Olympus FV1000 User Manual

User’s Manual
A
FLUOVIEW
FV1000
CONFOCAL LASER SCANNING
BIOLOGICAL MICROSCOPE
[OPERATION] FV10-SW Ver 5.0c
Petition
This user’s manual is for the software to be run on Olympus FLUOVIEW FV1000 Confocal Laser Scanning Biological Microscope. To ensure safety, obtain optimum performance and familiarize yourself fully with this product, we recommend that you study this manual thoroughly before operation. This user’s manual is composed of two volumes including “OPERATION INSTRUCTIONS” and “MAINTENANCE”. Together with this manual, please also read the “SAFETY GUIDE”, “HARDWARE GUIDE” of User’s manual FLUOVIEW FV1000 and the instruction manual of the microscope in order to understand overall operation methods. To ensure the safety operation of laser system, we recommend you to study the manual of each laser and the light source equipment besides this manual. Retain this manual in an easily accessible place near a system for future reference.
X7274

CAUTION

CAUTION
1. Reproduction, copying or duplication of a part or all of this software and manual is prohibited.
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 Corporation.
Other brand names and product names are trademarks or registered trademarks of their respective owners.
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FLUOVIEW MANUAL CONFIGURATION

FLUOVIEW MANUAL CONFIGURATION
The FLUOVIEW system uses two manuals including this “User’s Manual” and the on-screen
manual built into the software (“Online Help”).
The User’s Manual is composed of the five following volumes and subject matter:
OPERATION INSTRUCTIONS
Describes the operation procedures of the FLUOVIEW system, for example, methods
for image acquisition and various image processing.
1 Getting Started FLUOVIEW................................................................................ 1-1
2 APPLIED OPERATIONS.................................................................................... 2-1
Appendix A List of Hot Keys.................................................................................. A-1
Appendix B Glossary............................................................................................. B-1
Appendix C USER REGISTRATION OF FV1000 ................................................. C-1
Appendix D Change of Default Folder for [File I/O] Panel..................................... D-1
Appendix E List of Functions in the [Active Overlays] Dialog Box......................... E-1
Appendix F Hand Switch and Microscope Frame Function Allocation .................. F-1
MAINTENANCE
Describes maintenance of the FLUOVIEW system.
1 Software Setup................................................................................................... 1-1
2 Maintenance of Major System Units................................................................... 2-1
3 Setting the Confocal Aperture ............................................................................3-1
TROUBLESHOOTING
Describes countermeasures in case trouble occurs.
1 TROUBLESHOOTING GUIDE ........................................................................... 1-1
For Online Help, please see “1-3 Online Help” in “OPERATION INSTRUCTIONS” of this
manual.
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NOTATIONS IN THIS MANUAL

This manual complies with the following notations.
Notation of Caution, Notes and Tips
Notation Description
NOTATIONS IN THIS MANUAL
Caution to prevent injuries to the user or damage to the
product (including surrounding objects).
NOTE
TIP
Note for the user.
Hint or one-point advice for user reference.
Notation of panel, Command Buttons and Dialog Boxes
Notation Description
[Acquire] panel The name of a panel, dialog box, list box or check box is
enclosed inside square brackets ([ ]).
<OK> button
<Open File> button
The name of a button in a panel or dialog box is enclosed
inside angled brackets (< >).
Notation of Mouse Operations
Notation Description
clicking Action of pressing, then immediately releasing the mouse
button.
double-clicking Action of clicking the mouse button twice in quick succession.
dragging Action of moving the mouse while holding down the mouse
button, then releasing the mouse button at the desired
destination.
(Note) In this manual, clicking, double-clicking and dragging involves pressing the left button
of the mouse, unless otherwise specified.
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3
NOTATIONS IN THIS MANUAL
Notation of key operations
Notation Description
Enter
The name of a key is enclosed inside .
Alt
+
Direction keys
F1
The positive sign (+) expresses the combination of more than one key operation.
For example,
key while holding the
F1
Generic names given to the
keys.
Alt
+
refers to pressing the
F1
key down.
Alt
,
,
and
Notation of system-specific terms
Notation Description
XY observation
(Other observations)
Note that some of the panels and dialog boxes shown in this manual are not the precise
reproductions of the originals. Some windows are resized to facilitate the reading and some
grayed-out characters are printed in readable characters.
Refers to observation with XY scanning.
(The same principle also applies to other observations such
as XZ, Xt, XYZ, XYt and XYZt.)
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Software Functional Configuration

This software uses panel-type windows.
Usually, it is required to “select a menu then select the command to be executed” in order to
execute a function provided by software. With the panel system, a software function can be
executed easily by “selecting the panel page tab of the function to be executed”, just like
when using a system notebook or file folder.

Function Window and Image Window

The FLUOVIEW software is organized by two kinds of windows, the function windows and
the image .window.
The function windows include the [Acquire], [File I/O], [Tile], [Process], [Analyze] and
[Visualize] panels.
The image .window shows either the [Live] panel or the panel image loaded from a file
([(filename)] window).
The function window
Software Functional Configuration
NOTE
The image window
In this manual, the function windows are referred to simply using their
page tabs.
Namely, the [File I/O] panel of function windows is referred to simply as the
[File I/O] panel.
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5
Software Functional Configuration
A
A

Panel Structure of the Software

This software cannot show the all function panels at a glance.Please use the following list of
the panels for reference in scrolling.
Acquire
Settings
Z Stage
Time Series
Dyes
Lasers Sets the intensity of laser and other option for image acquisition.
File I/O
Tile
Process
Math
Filters
Histogram
Experiment Editor
Analyze
Single
Sets up the image acquisition and executes actual acquisition.
Sets the zooming ratio and observation mode for image acquisition. Sets the microscope light path for image acquisition.
Sets the Z-direction scanning range for image acquisition.
Sets the interval period for image acquisition.
Sets the fluorescence dye method for image acquisition and reagent for each channel.
Saves, loads and deletes images.
Changes the image display method.
Processes the acquired images.
Performs mathematical and logical operations between images.
Filters images.
Changes the image contrast.
ppends two images, adds and extracts the channel.
nalyzes image data.
Obtains the intensity values, intensity distribution, length, area and average intensity in images.
Series
Visualize
Orientation
Other Options
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Obtains the change in the sum of intensity values in images.
Constructs an image from a different viewpoint or displays a 3D image.
Sets the image rotation angle, direction and number.
Sets the various 3D Rendering.

Icons Executed 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 an icon (image file or observation method), dragging it to the desired position and
dropping it there”.
Software Functional Configuration
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7
Software Functional Configuration
Identification of Images Depending on the Observation Methods
Image Icon Significance
XZ observation
XZ observation, 2-channel mode
Xt observation
Xt observation, 2-channel mode
XZT observation
XZT observation, 2-channel
mode
XY observation
XY observation, 2-channel mode
XYt observation
XYt observation, 2-channel mode
On many occasions, FLUOVIEW displays image
icons to allow identification of the observation
method used when each image is acquired. (See
table on the left.)
When the [File I/O], [Tile], [Process], [Analyze] or
[Visualize] panel is selected, the icon of the image
selected in the image window is displayed in a
frame at the top of the function panel. The image
icons are also displayed 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.
TIP
In all observation modes, the icons for 3
XYZ observation
XYZ observation, 2-channel mode
XYZt observation
XYZt observation, 2-channel
mode
Point Scan
Animation image
Stereo 3D image: Image to be viewed with color eyeglasses.
3 or more channels
or more channels are identical.
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O
O
P
P
E
E
N
IIN
R
R
S
A
A
S
TII
T
T
T
R
R
O
O
U
U
N
N
C
C
TII
T
O
O
N
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S
S
On This Volume
This volume describes the operating procedures of the
FLUOVIEW FV1000 system.
“Getting Started FLUOVIEW” contains information on the basic
operation flow until acquisition of XY images.
“APPLIED OPERATIONS” provides detailed operating
procedures of the system.
Please read this volume so that you can understand the system
before use.

CONTENTS

1 Getting Started FLUOVIEW 1-1
1-1 Basic Operations .............................................................................1-1
1-1-1 Microscope .................................................................................................... 1-1
1-1-2 General Mouse Operation Procedures.......................................................... 1-7
1-1-3 Names of Major Panel and Window Controls and Their Functions............... 1-8
1-2 Outline of LSM Observation Procedures ......................................1-9
1-2-1 Turning Power On ....................................................................................... 1-11
1-2-2 Focusing on the Specimen.......................................................................... 1-12
1-2-2-1 Combination with BX .......................................................................................1-12
1-2-2-2 Combination with IX81 FVF .............................................................................1-14
1-2-3 Setting the LSM Light Path.......................................................................... 1-16
1-2-3-1 Combination with Upright Microscope (BX).....................................................1-16
1-2-3-2 Combination with Inverted Microscope (IX81 FVF) .........................................1-18
1-2-4 Selecting the Dyeing Method ...................................................................... 1-20
1-2-5 Selecting the Filters..................................................................................... 1-23
1-2-6 Setting the ND Filters ..................................................................................1-26
1-2-7 Setting the Observation Condition............................................................... 1-27
1-2-7-1 Setting the Objective Magnification .................................................................1-27
1-2-7-2 Setting the Zoom Ratio to 1X...........................................................................1-28
1-2-7-3 Setting the Channels .......................................................................................1-28
1-2-7-4 Setting the Highest Scan Speed......................................................................1-29
1-2-7-5 Setting the XY Observation Mode ...................................................................1-30
1-2-7-6 Repeated Scanning Operation ........................................................................1-30
1-2-7-7 Setting the Cross-section to be Observed.......................................................1-31
1-2-7-8 Setting the Area to be Observed .....................................................................1-32
1-2-7-9 Setting a Lower Scan Speed ...........................................................................1-32
1-2-7-10 Stopping Repeated Scanning........................................................................1-33
1-2-8 Acquiring Image ..........................................................................................1-33
1-2-9 Saving Image .............................................................................................. 1-34
1-2-10 Exiting from the Software, Turning Power Off ........................................... 1-35
CONTENTS
1-3 Online Help..................................................................................... 1-36
2 APPLIED OPERATIONS 2-1
2-1 General Operation Procedure ........................................................2-1
1-3-1 Function Help ..............................................................................................1-36
1-3-2 Microscope Help..........................................................................................1-37
1-3-2-1 Configuring the Microscope ............................................................................ 1-40
1-3-2-2 Parfocality Correction and Jog Sensitivity Adjustment ................................... 1-46
1-3-2-3 Configuring the Filters (When using a filter system) ....................................... 1-52
1-3-2-4 Configuring the filters (When using a spectral detecting system)................... 1-56
1-3-2-5 Setting the C.A. Diameters ............................................................................. 1-59
2-1-1 Image Acquisition Procedure (Section (A))....................................................2-3
2-1-2 Image Acquisition Procedure in an Observation Mode (Section (B)) ............2-4
2-1-3 Examples of Operation Procedures...............................................................2-5
2-2 Image Acquisition............................................................................ 2-7
2-2-1 Image Acquisition in XY Observation Mode ..................................................2-8
2-2-1-1 Configuring the Microscope .............................................................................. 2-9
2-2-1-2 Setting the Filters ............................................................................................ 2-15
2-2-1-3 Setting the ND Filters...................................................................................... 2-17
2-2-1-4 Setting the Observation Condition .................................................................. 2-19
2-2-1-5 Acquiring Image .............................................................................................. 2-28
2-2-1-6 Acquiring Image in Accumulation Mode.......................................................... 2-29
2-2-1-7 Saving the Acquired Image in File .................................................................. 2-33
2-2-2 Image Acquisition in Other Observation Modes ..........................................2-34
2-2-2-1 XZ Observation Mode ..................................................................................... 2-34
2-2-2-2 XT Observation Mode ..................................................................................... 2-38
2-2-2-3 XZT Observation Mode................................................................................... 2-40
2-2-2-4 XYZ Observation Mode................................................................................... 2-46
2-2-2-5 XYT Observation Mode................................................................................... 2-51
2-2-2-6 XYZT Observation Mode ................................................................................ 2-55
2-2-3 Differences in Image Acquisition Method Between Fluorescent and Transmitted
CONTENTS
Images......................................................................................................... 2-61
2-2-3-1 Monochrome Image.........................................................................................2-61
2-2-3-2 Dual-Fluorochrome Image ...............................................................................2-64
2-2-3-3 Transmitted Image...........................................................................................2-67
2-2-4 Image Acquisition by Rotating It (Rotation Scan)........................................ 2-71
2-2-5 Image Acquisition of Only the Rectangular Position (Clip Scan)................. 2-72
2-2-6 Image Acquisition by Magnifying the Rectangular Position (Zoom-In Scan)2-75
2-2-7 High-Speed Image Acquisition .................................................................... 2-78
2-2-8 Image acquisition to prevent crosstalk between fluorescence (Sequential Scan)
.................................................................................................................... 2-79
2-2-8-1 Virtual Channel Function .................................................................................2-82
2-2-9 Image Acquisition of a Line at Desired Angle.............................................. 2-88
2-2-10 Display the change of image intensity (Point Scan) .................................. 2-89
2-2-11 Image Acquisition on Desired Line (XZ, XT or XZT Observation)............. 2-94
2-2-12 Image Acquisition in the Laser Excitation Mode........................................ 2-97
2-2-12-1 Making REX Mask File ..................................................................................2-98
2-2-12-2 Example of FRAP experiment .....................................................................2-103
2-2-13 Notes for image acquisition ..................................................................... 2-112
2-2-13-1 Memory of setting information for scanning region......................................2-112
2-3 Saving, Opening and Shredding Images................................... 2-113
2-3-1 Saving Images........................................................................................... 2-116
2-3-1-1 Saving Images As a Series............................................................................2-116
2-3-1-2 Saving a Display ............................................................................................2-118
2-3-1-3 Saving Specified Area of Image ....................................................................2-120
2-3-1-4 Saving Animation Images ..............................................................................2-123
2-3-1-5 File Types Available for Save ........................................................................2-125
2-3-2 Opening Previously Saved Images ........................................................... 2-132
2-3-3 Shredding Images .....................................................................................2-133
2-3-4 Saving Comment Together with Image ..................................................... 2-135
2-3-5 Checking the Image Information/Acquisition Parameters.......................... 2-138
2-3-6 Saving the Image Information/Observation Condition............................... 2-142
2-3-7 Saving/Reading the Region File................................................................ 2-145
CONTENTS
2-4 Protocol processor...................................................................... 2-150
2-3-7-1 Saving the Region File.................................................................................. 2-145
2-3-7-2 Reading the Region File ............................................................................... 2-147
2-4-1 Starting the Protocol Processor.................................................................2-151
2-4-2 Editing the Protocol ...................................................................................2-152
2-4-2-1 Description of Setting Items .......................................................................... 2-153
2-4-2-2 Command List ............................................................................................... 2-160
2-4-2-3 Protocol Repetition Processing..................................................................... 2-165
2-4-2-4 Input supporting function............................................................................... 2-167
2-4-2-5 Protocol procedure supporting function ........................................................ 2-169
2-4-3 Saving the Protocol....................................................................................2-170
2-4-4 Executing the Protocol...............................................................................2-172
2-4-5 Loading a Protocol.....................................................................................2-174
2-4-6 Loading a protocol of previous format .......................................................2-175
2-4-7 COM Communication function................................................................... 2-177
2-4-8 Example of assembling a protocol.............................................................2-178
2-4-8-1 Example of Setting Procedure of FRAP Observation ................................... 2-178
2-4-8-2 Example of Setting Procedure of XYT Observation for hours (Protocol using
repetition processing) ........................................................................................... 2-187
2-4-9 Pop-up Menu .............................................................................................2-195
2-4-10 Restrictions of [PAPP] setting.................................................................. 2-196
2-4-10-1 Restrictions with [Mode] and [Sub Mode] ................................................... 2-196
2-5 Changing the Image Display Method......................................... 2-198
2-5-1 Displaying an Image in Simulated Colors .................................................. 2-198
2-5-2 Editing the LUT (Look Up Table) ...............................................................2-200
2-5-2-1 LUT Graph Editing According to Colors........................................................ 2-200
2-5-2-2 LUT Graph Editing by Gamma Correction.................................................... 2-202
2-5-3 Switching the Displayed Channels (Ch1 – Ch5)........................................2-203
2-5-4 Displaying Images of Multiple Channels Simultaneously (Side By Side Views,
Over And Under Views, Single View) ........................................................2-204
2-5-4-1 Displaying Images Separately Per Channel (Side By Side Views, Over And Under
Views) ................................................................................................................... 2-205
CONTENTS
2-5-4-2 Displaying Merged Image of Multiple Channels (Single View)......................2-206
2-5-5 Changing the Number of Divided Images ................................................. 2-208
2-5-5-1 Increasing the Number of Divided Images ....................................................2-208
2-5-5-2 Decreasing the Number of Divided Images...................................................2-210
2-5-6 Switching the Display Method of Multiple Images ..................................... 2-211
2-5-7 Displaying Multiple Image Slices Together ............................................... 2-213
2-5-7-1 Displaying Multiple Images Per Channel.......................................................2-214
2-5-7-2 Displaying Images of Two Channels Together..............................................2-216
2-5-7-3 Displaying Time-Lapse Images .....................................................................2-217
2-5-7-4 Displaying Multiple Multiple sections Images ................................................2-218
2-5-7-5 Displaying Same Images in Different Display Methods.................................2-218
2-5-7-6 Re-arranging Images Using the Same Display Method ................................2-221
2-5-7-7 Displaying Different Images Together ...........................................................2-221
2-5-8 Magnifying/Reducing an Image................................................................. 2-224
2-6 Image Processing ........................................................................ 2-225
2-6-1 Filtering...................................................................................................... 2-225
2-6-1-1 Contour Enhancement...................................................................................2-226
2-6-1-2 Noise Reduction ............................................................................................2-229
2-6-1-3 Image Sharpening .........................................................................................2-230
2-6-1-4 DIC Correcting DIC Level Irregularities .........................................................2-231
2-6-2 Contrast Conversion.................................................................................. 2-233
2-6-3 Mathematical Operations Between Images............................................... 2-236
2-6-3-1 Image Addition...............................................................................................2-236
2-6-3-2 Image Subtraction..........................................................................................2-239
2-6-3-3 Image Multiplication .......................................................................................2-239
2-6-3-4 Image Division ...............................................................................................2-240
2-6-3-5 NOT Image ....................................................................................................2-241
2-6-3-6 Image AND Image .........................................................................................2-243
2-6-3-7 Image OR Image ...........................................................................................2-244
2-6-3-8 Image XOR Image .........................................................................................2-244
2-6-4 Brightness overlap level between 2 channels (Colocalization).................. 2-246
2-6-4-1 Annotation Mode............................................................................................2-247
CONTENTS
2-7 Image Analysis............................................................................. 2-274
2-6-4-2 Colocalization for series image data set ....................................................... 2-254
2-6-4-3 Image measurement ..................................................................................... 2-255
2-6-5 Appending image (Append).......................................................................2-258
2-6-5-1 Appending two images.................................................................................. 2-258
2-6-5-2 Appending image from several image data set ............................................ 2-262
2-6-6 Extract image (Crop)..................................................................................2-267
2-7-1 Checking the Intensity of a Specific Part ...................................................2-275
2-7-1-1 Intensity Values on a Line (Line Profile) ....................................................... 2-275
2-7-1-2 Intensity Values on a Planar Region (Bird’s Eye View) ................................ 2-278
2-7-2 Checking the Intensity Distribution of a Specific Part ................................2-283
2-7-2-1 Intensity Distribution on a Line (Histogram) .................................................. 2-283
2-7-2-2 Intensity Distribution on a Planar Region (Histogram).................................. 2-284
2-7-3 Image Measurement..................................................................................2-286
2-7-3-1 Length Measurement .................................................................................... 2-286
2-7-3-2 Area Measurement........................................................................................ 2-286
2-7-3-3 Measuring the Change in Mean Value of Intensity ....................................... 2-287
2-7-3-4 Measuring the Change in Integrated Intensity .............................................. 2-293
2-8 Building an Image from a Different Viewpoint.......................... 2-295
2-8-1 Building Extended Focus Image from XYZ Image.....................................2-295
2-8-1-1 Display Switching to Built Image................................................................... 2-295
2-8-1-2 Turning Built Image into Single Image .......................................................... 2-298
2-8-1-3 Turning Built Image into time series image................................................... 2-300
2-8-2 Building line images to be viewed in Z direction........................................2-301
2-9 Viewing 3D Image ........................................................................2-305
2-9-1 Successive Display of Images...................................................................2-307
2-9-1-1 Changing the Successive Display Speed ..................................................... 2-308
2-9-1-2 Changing the successive image display position ......................................... 2-309
2-9-2 Animation...................................................................................................2-310
2-9-3 Building Stereo 3D Images........................................................................2-312
2-9-4 Building a 3D Image to be Viewed Through Color (Red/Green) Eyeglasses2-314
2-10 Viewing Images Following the Progress of Time................... 2-316
CONTENTS
2-10-1 Displaying Images Together.................................................................... 2-316
2-10-2 Displaying Images Successively .............................................................2-316
2-11 Transferring Data to Another Application...............................2-319
2-11-1 Transferring Analysis Data to Another Application.................................. 2-319
2-11-2 Transferring the Plot Image of Analysis Data to Another Application...... 2-322
2-11-3 Transferring Image Data to Another Application (Paint, etc.).................. 2-323
2-12 Entering Comment in Image .....................................................2-324
2-12-1 Writing Characters in Image.................................................................... 2-324
2-12-2 Displaying the Image Intensity ................................................................ 2-326
2-12-3 Displaying the X-coordinate/Y-coordinate of the Image .......................... 2-327
2-12-4 Drawing a Figure in Image ...................................................................... 2-328
2-12-5 Drawing a Scale in Image ....................................................................... 2-329
2-12-6 Drawing an Arrow in Image ..................................................................... 2-330
2-12-7 Drawing Color Bars in Image .................................................................. 2-331
2-12-8 Deleting Comment................................................................................... 2-332
2-12-9 Moving Comment ....................................................................................2-333
2-12-10 Changing the Comment Size ................................................................ 2-334
2-12-11 Changing the Comment Color............................................................... 2-335
2-12-12 Changing the Comment Font ................................................................ 2-336
2-13 Image Output at Printer.............................................................2-337
2-14 Merger/Extraction of Image Channels ..................................... 2-338
2-14-1 Setting the Range of Multiple Image Slices............................................. 2-338
2-14-2 Merging Image Channels ........................................................................ 2-339
2-14-3 Extracting Channels from Image ............................................................. 2-344
2-15 Changing the Chart Display Method........................................ 2-348
2-15-1 [Chart] Panel ...........................................................................................2-348
2-15-2 [Series] Panel .......................................................................................... 2-356
2-16 Pop-up Menus ............................................................................ 2-359
Appendix A List of Hot Keys A-1
CONTENTS
Appendix B Glossary B-1
Appendix C USER REGISTRATION OF FV1000 C-1
Appendix C-1 User Registration.......................................................... C-1
Appendix C-2 Logging into the FV1000.............................................. C-4
Appendix C-3 Deleting a User ............................................................. C-5
Appendix D Change of Default Folder for [File I/O] Panel
D-1
Appendix E List of Functions in the [Active Overlays]
Dialog Box E-1
Appendix E-1 Coordinate Position Data..............................................E-1
Appendix E-1-1 X-Coordinate ................................................................................ E-1
Appendix E-1-2 Y-Coordinate ................................................................................ E-2
Appendix E-1-3 Other ............................................................................................ E-3
1 Z Position....................................................................................................................E-3
2 T Position....................................................................................................................E-3
3 Animation.................................................................................................................... E-3
Appendix E-2 Intensity Data .................................................................E-4
Appendix E-3 Other ...............................................................................E-5
Appendix E-3-1 Channel Number .......................................................................... E-5
Appendix E-3-2 Objective Power ........................................................................... E-5
Appendix E-3-3 Date of Image Capturing .............................................................. E-5
Appendix E-3-4 Time of Image Capturing.............................................................. E-5
Appendix E-3-5 Image File Name .......................................................................... E-5
Appendix F Hand Switch and Microscope Frame Function
CONTENTS
Allocation F-1
Appendix F-1 Hand Switch Functions .................................................F-1
Appendix F-1-1 BX/BXWI .......................................................................................F-1
Appendix F-1-2 IX...................................................................................................F-2
Appendix F-2 Microscope Frame Functions .......................................F-3
Appendix F-2-1 BX .................................................................................................F-3
Appendix F-2-2 IX...................................................................................................F-4
Appendix F-2-3 Focus Adjustment Knob ................................................................F-5
1 BX............................................................................................................................... F-5
2 IX ................................................................................................................................ F-5
Getting Started FLUOVIEW/ Basic Operations
r
r
)

1 Getting Started FLUOVIEW

1-1 Basic Operations
1-1-1 Microscope
The following figure shows the major controls of a microscope. The actual configuration of
the modules including the specimen stage, revolving nosepiece and lighting equipment may
differ from those shown below.
For detailed microscope operation procedures, refer to the instruction manuals of your
microscope.
Combination with BX61
(1) Light path
selector knob
(2) Cube turret /Cube display window
(This figure shows Cube display
window)
(4) Transmitted light DIC
FV5-DICTS/ WI-DICTH (with BX61WI) (optional)
(3) Analyze
IX2-MDICT
(6) Universal
condense
(This figure shows the case of BX)
(5) Filters
LBD ND6
ND25
When using BX61WI,
LBD
FFR
(7)Hand switch
U-HSTR2
(U-FH is optionally available with BX61
OPERATION INSTRUCTIONS
Page
1-1
Getting Started FLUOVIEW/Basic Operations
2
(1) Light path selector knob
Select the light path between the visual observation and photography observation.
See the following table and set the knob to the position corresponding to the
required light path.
Light path selector knob Symbol Intensity Ratio
Pushed in 100% for the visual observation
Middle position
Pulled out 100% for photography observation
(2) Cube turret
Select the fluorescence observation tube by rotating the turret.
20% for the visual observation, 80% for photography observation
Engage the desired cube in the light path for visual fluorescence observation or
visual transmitted light observation.
For laser microscopy, rotate the turret to page tab
state that mirror cube is entered.)
(3) Analyzer IX2-MDICT
Polarizing plate for use in differential interference observation and polarized light
observation.
Rotate the turret to engage the IX2-MDICT in the light path for visual transmitted
light differential interference observation or transmitted polarized light observation.
(4) Transmitted light DIC slider U-DICTS/WI-DICTHRA (built-in FV10-SRE or FV10-
SNPXLU of BX61WI)
This is the prism for use in differential interference observation.
Engage the transmitted light DIC in the light path for laser differential interference
observation or visual transmitted light differential interference observation.
Leaving the transmitted light DIC engaged during laser fluorescence observation will
1
. (Set cube turret in the
degrade the resolution somewhat. We recommend disengaging the transmitted light
DIC from the light path when simple laser fluorescence observation is required.
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Getting Started FLUOVIEW/Basic Operations
NOTE
(5) Filters
These filters are used to adjust the transmitted light.
Be sure to disengage any filter from the light path for transmitted observation using
lasers. Leaving a filter engaged in the light path will degrade the image quality.
When you perform transmitted observation using laser with BX61WI, use the filter
knob to disengage the LBD from the light path and engage the FR (Frost) into the
light path. Disengaging the FR (Frost) from the light path may generate interference
fringes on an image.
(6) Universal condenser
With transmitted light differential interference observation using an
immersion objective, set the microscope’s field diaphragm so that
it circumscribes the field of view. Otherwise the contrast may
degrade. (This applies to both visual observation and laser
differential interference observation.)
Condenser for transmitted lighting. In addition, the rotary turret for the transmitted light
DIC prism and the polarizing plate for differential interference observation (polarizer)
are also provided.
To perform differential interference observation, engage the transmitted light DIC
prism matching the objective in use in the light path (For both visual observation and
laser differential interference observation).
To perform visual differential interference observation or laser differential
interference observation, engage the polarizing plate in the light path.
(7) Hand switch U-HSTR2 (U-FH is optionally available with BX61/BX61WI)
This is the hand switch to operate the BX motorized system.
NOTE
Connect the filter wheel of BX to connector of the following in the
back of UCB.
FW0: FW1
FWR: FW2
FWT: FW3
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y
)
r
Combination with IX81 FVF
(4) Condense
(5) Filters
(6) Light path selector button
(1)Fluorescence mirror unit
((2) Analyzer IX-MDICT
Optionall
installed
(4) Transmitted light
DIC slider
U-DICTS (optional)
(6)Hand switch
U-HSTR2
(U-FH is optionally available.)
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(1) Fluorescence mirror unit
Select the fluorescence observation tube by rotating the turret.
Engage the desired cube in the light path for visual fluorescence observation.
For laser microscopy, rotate the turret to page tab
mirror cube is entered.)
(2) Analyzer IX2-MDICT
Polarizing plate for use in differential interference observation and polarized light
observation.
Rotate the cube turret to engage the IX2-MDICT analyzer into the light path for visual
transmitted light differential interference observation or transmitted polarized light
1
. (Set turret in the state that
observation.
(3) Transmitted light DIC slider U-DICTS
This is a prism for use in differential interference observation.
Engage U-DICTS in the light path for laser differential interference observation or
visual transmitted light differential interference observation.
Leaving U-DICTS engaged during laser fluorescence observation will degrade image
quality somewhat. We recommend disengaging the U-DICTS from the light path
when simple laser fluorescence observation is required.
NOTE
With transmitted light differential interference observation using an
immersion objective, set the microscope’s field diaphragm so that it
inscribes the field of view. Otherwise the contrast may degrade.
(This applies to both visual observation and laser differential
interference observation.)
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6
(4) Condenser, polarizing plate
Condenser for transmitted lighting.
In addition, the rotary turret for the transmitted light DIC and the polarizing plate for
differential interference observation (analyzer) are also provided.
To perform differential interference observation, engage the transmitted light DIC
(optional) matching the objective in use in the light path (For both visual observation
and laser differential interference observation).
To perform visual differential interference observation or laser differential
interference observation, engage the polarizing plate in the light path.
(5) Filters
These filters are used to adjust transmitted light.
For transmitted observation using laser, disengage the LBD filter from the light path
and engage the FR (frost) filter in the light path by operating the filter levers. If the FR
filter is disengaged from the light path, the image may suffer from stripe interference.
(6) Light path selector button
Select the light path between the visual observation and photography observation.
When <
When <
(7) Hand switch (U-FH is optionally available.)
This is the hand switch to operate the IX motorized system.
> LED is lighted, visual observation can be done.
> LED is lighted, TV or photography observation can be done.
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1-1-2 General Mouse Operation Procedures
Use the mouse to select a command, character string or button. Use the left
button of the mouse unless otherwise specified.
To select or execute something: Clicking
To click the mouse, place the mouse pointer on the desired function and
press the mouse button once.
(Pressing the right button of the mouse is referred to as right-clicking.)
To select something and execute its function: Double clicking
To double-click, place the mouse pointer on the desired function and press the
Getting Started FLUOVIEW/Basic Operations
mouse button successively twice.
To move something: Dragging
To drag, place the mouse pointer on the desired function, and while pressing
and holding the mouse button, move the mouse to the desired destination. At
the desired destination, release the mouse button.
(Dragging by pressing the right button of the mouse is referred to as right-
dragging.)
One Point!
When the mouse is moved, the picture of arrow on the screen moves
accordingly. The picture which moves on the screen as the mouse is
moved is referred to as the mouse pointer.
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f
r
g
r
f
1-1-3 Names of Major Panel and Window Controls and Their Functions
The window as shown below is displayed when FLUOVIEW starts up. FLUOVIEW uses
panel-type windows.
This section describes the names of the major controls displayed in panels and windows.
Click to turn the window into an icon.
Original size button <
Click each button to
execute the processing
Drop-down list
Click the list of available
items for selection.
To select an item in
the list, click the item.
Buttons
indicated on it.
to display
Control menu box
Clicking this box displays a control menu, which contains the commands for use in controllin
the window.
Click to return the maxim-size window to its original size.
Title bar
Shows the title of the window. The title ba of a window that is active is displayed in a
different color from that of other windows.
Information
Shows information on the operations and meanings of functions.
Scale
The scale is used to set a value which is continuously variable in a certain range. Clicking a point in the scale area allows the value to change 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.
Option buttons
This is a group of multiple items among which only one can be selected. Clicking one of the round buttons selects the corresponding item. The option button of the selected item is displayed with a black dot in the center o
Minimize button < >
>
Sub-panel
A sub-panel is
provided for use in
detailed setting o
information display o
a function.
Fig. 1-1 Window and Major Functions
Check box
Clicking this box enables or disables the indicated item. The item is enabled when the check box is checked (X).
Scroll bar
The scroll bar is displayed when there are too many data items to be displayed in a field at once, and is used to display the data items outside the field. Clicking a point in the scroll area allows data items to be scrolled in large steps. Clicking the top or bottom arrow button allows fine scrolling of the data items. Dragging the square knob allows direct scrolling.
List box
Shows the list of available items for selection. All items in the list can be displayed by scrolling. To select an item in the list, double-click or drag the item.
Fig. 1-2 Sub-panel and Major Functions
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Group box
The group box groups functions with specific meanings and encloses frame.
them in a
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