Track 2 Select Normal in Start by of Track 3 Enter 3 to Condition of Track 4 Enter 0 t...

Page 295: ...to Condition of Track 4 Enter 5 to Frame Interval 5 Enter 2 to Zoom 6 Select Middle in Scan Speed Return to the setting before bleaching 7 Enter the laser intensity value for image acquisition to Las...

Page 296: ...CTIONS IV 2 1 8 2 Page z z Wait 5 sec Set Zoom power to x2 Set Zoom power to x10 Get 5 images bleach Set Zoom power to x2 Get one image Acquire image Save image Protocol Track 1 Track 2 Track 3 Repeat...

Page 297: ...be saved after being executed 1 Select Save or Save as in the File menu in the PAPP window Save using the dialog box displayed For details see section 0 1 Executing the Protocol Executes the protocol...

Page 298: ...ED OPERATIONS Protocol processor IV OPERATION INSTRUCTIONS IV 2 1 8 4 Page 1 Exiting the Protocol Terminate the protocol after acquiring the images Fig 2 59 Panel When Exiting the Protocol The images...

Page 299: ...Start of repetition Track 2 End of repetition Turnover Track 4 7 repeats Reference image acquisition Acquire 9 images with the zoom power 2 x then save the images acquired in Track 1 Repeated Protocol...

Page 300: ...he Ar laser is variable N and create a protocol for acquiring image as N varies from 10 to 100 in steps of 10 5 In Condition column to track No 1 enter For statement N 10 To 100 Step 10 6 In track No...

Page 301: ...ion Before setting and starting the XYT observation for hours using the Protocol Processor execute a repeated scan and set the condition for the observation Assuming FITC to Channel 2 and image size t...

Page 302: ...art and edit the protocol after setting of the observation condition has been completed For starting method refer to section 2 4 1 Starting the Protocol Processor Fig 2 61 Panel Displaying the PAPP wi...

Page 303: ...acquisition order number Assign the image file name in the Image File Name text box area before protocol starts z Setting Track 2 Specifies N variable and designate range to be repeated Protocol conti...

Page 304: ...of each channel in the cell for each channel in PMT Voltage Enter Init in PMT2 and PMT3 7 Enter the image file name to be stored to Save FileName z Setting Track 4 Specifies the repeated protocol by u...

Page 305: ...he Protocol Save the protocol edited The protocol can also be saved after being executed 1 Select Save or Save as in the File menu in the PAPP window Save using the dialog box displayed For details se...

Page 306: ...ONS IV 2 1 9 2 Page 2 Exiting the Protocol After the initial image has been acquired and saved saving the image acquired in Track 4 in Track 5 is repeated for 7 times after which the protocol terminat...

Page 307: ...cell Select Copy in the Edit menu in the PAPP window x Cut the value in cell Select Cut in the Edit menu in the PAPP window x Paste the value copied Select Paste in the Edit menu in the PAPP window x...

Page 308: ...oomIn O O O O Fast O O X O FastClip O O X O Seq_Normal O O X O Seq_Clip O O X O Seq_ZoomIn O O O O LineSeq_Normal O O X O LineSeq_Clip O O X O XY LineSeq_ZoomIn X O X O Normal O O X O Clip O O X O Zoo...

Page 309: ...X O ZoomIn O O O O FreeLine O O X O Fast O O X O FastClip X O X O LineSeq_Normal O O O O LineSeq_Clip X O X O XT LineSeq_ZoomIn X O X O Normal O O X O Clip X O X O ZoomIn O O O O FreeLine O O X O Lin...

Page 310: ...thod In case that I O2 is used Pan Zoom group boxes on Acquire panel cannot be selected because arrows are grayed out as shown below when the board is started In case that I O3 is used it is possible...

Page 311: ...eq_ZoomIn O O O O Normal X X X X Clip O O O O ZoomIn O O O O Fast X X X X FastClip O O O O Seq_Normal X X X X Seq_Clip O O O O Seq_ZoomIn O O O O LineSeq_Normal X X X X LineSeq_Clip O O O O XYT LineSe...

Page 312: ...O O LineSeq_ZoomIn O O O O Normal X O X O Clip O O O O ZoomIn O O O O FreeLine X X X X LineSeq_Normal X O X O LineSeq_Clip O O O O XZ LineSeq_ZoomIn O O O O Normal X O X O Clip O O O O ZoomIn O O O O...

Page 313: ...When the image was acquired from more than one channel select the channels to be colored using the Ch1 Ch2 and or Ch3 option buttons The Ch1 Ch2 and Ch3 option buttons are displayed only when an imag...

Page 314: ...Color Tool dialog box can also be displayed by a mouse operation 1 Display the image to be colored at the front of the Display panel and right click a point in the image 2 A pop up menu as shown below...

Page 315: ...ng the check boxes below the graph in the Color LUT Tool group box 5 Set the range of intensity graph application using the Low and High scales in the Intensity Mapping group box When the Low scale Hi...

Page 316: ...Finally click the OK button to exit from the LUT editing One Point The Color Tool dialog box can also be displayed by a mouse operation 1 Display the image to be colored at the front of the Display p...

Page 317: ...Intensity Mapping group box Dragging one end of the graph makes it possible to change the inclination The set intensity graph is immediately reflected in the image in the Display panel When the Low sc...

Page 318: ...Display panel of for the image obtained from multiple channels at the front 2 Click the image to display the Display channel switch buttons on the bottom left of the image 3 Click the Channel 1 Channe...

Page 319: ...nnels Use the buttons displayed at the top of the Display panel and those on the bottom right which are displayed when the corresponding image is clicked For the display of the image of only one chann...

Page 320: ...s click one of the Side by side views button or Over And Under views button The icon shown in the Display switching button will change to the icons of the Side by side views button or Over and under v...

Page 321: ...At the same time the icon shown in the Display switching button will change to the icon of the Single View button 4 Click the image to display the Display channel switch buttons at the bottom left of...

Page 322: ...wed simultaneously can be changed Increased image is only to be displayed The image increased in Add View is not subjected to these operations described below 2 5 5 1 Increasing the Number of Divided...

Page 323: ...od IV OPERATION INSTRUCTIONS IV 2 2 0 9 Page 3 Select Views then select Add a view in the sub menu 4 A view is added to the rightmost position on the Display panel Fig 2 71 Display Panel After View Ad...

Page 324: ...IV 2 2 1 0 Page 2 5 5 2 Decreasing the Number of Divided Images 1 Display the Display panel of the image to be changed at the front Fig 2 72 Display panel 2 Right click the image A pop up menu as sho...

Page 325: ...APPLIED OPERATIONS Changing the Image Display Method IV OPERATION INSTRUCTIONS IV 2 2 1 1 Page 4 A view is removed from the Display panel Fig 2 73 Display panel after View Removal...

Page 326: ...slices of image from the start position to the end position sequentially successive display or one frame by frame display or to stop successive display Holding the button displays image slices in suc...

Page 327: ...ox Shows how the images are displayed in the Display panel Columns text box Sets the number of columns or the number of image slices displayed in a horizontal row Rows text box Sets the number of rows...

Page 328: ...the images will be arranged can be confirmed in the gray box at the upper part of the Tiling group box 3 When there are multiple images to be displayed select the following items in the Tile Over drop...

Page 329: ...displayed together for simultaneous view The operation method is identical to the method for displaying images per channel except for the following point See section 2 5 7 1 Displaying Multiple Images...

Page 330: ...s in the Columns and Rows text boxes How the images will be arranged can be confirmed in the gray box at the upper part of the Tiling group box 3 Select T from the Tile Over drop down list 4 When the...

Page 331: ...5 7 5 Displaying Same Images in Different Display Methods Images composed of multiple image slices can be displayed in more than one display methods together 1 Display the Display panel of one of the...

Page 332: ...k the top left image of the displayed images Buttons appear above the clicked image 6 Switch the image display using the displayed buttons For details on the display switching method see sections 2 5...

Page 333: ...Click the Retile or New Page button All of the images in the panel are re displayed using the same display method as that set in step 2 above 2 5 7 7 Displaying Different Images Together Two complete...

Page 334: ...arameters used in the image acquisition are displayed in the Experiment panel The mouse pointer turns into the image icon during dragging 4 Click the Done button in the Experiments in Memory dialog bo...

Page 335: ...play panel appears showing the two images one above the other The image of the file displayed in the frame at the top left of the Tile panel is displayed on the upper part of the Display panel and tha...

Page 336: ...Display the Display panel of the image to be displayed 2 Click the Full Screen button in the toolbar at the bottom left of the screen When the mouse left button is clicked while the image is displaye...

Page 337: ...Page One Point The image can also be displayed full screen by the mouse operation on the image 1 Display the Display panel of the image to be displayed full screen and click the mouse right button on...

Page 338: ...ons as shown below are displayed on the top of the Display panel Usually the Auto button is displayed and clicking it displays the list of buttons shown below Us these buttons to magnify or reduce the...

Page 339: ...ly filtering to images 1 Display the Filters sub panel in the Process panel at the front Fig 2 84 Filters Sub panel Icon of the image being displayed Image to be processed by filtering Smoothing Filte...

Page 340: ...d of multiple image slices the range of image slices to be filtered can be specified using the Set start position and Set end position buttons above the image First display the image slice to start fi...

Page 341: ...the image grains If the image contains noise the noise is also enhanced It has two filter formats X and Y as shown below The format providing the larger value after filtering is used 1 2 1 1 0 1 0 0...

Page 342: ...Laplacian filtering except for the following point See section 2 6 1 1 1 Laplacian filter z Click the Highpass X button 4 High pass Y filter The HIGH PASS Y filter passes the high frequency structure...

Page 343: ...2 6 1 2 Noise Reduction When random noise interferes with an image its irregularity increases and it become harder to see Such noise can be reduced by means of filtering Three kinds of filters are ava...

Page 344: ...e low frequency structures In this way it can eliminate small grains and provide smooth noise reduced image filter format is as shown below 1 2 1 2 4 2 1 2 1 The operation method is identical to Lapla...

Page 345: ...that the correction is possible only with the transmitted images 1 Display the Filters sub panel in the Process panel Fig 2 86 Filters Sub panel 2 Enter the appropriate correction factor for the imag...

Page 346: ...s newly created in the Display panel and shows an image after the correction of the DIC level irregularities of the background Fig 2 87 Level Panel After DIC Level Correction 4 If the DIC level irregu...

Page 347: ...facility changes the contrast by noticing a certain section between 0 and 4095 and mapping this section to values between 0 and 255 1 Display the Process panel at the front 2 Display the Histogram su...

Page 348: ...To select the target channel s use the Display channel switch buttons The histogram of the selected channel s is displayed Example When only the Ch1 image is displayed the histogram of Ch1 is display...

Page 349: ...e multi channel mode select the channel s to be subjected to LUT intensity mapping re assignment using the Channel option buttons 9 Select the region to be subjected to mapping re assignment using the...

Page 350: ...ribed below 1 Display the Math sub panel of the Process panel Fig 2 91 Math Sub panel Scalar Operations group box Provides the command buttons for use in operations between image and constant Sets the...

Page 351: ...the Process panel Before the dragging and dropping the frame at the top left shows the icon of the image file displayed in the Display panel The mouse pointer turns into the image icon during draggin...

Page 352: ...image Click the Add 2 Images button in the Multi Image Operations group box A new Display panel showing Image Image in the page tab appears displaying the image obtained by the addition operation To...

Page 353: ...ion of an image by an image image by a constant is possible as described below The operation method is identical to Image Image Image Constant except for the following point See section 2 6 3 1 Image...

Page 354: ...Image Const in the page tab appears displaying the image obtained by the division operation 2 6 3 5 NOT Image The NOT operation of an image allows the bright and dark areas of the image to be reverse...

Page 355: ...age Processing IV OPERATION INSTRUCTIONS IV 2 2 4 1 Page 3 Click the NOT Image button A new Display panel showing NOT in the page tab appears showing the image obtained by the operation Fig 2 95 NOT P...

Page 356: ...the bottom of the Process panel The Experiments in Memory dialog box appears as shown below Fig 2 97 Experiments in Memory Dialog Box Experiment List button Image AND Image button Executes an AND ope...

Page 357: ...pping the frame at the top left shows the icon of the image file displayed in the Display panel The mouse pointer turns into the image icon during dragging 4 From the Experiments in Memory dialog box...

Page 358: ...wing point See section 2 6 3 6 Image AND Image z Click the Image OR Image button A new Display panel showing Image OR Image in the page tab appears showing the image obtained by the operation 2 6 3 8...

Page 359: ...anel 2 On the Display panel displays and overlaps 2 channel image that to be observed See section 2 4 4 2 for overlapping the image display 3 On the overlapped images click right mouse button to show...

Page 360: ...er point will be colored by gray scale Summary Text Box Summary Dye method of vertical and horizontal axis or channel number Mode Option button selected at Data Selection group box Samples processed T...

Page 361: ...hold line vertical and horizontal red line appears The value can be adjust by dragging those lines Threshold Text Box Threshold values of X and Y axis are displayed on Histogram graph It can also be i...

Page 362: ...hreshold value in Y axis A B A B C D x100 Upper left Upper left portion of histogram graph Samples Total numbers of image pixels contained in upper left portion of Dye name1 threshold A A C x100 of Dy...

Page 363: ...graph The rectangular can be positioned by dragging Annotation mode Group Box When Min Max Bounds option button is selected this dialog will appear Annotation Text Box For details refer to the explan...

Page 364: ...gular Minimum Minimum value of X axis rectangular Maximum Maximum value of X axis rectangular Dye name2 limits Y axis range of rectangular Minimum Minimum value of Y axis rectangular Maximum Maximum v...

Page 365: ...ach peak to draw a polygon When the last peak is clicked the peak clicked at first time will be connected with the last peak Free Region Button Free draw Specify region by dragging on image Releasing...

Page 366: ...ngular and ratio to whole region Samples outside region Total numbers of image pixels outside rectangular and ratio to whole region RGB 2nd pictorial figure Color of rectangular To be displayed in 256...

Page 367: ...data set which is displayed in Display panel Selected range Option Button This button appears when series data set is in use The colocalization is processed on to the all image of the data set Draw Bu...

Page 368: ...ysis Mode Group Box is selected the dialog as shown below appears Fig 2 104 Colocalization Processor Dialog box In case that Analysis Mode is Image Measures Image Measurements Group Box It displays re...

Page 369: ...i F N F 2 2 1 Average brightness of wavelength 2 O Overlap In case that the value to be acquired is defined as r i i i i i i i F F F F r 2 2 2 1 2 1 Overlap Index 1 In case that the value to be acquir...

Page 370: ...APPLIED OPERATIONS Image Processing IV OPERATION INSTRUCTIONS IV 2 2 5 6 Page i i A i i F F m 2 2 2 However A i should be i th pixel that belongs to aggregation A that is greater than threhold value...

Page 371: ...TIME Appending in T direction Appending in T direction Z The image files used in appending are subjected to the following restrictions The sizes of the two image files should be identical The numbers...

Page 372: ...s can be appended in the T direction with the following procedure 1 Open the two image files A and B for appending 2 Display the display panel of the first image A at the front 3 Display the Experimen...

Page 373: ...Process panel click the Experiment List button The Experiments in Memory dialog box appears as shown below Fig 2 106 Experiments in Memory Dialog Box The icon of the image file being displayed at the...

Page 374: ...ion buttons appears under the Append button in the Experiment Editor sub panel make new dim Create an image of new observation mode Select the type in the Append on New Dimension dialog box on Z Appen...

Page 375: ...e Processing IV OPERATION INSTRUCTIONS IV 2 2 6 1 Page 8 A new display panel having Append as the window title is created showing the image after appending For appending more than 3 images at a time s...

Page 376: ...appending several image data set There are the following restrictions when you append image files Two image files must be the same image size Two image files must be the same channel number Two image...

Page 377: ...mage files to be appended Image files can be selected by dragging file name in File area Several image files can assign by using Click and Shift key Click in File area Several image files can also ass...

Page 378: ...n information is appended in case of Z direction appending 6 Select the order for appending image data with Sort Group Box The option buttons work as the followings Date Appends in order of date Alpha...

Page 379: ...e time stamp interval that starts with 0 sec In case of Z in Dimension group box The setting is utilized for the Z step interval that starts with 0 um In case of on An in Dimension group box The setti...

Page 380: ...ams Windows NT Explorer command from the Start menu In case of Windows 2000 select Accessories Windows Explorer command from the Start menu As shown below Windows NT or Windows 2000 Explorer will appe...

Page 381: ...wing is an example for Image extraction from an XYZT image data set 1 Display the image to be extract in Display panel 2 Select Experiment Editor sub panel in Process panel 3 By clicking Crop button E...

Page 382: ...pressing No Display Image slices with a certain time interval and or Z step can be selected by pressing Set after set T Step and or Z Step in Equal Division Image slices with a certain rectangular ar...

Page 383: ...ndidates that are short appear in red Selection of grid candidates that are short is done as follows 1 Verification will be done whether or not the same numbers of Z elements are selected against all...

Page 384: ...l bar 2 Select either Rectangular Circle Polyregion or Free Region to assign clipping area When option other than Rectangular is selected the cut out clipping will be done against rectangle that circu...

Page 385: ...mmand in Programs In case of Windows 2000 select Windows Explorer command in Accessories Then Windows Explore appears 3 Double click FVExtract exe in Fluoview of C Drive 4 Assign folder 1 Folder area...

Page 386: ...ing are possible B A XY XYZ XYT XYZT XZ XZT XT Pt Pt XY AN XY XYZ XYT XY AN XYZ XYT XY AN XYZ XYZ XYZ XYZT XYZT XYT XYT XYT XYZT XYZT XYZT XZ XZT XZT XZT XZT XZT XT XT AN Pt Pt Pt Pt AN XY AN XY AN XY...

Page 387: ...d and double clicking this field displays the Enhanced Profile Plot window When a region is specified the bird s eye view is displayed and double clicking this field displays the Intensity Map window...

Page 388: ...f buttons appears as shown below 4 From the displayed buttons click the Line button Poly Line button or Free Line button 5 Specify the straight line polygonal line or free line on the image in the Dis...

Page 389: ...sity profile cannot be displayed when the handles are not displayed The checked line can be moved deleted or changed of size or color This is possible with the same method as entering comment in the i...

Page 390: ...the bottom left to the right of the magnified graph When the mouse pointer is placed on a graph line while the Ctrl or Alt key is held depressed the coordinates can be displayed The displayed data can...

Page 391: ...the multiple channels are analyzed simultaneously or only one channel is analyzed To select the target channel s use the Display channel switch buttons Only the channel s being displayed will be anal...

Page 392: ...nnect the last clicked point to the first clicked point z To specify a free region On the image specify a region by dragging Then release the mouse button to complete dragging The point where the drag...

Page 393: ...as the area Area horizontal and vertical lengths X Y Z or T and perimeter length Perimeter of the region and the statistic result of each channel such as the total Total average Average and standard...

Page 394: ...compatible format Copy button Copies the plotted image in the clipboard Close button Quits the Enhanced Profile Plot window and returns to the Analyze panel Tilt scale Sets the angle in the vertical d...

Page 395: ...gram Plot windows appears as shown below Fig 2 114 Enhanced Histogram Plot Window Line Specification When a desired area is specified by dragging the left button of the mouse on the graph the specifie...

Page 396: ...e histogram is displayed in the Region Histogram box in the Single sub panel The operation method is identical to displaying the intensity profile on a line See section 2 7 1 2 Intensity Values on a P...

Page 397: ...the graph can be scrolled The magnification or scrolling of the graph can be canceled by dragging the left button of the mouse from the bottom left to the right of the magnified graph When the mouse...

Page 398: ...ethod for measuring the perimeter of a region is identical to that for displaying the bird s eye view of a region See section 2 7 1 2 Intensity Values on a Planar Region Bird s Eye View The measuremen...

Page 399: ...lected the Single Slice Integration group box appears as shown below Select the direction of interest X or Y using the option buttons When the selected image was acquired in the XY observation mode th...

Page 400: ...ross section Z time lapse T to be subjected to the operation can be selected using the XYZ series and XYT series buttons above the image 4 When the image was acquired in the multi channel mode select...

Page 401: ...see section 2 7 1 2 Intensity Values on a Planar Region Bird s Eye View A region is displayed on the image together with handles on the perimeter The region is selected as the target of mean value co...

Page 402: ...simultaneously and display their operation results together First specify the regions by repeating steps 7 and 8 above for each Use different colors for the regions After having set the regions click...

Page 403: ...e of the specified regions will be displayed graphically in the Mean Intensity box The colors of the chart lines corresponding to the colors assigned to the regions When the image was acquired in the...

Page 404: ...the bottom left to the right of the magnified graph When the mouse pointer is placed on a graph line while the Ctrl or Alt key is held depressed the coordinates can be displayed The displayed data can...

Page 405: ...measured and displayed graphically The operation results are displayed graphically in the Integrated Intensity box in the Series sub panel The operation method is completely identical to that for obta...

Page 406: ...aph When the mouse pointer is placed on a graph line while the Ctrl or Alt key is held depressed the coordinates can be displayed The displayed data can be applied to other applications See section 2...

Page 407: ...he Display panel of the XYZ multiple sections image 2 The following button is displayed at the top of the Display panel Usually only the XYZ series button is displayed When it is clicked a list of but...

Page 408: ...n Image from a Different Viewpoint IV OPERATION INSTRUCTIONS IV 2 2 9 4 Page Image No 0 Image No 1 Image No 2 Image No 3 Fig 2 120 Four Images Used in Building Extended Focus Image Fig 2 121 Panel Sho...

Page 409: ...be set as the end image by using the Display buttons at the top of the Display panel 4 Click the Set end position button at the top of the Display panel The Set start position and Set end position bu...

Page 410: ...y the Display panel of the XYZ multiple sections image 2 Click the Extended Focus View option button in the Rendered View group box 3 Click the Begin Visualize button to start the image building When...

Page 411: ...APPLIED OPERATIONS Building an Image from a Different Viewpoint IV OPERATION INSTRUCTIONS IV 2 2 9 7 Page Fig 2 123 Panel Showing Extended Focus Image...

Page 412: ...e 2 Click the Extended Focus View option button in the Rendered View group box 3 Click the Begin Visualize button to start the image building When it completes the built image is displayed in the Exte...

Page 413: ...or YZ direction on the XY direction image displayed left above Make Multi Plane View button Image display in XY direction The line image to be displayed in XZ or YZ direction can be moved by dragging...

Page 414: ...color red green eyeglasses Sum option button Builds the image by adding the intensity values Rendered View group box Extended Focus View option button Checking this option button changes the check bo...

Page 415: ...ossible to provide the image with a perspective by darkening the far objects and brightening near objects Stereo Factor text box Sets the deviation between the left and right eyes when building a pair...

Page 416: ...e by frame display or to stop successive display Holding the button displays image slices in successive display and clicking it displays image slices by advancing frame by frame Display button Press t...

Page 417: ...e Successive Display Speed The buttons on the top of the Display panel can be used to vary the speed of successive display of multiple image slices 1 Display the Display panel of the image to be subje...

Page 418: ...osition Entering the value directly in the text box can also move the display position 2 9 2 Animation Images composed of multiple image slices acquired by varying the multiple sections XYZ observatio...

Page 419: ...ews drop down list 6 Click the Initial view option button 7 Set the angle at which the rotation should start using the Initial Rotation Angle scale 8 Click the Sequence views option button 9 Set the a...

Page 420: ...utton Click the Cancel Visualize button if you want to cancel the building Fig 2 132 Panel Showing the Built Image 12 Click and hold the Display button at the top of the 3D Animation panel The images...

Page 421: ...e button to start building the images When the building completes the built images are displayed in the 3D Animation panel The status bar shows the progress of building processing During the building...

Page 422: ...APPLIED OPERATIONS Viewing 3D Image IV OPERATION INSTRUCTIONS IV 2 3 0 8 Page Fig 2 133 Panel Showing Stereo 3D Images...

Page 423: ...sualize button to start building the images When the building completes the built image is displayed in the 3D Animation panel The status bar shows the progress of building processing During the build...

Page 424: ...APPLIED OPERATIONS Viewing 3D Image IV OPERATION INSTRUCTIONS IV 2 3 1 0 Page Fig 2 134 Panel Showing 3D Image to be Viewed Through Color Eyeglasses...

Page 425: ...Images Successively The change over time can also be displayed as animation With images composed of multiple slices such as time lapse images the buttons as shown below are displayed at the top of th...

Page 426: ...ays image slices in successive display and clicking it displays image slices by advancing frame by frame Display button Press to start display of n slices of image from the end position to the start p...

Page 427: ...is After it display the Enhanced Profile Plot Intensity Map Enhanced Histogram Plot Average Intensity Trace or Integrated Intensity Trace For the operation method see section 2 7 Image Analysis Fig 2...

Page 428: ...ialog Box 3 Exit from FLUOVIEW or display the Start menu by pressing the Ctrl Esc keys 4 Select Programs and issue the Microsoft Excel command 5 From the File menu of Excel select the Open command and...

Page 429: ...Tab check box in the Delimiters group box then select none from the Text Qualifier drop down list Fig 2 138 Dialog Box When the File is Opened by Excel 2 3 8 Click the Next button When the dialog as s...

Page 430: ...cation handling images such as Paint The following description takes Paint as example 1 Display the Analyze panel and executes analysis After it display the Enhanced Profile Plot Intensity Map Enhance...

Page 431: ...Paint 2 11 3 Transferring Image Data to Another Application microVoxel Paint etc To transfer image data to another application the image data should be saved in a file and the file should be transferr...

Page 432: ...appears as shown below 2 12 1 Writing Characters in Image This facility is used to enter the title acquisition parameters and or notes in an image Some labels are provided in advance Characters can be...

Page 433: ...zoom z t animation channel stereo bounds Desired characters can also be entered 1 Click the character in the drop down list of the dialog box 2 Delete the character by pressing the Delete or Back Spa...

Page 434: ...ns of the Active Overlays dialog box 2 12 2 Displaying the Image Intensity The intensity of any pixel of an image can be displayed without using the Analyze panel 1 Display the Display panel of the im...

Page 435: ...the Text button The dialog box as shown below appears Fig 2 144 Active Overlays Dialog Box 3 In the drop down list inside the dialog box select x x hotspot value units to display the X coordinate pos...

Page 436: ...tion To draw a rectangle On the image drag the mouse pointer along the diagonal line of the desired rectangle from the top left corner to the bottom right corner To specify a circle or ellipse On the...

Page 437: ...2 11 8 Changing the Comment Size for the operation procedure To display a correct scale it is required that the image has been acquired while the objective magnification setting in the software match...

Page 438: ...he Text button Perform the following operation before starting to draw the scale 1 Click the Text button in the list of buttons 2 When the Annotation dialog box is displayed click its Font button The...

Page 439: ...the displayed list of buttons 3 Drag the mouse pointer from the start point to the end point of the desired arrow 4 To change the arrow size see section 2 11 8 Changing the Comment Size The direction...

Page 440: ...ging the mouse pointer along the desired position in the image 4 To change the color bar size see section 2 11 8 Changing the Comment Size The labels of the color bars can be switched to display or hi...

Page 441: ...one of the comments and click the moue while holding the Shift key to select the second comment and after After making all of the comments to be deleted active i e handled displayed around them click...

Page 442: ...isplayed In this case use the method of displaying the pop up menu 2 12 10 Changing the Comment Size 1 Click the mouse on the comment to be resized to make it active i e handles displayed around it 2...

Page 443: ...A pop up menu as shown in Fig 2 145 appears 3 Select Properties from the menu The Properties dialog box as shown below appears Display the Color panel at the front Fig 2 146 Properties Dialog Box 4 Se...

Page 444: ...ont to make the comment active i e handles displayed around it 2 Click the right button of the mouse A pop up menu as shown in Fig 2 145 appears 3 Select Properties from the menu The Properties dialog...

Page 445: ...as shown below appears Fig 2 148 Print Dialog Box 3 Select the connected printer name from the Name drop down list 4 If it is required to set the detailed data of the printer click the Properties butt...

Page 446: ...ne Point The Print dialog box can also be displayed by mouse operation on the image 1 Display the image to be output at the printer at the front of the Display panel and click the right button of the...

Page 447: ...Press to start display of n slices of image from the start position to the end position sequentially successive display or one frame by frame display or to stop successive display Holding the button d...

Page 448: ...automatically 2 14 2 Merging Image Channels For instance when a specimen is imaged with two scans for 2 channel fluorescence observation and 1 channel transmitted light observation the transmitted lig...

Page 449: ...on procedure When the images in both image files are composed of multiple image slices the overlaid image will have the same number of image slices as the file with the fewer image slices Example 1 XY...

Page 450: ...the Experiments in Memory dialog box select the file name of the first image and drag it into the frame at the top The icon of the first image is displayed in the frame at the top left The mouse poin...

Page 451: ...1 Navy blue Ch2 Light blue Ch3 Green Ch4 Yellow green Ch5 Orange Ch6 Red When the mouse pointer is approached to the icon side end of a line connected to Merger the color of the line end turns into ye...

Page 452: ...OPERATION INSTRUCTIONS IV 2 3 3 8 Page 7 Click the Merge button in the Experiments in Memory dialog box A new Display panel showing Merge in the page tab appears and the images including newly merged...

Page 453: ...the image to extract channels in advance When the image has been saved in more than one image file it is possible to use only some of the slices by setting a slice range See section 2 14 1 Setting th...

Page 454: ...ick the Extract channels button The Experiments in Memory dialog box appears as shown below Fig 2 153 Experiment Editor Group Box The frame at the top left of the Experiments in Memory dialog box show...

Page 455: ...t in the Experiments in Memory dialog box are connected to Extract by lines The lines of channels are colored as shown below Ch1 Navy blue Ch2 Light blue Ch3 Green Ch4 Yellow green Ch5 Orange Ch6 Red...

Page 456: ...eselect the unnecessary channels as described in the TIP on the previous page so that only the necessary channels are selected 7 Click the Extract button in the Experiments in Memory dialog box A new...

Page 457: ...chart display method to be changed 2 15 1 Chart Panel z Series panel This panel is used to change the type title or other settings of the displayed chart as well as to add or delete charts List of di...

Page 458: ...art or sets the orientation and margins of the print paper Save the chart in the format of an image file or in the clipboard Set the margins from the edges of the area in which the chart is displayed...

Page 459: ...y selecting the optimum scale automatically Set the number of steps of the values displayed in the chart axis labels Enable or disable the inverted chart display Set the title of the coordinate axis S...

Page 460: ...along the label Enable or disable the display of coordinate axis scales in the intermediate position between labels Set the length of the scale line from the coordinate axis to the label Enable or di...

Page 461: ...to the start point Set the coordinate axis display range with reference to the end point Switch the title display position between the header and footer Enable or disable the title display Switch the...

Page 462: ...the margin between the chart display area and legend display area Set the color and width of the shadow area of the shadowed display of legend area Enables or disables the inversion of the display or...

Page 463: ...ay Changes color of Left Bottom and Back wall Background button Sets the background color Border button Sets whether or not the background frame is displayed and sets its color width and type Pattern...

Page 464: ...e direction Enables or disables the zooming display of character strings in the chart such as label characters together with the chart display Change the zoom ratio Rotates the chart in the vertical d...

Page 465: ...panel when the Point 3D chart is selected Sets whether or not the outer frame of the chart is displayed and sets the color width and type of the frame Enables or disables the shadowed 3D display Enabl...

Page 466: ...inflection points Sets the width of inflection points Sets the height of inflection points Sets the type of inflection points Sets the inflection point background color Resets the background color to...

Page 467: ...ondition Enables or disables the item display in the chart Sets the background color of the item display area Enables or disables the transparent background display Sets the item character font Sets t...

Page 468: ...ing comment setting can be controlled by clicking the right button of the mouse without selecting specific page tabs or buttons zPop up menu of function panel When the right button of mouse is clicked...

Page 469: ...right button of mouse is clicked on the page tab of a Display panel a pop up menu appears to allow selection of the Display panel to be displayed at the front Fig 2 155 Display Panel Showing Pop up Me...

Page 470: ...ht button of mouse is clicked on an comment in image a pop up menu appears to allow editing or deleting the specified comment Fig 2 156 Image Comment with Pop up Menu Moves the comment Resizes the com...

Page 471: ...image save LUT setting number of image divisions comment editing Fig 2 157 Display Panel Showing Pop up Menu Displays the image full screen Outputs the image at the printer Saves the image as a serie...

Page 472: ...image file in the File I O Tile or Process panel a pop up menu appears This makes it possible to select the file to be opened when performing tile display inter image operations image channel merger e...

Page 473: ...g the LUT Look Up Table Exit button Exits from FLUOVIEW Annotate button Allows the analysis area to be specified or comment to be drawn in the image Print button Outputs the image at the printer Exper...

Page 474: ...olding the button displays image slices in successive display and clicking it displays image slices by advancing frame by frame Set start position button When successive display or frame by frame disp...

Page 475: ...ation m Enables and makes variable the LED slider on the left Enables and makes variable the LED slider on the right p Decreases the value of an enabled LED slider Fine adjustment Shift p Decreases th...

Page 476: ...Operation Page Up Moves the Z stage up Fine adjustment Shift Page Up Moves the Z stage up Coarse adjustment Page Down Moves the Z stage down Fine adjustment Shift Page Down Moves the Z stage down Coa...

Page 477: ...natively switches over between the previously assigned LUT and Hi Lo LUT on the channel whose PMT adjustment window PMT Gain and Offset is opening on the Acquire panel during the acquisition period Ct...

Page 478: ...ub panels Key Target Operation Alt M Selects the Math sub panel Alt I Selects the Filter sub panel Alt H Selects the Histogram sub panel Alt K Selects the Mask sub panel It is an option panel with TIE...

Page 479: ...mand button A figure in the shape of button in the window Clicking a command button with the mouse allows the function indicated on the button to be executed Confocal Signifies the possibility of obta...

Page 480: ...icon of the window I Icon An icon is a small figure with characters below it The icon indicates the status in which the window is closed or minimized Iconize This refers to turn a window into an icon...

Page 481: ...usually provides a better effect than increasing PMT Voltage above 800 V R Resolution Number of dots composing the image on a screen or printer When the number of dots is increased i e when the resol...

Page 482: ...indicates the position where the input character is inserted Title bar The horizontal bar at the top of the window that shows the title of the window or dialog box Toolbar The toolbar provides freque...

Page 483: ...d only by a person who have the authority of Administrator 1 Click the start button at the bottom of the Windows screen 2 When the start menu is displayed select commands Programs Administrative Tools...

Page 484: ...of the Create partition of size edit box in the Create Primary Partition dialog box to Maximum size and click the OK button 6 From the Partition menu of the Disk Administrator window select the Commit...

Page 485: ...the Format dialog box Then select FAT from the File System drop down list and select FAT and select Default Allocation Unit Size from the Allocation Unit Size drop down list 12 Click the Start button...

Page 486: ......

Page 487: ...3 When the dialog box as shown below appears click the Delimited option button in the Original Data Type group box then select Windows ANSI from the File Origin drop down list Fig Appendix E 1 Dialog...

Page 488: ...Page 5 Click the Next button When the dialog box as shown below appears click the General option button in the Columns Data Format group box then click the Finish button Fig Appendix E 3 Dialog Box Di...

Page 489: ...s to be inserted The dialog box as shown below appears Fig Appendix E 4 Dialog Box of Chart Wizard 1 5 9 Click the Next button The dialog box as shown below appears Fig Appendix E 5 Dialog Box of Char...

Page 490: ...rd 4 5 12 Click the Columns option button under Data Series in 13 Set the data column number that you want to use as the X axis label position in the Use First Column s text box 14 Click the Next butt...

Page 491: ...se is clicked on the chart black handles will appear around the chart The following operations are available in this condition z Moving the chart Place the mouse pointer on the chart and drag it z Res...

Page 492: ...k handles 2 Double click The dialog box for setting the selected format appears Set the desired format and click the OK button Refer to the Excel manuals for details To return to the original conditio...

Page 493: ...Use the following procedure to register yourself as a user Log in the system with Administrator Register yourself as a new user 1 From the Start menu select Programs Administrator User Manager 2 The U...

Page 494: ...r in the Full Name text box 7 Enter the description on the user in the Description text box 8 Enter a password with up to 14 characters in the Password and Confirm Password text boxes 9 Click the User...

Page 495: ...Register yourself as a new user 1 Double click the FLUOVIEW Setup icon on the desktop Then the dialog box as shown below appears 2 Click the Users icon in Hardware Settings to display the FLUOVIEW Us...

Page 496: ...and click the OK button The newly set user name is added in the list box 6 To register other users repeat steps 3 to 5 for each one 7 Click the Save New Setup button or Quit w o Saving button to close...

Page 497: ...me appears Enter the user name to log in the FV300 1 Double click the FLUOVIEW icon on the desktop The dialog box for entering the user name appears as shown below 2 Enter the user name in the User Na...

Page 498: ...ge Appendix G 3 Deleting a User 1 Double click the FLUOVIEW icon on the desktop The dialog box for entering the user name appears as shown below 2 Enter Administrator in the User Name text box and cli...

Page 499: ...tton The Administrator cannot be deleted from the list 6 The Delete a user dialog box appears to ask if you really want to delete the user NOTE Add a user and Log in button Used to add a new user Rese...

Page 500: ...Page 7 Click the Yes button if you want to delete the user or the No button if you do not The deleted user name disappears from the list The user is deleted at the moment the OK button is clicked 8 C...

Page 501: ...mage can be changed or a desired folder can be specified directly when loading a saved image Change the default folder only when required There is no need of change if folder C FLUOVIEW IMAGES is all...

Page 502: ...ault Folder for File I O Panel IV OPERATION INSTRUCTIONS IV H 2 Page 1 Click the Start button to open the Start menu Then select commands Settings Control Panel System The System Properties dialog box...

Page 503: ...lect the Environment sub panel 3 Select FLUOVIEWIMAGES under Variable in the User Variables for Administrator list box Administrator of User Variables for Administrator is variable depending on the us...

Page 504: ...drive and folder names using backslashes The figure shows an example in which folder IMAGE in drive C is entered 5 Click the Set button Value text box Enter the path name of the default folder Change...

Page 505: ...e changed from the next time you start FLUOVIEW z When the default folder is changed to C IMAGES To reset the default folder to the factory setup enter C FLUOVIEW IMAGES in the Value text box Do not c...

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Page 507: ...tion with respect to the top left corner of the screen is displayed Syntax x hotspot raw calibrated value units Arguments inside can be omitted If raw calibrated is omitted the same setting as when ca...

Page 508: ...ed will be applied Setup Procedure 1 Enter y as the first character inside 2 Add the following characters to set the display method hotspot Display measurement points with markings raw Display data in...

Page 509: ...de can be omitted If raw calibrated is omitted the same setting as when calibrated is specified will be applied 2 T Position With time related images such as XYZ images the T position with respect to...

Page 510: ...ecified will be applied Setup Procedure 1 Enter intensity as the first character inside 2 Add the following characters to set the display method hotspot Display measurement points with markings raw Di...

Page 511: ...s are overlapped the displayed channel numbers are connected by the markings Syntax Channel Appendix I 3 2 Objective Power The objective power used in image capturing can be displayed Syntax Objective...

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Page 513: ...tment knob are shipped from the factory the following functions have been allocated to their control buttons These function allocations are valid only when the FLUOVIEW software is started Appendix J...

Page 514: ...ction 1 Stage escape return switching 2 Focus fine coarse adjustment switching 3 Z position upper limit setting in Z observation 4 Z position lower limit setting in Z observation 5 Excitation DM posit...

Page 515: ...Function 1 Focus fine coarse adjustment switching 2 Excitation DM position up 3 Excitation DM position down 4 BI LSM light path switching 5 Objective revolving nosepiece position up 6 Objective revolv...

Page 516: ...limit setting in Z observation 4 Z position lower limit setting in Z observation 5 Excitation DM position up 6 Excitation DM position down 9 Focus fine coarse adjustment switching 2 IX Fig Appendix J...

Page 517: ...describes the user maintenance procedures of the FLUOVIEW FV300 system Please read this volume so that you can use the system for an extended period of time On This Volume V V M MA AI IN NT TE EN NA...

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Page 519: ...ion 1 6 1 3 1 Overall Setting of FLUOVIEW 1 6 1 3 2 Setting the Microscope Control Panel 1 14 1 1 Adding the dyeing method 1 21 1 1 1 Associating a Detection Channel and Filter to the Dyeing Method 1...

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Page 521: ...To set up the FLUOVIEW software it is necessary that the computer in use already has Microsoft Windows NT Version 4 0 English version or Microsoft Windows 2000 English version installed in its hard di...

Page 522: ...tware V MAINTENANCE V 1 2 Page 4 Continuously deletion of software is started For deletion carry out according to a wizard 5 Continuously installation of the software of a new version is started Refer...

Page 523: ...CD ROM in the CD ROM drive The dialog box as shown below will appear If the dialog box doesn t appear run the Install exe file that is present in the root directory of the CD ROM 2 Select the Open Rea...

Page 524: ...tware Setup New Setup of the Software V MAINTENANCE V 1 4 Page 5 When the Choose Destination Location dialog box appears confirm the setup destination drive name and directory and select the Next butt...

Page 525: ...the Software V MAINTENANCE V 1 5 Page 6 When the setup has completed the Setup Complete dialog box appears Select the Yes I want to restart my computer now option button and press the Finish button Th...

Page 526: ...and click the OK button to log into FLUOVIEW FV300 For details see Appendix G 2 Logging into the FV300 3 The dialog as shown below appears for use in saving the system configuration First select the...

Page 527: ...Start Z and Stop Z text boxes in the Z Stage sub panel are set according to the value set there For example When 1 0 mm is set as the maximum range for Z series the minimum value for Z starting become...

Page 528: ...shown below appears select the Delete button To change the details objective name pinhole No magnification of an objective Double click the objective for which you want to change the detailed informa...

Page 529: ...with FLUOVIEW The setting procedure is similar to the objective setting procedure Objectives panel 7 Click the Coloring Tables icon in User Settings to display the Coloring Tables panel in the front p...

Page 530: ...ult Image Zoom in Single View group box Select the initial status of display magnification when a single image is to be displayed in a single Display panel Default Image Zoom in Multiple View group bo...

Page 531: ...ning when setting fast scan mode Delay Timing for TD Unit group box This data should usually be left in the default values and does not have to be changed Delay shutter changes sec text box Enter the...

Page 532: ...x Set the laser intensities when starting up the FLUOVIEW application User default intensities option button Select to use the default intensity set in the Default Laser Intensity scale above Use most...

Page 533: ...3 Page 12 The Registration panel have been set at the factory and do not need to be changed here 13 After completing the setup select the Save New Setup button on the bottom left of the panel Selecti...

Page 534: ...ope Setup button Displays the Microscope Setup window Mirror Unit group box Sets up the name and color of the cube turret Nosepiece group box Sets up the name color of the objective and sets the magni...

Page 535: ...nser Turret group box Sets up the name and color of the universal condenser Filter Turret group box Sets up the name and color of the filter turret for reflected light observation Filter Turret group...

Page 536: ...me Editing in the Nosepiece group box x Setting the Name x Setting the Objective Magnification x Setting the condenser worked with x Setting the magnification of the objective x Setting the N A of the...

Page 537: ...Nothing disables the turret itself 1 3 2 2 Setting the Name 1 Editing the name of the buttons in the Mirror Unit Condenser Turret and Filter Turret group boxes 1 In the Mirror Unit Condenser Turret or...

Page 538: ...shown below appears 2 Select Color The Color dialog box as shown below appears 3 Select the color which you want to set in the color palette and click the OK button TIP Name of the Objective drop dow...

Page 539: ...pecify the color of the button of the cube the objective and the condenser engaged into the light path Right click the mouse on the area outside of the buttons in the Mirror Unit Nosepiece Condenser T...

Page 540: ...window Or click the Quit w o Saving button to cancel the setup and close the window Then the dialog box as shown below appears If you want to save click the Yes button if you do not want to save click...

Page 541: ...laser type excitation wavelength and emission wavelength Add the dyeing method using the Fluorescent Dyes Colors panel in the Fluoview Setup dialog box 1 Double click the FLUOVIEW Setup icon on the d...

Page 542: ...ld be set in advance When HeCd is not displayed check the HeCd check box in the Laser Equipment panel 7 Enter the value of the excitation wavelength into the WaveLength 1 text box in the Excitation Wa...

Page 543: ...button to close the Edit Dye dialog box 10 Select the Save New Setup button to close the FLUOVIEW Setup dialog box Some dyeing method does not appear in Available Dyes list box in Dyes subpanel It ha...

Page 544: ...and filter can be associated to a dyeing method using the Optical System Configuration Setup dialog box 1 Display FLUOVIEW Setup dialog See section 1 3 1 of this manual to display FLUOVIEW Setup dialo...

Page 545: ...g method select the required methods while holding the Ctrl key TD Unit group box Displayed when the transmitted light unit is installed The channel No is displayed together with Transmitted Dyes drop...

Page 546: ...n DM drop down list When it is not required to specify excitation dichroic mirror select the blank In this example select DM488 515 6 To change the beam splitter use the Beam Splitter drop down list W...

Page 547: ...Adding the dyeing method V MAINTENANCE V 1 2 7 Page the blank In this example select CFP for channel 1 9 After completing the above click the Save button to close the Optical System Configuration Set...

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Page 549: ...ing these parts and for adjusting the light axis after replacement 2 1 1 Replacing the Barrier Filters Slider To replace the BARRIER FILTERS slider loosen the filter slider cover screws 10 and remove...

Page 550: ...s or the dangerous laser come out For details refer to the instruction manual of your microscope The other hand in the United States of America this product is classified as a CLASS a laser product ac...

Page 551: ...be performed by the user for the purpose of assuring the intended performance of the product It does not include operation or service The definitions of the term maintenance are quoted below for refe...

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Page 553: ...ssible troubles In case of trouble please read volume before calling for service If the normal operation cannot still be restored please contact your local Olympus representative On This Volume V VI I...

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Page 555: ...position reproduction of the Z motor is poor 1 4 12 The Acquire panel cannot be displayed 1 4 13 The scale of the Z Stage sub panel in the Acquire panel cannot be moved 1 4 14 The fine movement knob o...

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Page 557: ...2 4 OPERATION INSTRUCTIONS An excitation tube or analyzer for visual observation is engaged in the microscope Disengage the excitation tube and analyzer from the light path 1 2 4 OPERATION INSTRUCTION...

Page 558: ...ox in the Acquire panel 1 1 1 2 2 1 3 3 OPERATION INSTRUCTIONS 2 Transmitted image cannot be observed The transmittance detection channel is not selected In the Acquire panel check the check box of th...

Page 559: ...uorescent glass is not used Use non fluorescent glass The cover glass thickness is not optimum Use a cover glass with a thickness of 0 17 mm The specimen is over stained Stain it properly again or inc...

Page 560: ...ck the Locked check box in the Z Stage sub panel 2 2 1 3 7 OPERATION INSTRUCTIONS 14 The fine movement knob of microscope cannot be turned or is not smooth The Z stage Z revolving nosepiece motor is e...

Page 561: ...the mouse The software is malfunctioning Press Ctrl Alt Delete on the keyboard and exit from FLUOVIEW by following the displayed messages Then exit from windows and reboot the computer 1 2 1 1 2 2 OP...

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Page 564: ...Drive Melville NY 11747 3157 U S A OLYMPUSSINGAPOREPTELTD 491B River Valley Road 12 01 04 Valley Point Office Tower Singapore 248373 OLYMPUSOPTICALCO U K LTD 2 8 Honduras Street London EC1Y OTX United...