Olympus FLUOVIEW FV1000 User Manual

User’s Manual

FLUOVIEW

FV1000

CONFOCAL LASER SCANNING

BIOLOGICAL MICROSCOPE

Petition

This user’s manual is for the 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.

OVERVIEW”, “PREPARATION FOR OPERATION” and “TROUBLE Q&A”. Together
with this manual, please also read the “SAFETY 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.

This user’s manual is composed of three volumes including “SYSTEM

[HARDWARE]

AX7282

CAUTION

CAUTION

1. Reproduction, copying or duplication of a part or all of this software and manual is prohibited.

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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MANUAL CONFIGURATION

MANUAL CONFIGURATION

I. SYSTEM OVERVIEW

1. System Overview............................................................................................I. 1-1

II. PREPARATION For OPERATION

1. Preparation for Operation...............................................................................II. 1-1

2. Replacement of Cubes...................................................................................II. 2-1

3. Centration of Mercury Burner .........................................................................II. 3-1

III. TROUBLE Q&A

1. Troubleshooting Guide ...................................................................................III. 1 -1

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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.

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This volume describes the overview of the FLUOVIEW
FV1000 system.
Please read this volume so that you can understand the
system before use.

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CONTENTS

1 System Overview 1-1

1-1 Principles䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-1
1-2 Features of the FV1000 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-2

1-3 Optical Path Diagram

䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-3

1-4 System Configuration 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-4

1-4-1 System Diagram 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-4

1-4-2 System Appearance and Functions 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-5

System Overview / Principles

1 System Overview

OLYMPUS FV1000 is a confocal laser scanning biological microscope system featuring improved basic

performances (sensor system, scanning system and illumination system performances) by considering the “live

cell observations”, with which long hours of stable measurement of weak fluorescence is required.

This microscope is equipped with 3 fluorescence channels, 3 lasers and AOTF to meet various applications in a

wide range of advanced research fields.

1-1 Principles

A laser scanning microscope converges the laser beam into a small spot using an

objective and scans the specimen in the X-Y direction using the laser beam.

The microscope then captures the fluorescent light and reflected light from the specimen

using light detectors and outputs the specimen image on an image monitor.

As shown in this figure, the confocal

optics incorporates a confocal aperture

on the optically conjugate position

(confocal plane) with the focus position to

eliminate light from other part than the

focus position. This causes the

extraneous light to be viewed as

darkness in the observation image, it is

possible to slice optically a tissue

specimen that has thickness.

On the other hand, an ordinary optical

microscope, the light from other part than

the focus position is overlapped with the

imaging light of the focus position so the

image is blurred in overall.

The laser beam that has transmitted

through the specimen is detected by the

transmitted light detector and provides

the transmitted image, which is not a

confocal image.

However, when the fluorescence images

of the transmitted and confocal images are combined, it is possible to obtain very

important information on the specimen.

Light detector

Confocal
aperture

Objective

Specimen

Light detector

Laser

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I. System Overview

I. 1-1

System Overview / Features of the FV1000

1-2 Features of the FV1000

1. The photon counting mode is newly provided to improve the sensitivity and S/N and

to enable quantitative optical intensity measurement. Photon counting makes

possible long hours of quantitative observation by completely eliminating analog-

derived drift. The dynamic range in which photon counting is possible is expanded

using a newly designed wideband head amplifier and processing circuitry.

2. High-speed imaging at 8 frames per sec. is made possible by fast galvano mirror. In

addition, high-speed image acquisition is possible without stopping the Z-series

motors used in the XYZ and XZ observations.

3. During long hours of time-lapse observation, a stable supply of excitation light is

made possible thanks to the feedback control of the intensity of each laser.

Together with the photon counting function, this function ensures the stability and

quantitative nature of long-hour observations.

4. Three fluorescence channels, three lasers and AOTF are provided as standard to

meet a large variety of applications.

5. With a fully-motorized scan unit and motorized microscope, the entire system is

motorized so the scanning conditions including those of the optics can be saved

and reproduced.

6. When an extension laser irradiation unit is used for photon activation aiming at

causing discoloration, optical simulation or uncaging of the specimen, a system

optimized for cell function analysis experiments can be built.

7. When the system incorporates the spectral detector unit that is composed of a 2-

channel spectral detector and 1-channel filter, it is possible to set the detection

conditions more flexibly, acquire the fluorescence spectral data and use the

fluorescence isolation function.

I. System Overview

I. 1-2

Page

1-3 Optical Path Diagram

System Overview / Optical Path Diagram

Laser beam

Galvano mirror

VIS

Ch3

UV

Ch4

Barrier filter

IR

Barrier filter

Grating

Ch2

DM

Slit

Ch1

DM

DM

Pinhole

Grating

Slit

Mercury light Supply

Galvano mirror

DM

Page

I. System Overview

I. 1-3

System Overview / System Configuration

I. System Overview

I. 1-4

Page

1-4 System Configuration

1-4-1 System Diagram

Swinging Nosepiece

Microscope

XLU Single-Position
Nosepiece

Anti-

vibration Table

Non-confocal Point

Detector for BX

Epi Fiber Illumination

Unit

Scan Unit
for BX

Scan Unit
for BXWI

Mercury Lamp Housing

Spectral Fluorescent Detector

Filter Type Fluorescent

Transmitted Light Detector

Non-Confocal Detector for SU

Additional Scan Unit

Power Supply Unit

Microscope

Air Anti-Vibration

Tab]e

Scan Unit
for IX

Mercury lamp Housing

Additional 4

th

Channel

Fluorescent Sensor

Fiber Port for Fluorescent Detection

Non-Confocal
Detector for IX

Epi Fiber

Illumination Unit

Control Box for

Microscope

Power Supply Unit

VIS laser Fiber Unit

LD405/440 Laser

Laser Combiner

Argon laser

HeNe Green Laser

HeNe Red Laser

System Controller

PC Interface Board

20’ Flat Panel Display

Basic Software

Time Course Software

Review Station Software

Advanced software

Advanced

Filter Set

for SPD MBGR

Filter Set

for FD

MBGR

1-4-2 System Appearance and Functions

r

A

r

A

y

The applicable microscopes are the BX61/62TRF, BX61WIF and IX81F.

Epi Fibe

Illumination Unit

㪝㪠㪩

Microscope

Top View

System Overview / System Configuration

Laser Power Supply

Mouse

Keyboard

Epi Fiber Illumination Unit

Illumination unit based on

mercury burner, connected to

the microscope through a fiber.

Scan Unit

Heart of lase

scanning

microscope, composed

of scanner and light detector.

Microscope

Designed for fluorescence

observations.

Anti-vibration table

Control box

Controls

the microscope.

Mercury power suppl

Power supply for reflected

light illumination.

Microscope

UCB

RFL

VT

Power supply unit

lso controls the FV1000 scan

unit, laser combiner, etc.

20’ Flat Panel Display

Monitors for displaying
the laser scanning image
and control panel, etc.

LCD LCD

PSU

㩷

TD

Transmitted light detector

(incl. transmitted light illumination unit)

Unit for obtaining the transmitted
image, connected to the
microscope through a fiber.

COMB

Power supply

Power supply

Keyboard

Laser combiner

Unit for combining the laser light
of the Ar, HeNe green and
HeNe lasers into a single fiber.

Laser power supply

Power supply for the
Ar, HeNe green
and HeNe red lasers.

System Controller

Used to control
the FV1000, file
its images, etc.

PC

Front View

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I. System Overview

I. 1-5

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This volume describes the methods for preparation for
operation of the FLUOVIEW FV1000 system.
After completing the preparations, activate the software and
start observation by controlling the display on the monitor
screen.
Please read this volume so that you can understand the
system before use.

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CONTENTS

1 Preparation for Operation 1-1

1-1 Turning the Power On 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-1
1-2 Starting the Software 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-3
1-3 Exiting from the Software 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃

䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-4

1-4 Turning the Power Off 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 1-4

2 Replacement of Cubes 2-1

2-1 Replacing the DM Cube 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-1

2-1-1 With the FV10-ASU 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-1

2-1-2 With the FV10-OPD 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-3

2-2 Replacing the Spectral Cube 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-4

2-2-1 Removing the spectral cube 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-4

2-2-2 Fabricating a spectral cube 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-6

2-2-3 Attaching the spectral cube 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 2-6

3 Centration of Mercury Burner 3-1

3-1 Centering the Mercury Burner䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 3-1

Preparation for Operation / Turning the Power On

1 Preparation for Operation

1-1 Turning the Power On

1. Set the power switches of the following units to I (ON).

x Power Supply Unit FV10-PSU

x Mercury Burner Power Supply Unit

x Microscope Control Box BX-UCB or IX2-UCB

2. Set the power switches of the PC and monitor to I (ON).

3. Turn on the lasers as follows.

䇭䇭Ar laser power supply (Type 300)

HeNe Green/Red laser power supply

c

㽲

㽳

3.1䇭Argon laser: FV5-LA-MAR

x Set the power switch to ON. (This starts the fan of the laser.)

x Turn the key to the ON position.

It takes a few tens of seconds after the key is set to ON till the

laser oscillation begins.

3.2䇭Helium-Neon Green/Red Laser: FV5-LA-HEG/HER

x Turn the key to the I (ON) position.

It takes a few tens of seconds after the key is set to ON till the

laser oscillation begins.

NOTE

To ensure stable laser light output, it is recommended to

warm up the laser power supply after turning it on. The

warm-up period should be 10 minutes or more when using

the Argon laser power supply and 30 minutes or more when

using the Helium-Neon Green or Red laser power supply.

II. PREPARATION For OPERATION

Page

II. 1 - 1

Preparation for Operation / Turning the Power On

3.3䇭LD405/440 laser: FV10-LD405/440

f

d e

c

LD405/440 laser power supply

NOTE

x Make sure that the provided shorting plug is attached

to the remote interlock c or that is connected to your

equipment and the interlock is released.

x Set the power switch d to ON.

x Turn the key e to the ON position.

x Set the shutter switch f OPEN.

The red lighting of the LASER EMISSION LED of

the LD405/440 laser power supply indicates that

the laser is oscillating. With a certain setup, the

laser beam is output by simply setting the shutter

switch f to ON.

II. PREPARATION For OPERATION

II. 1 - 2

Page

1-2 Starting the Software

Preparation for Operation / Starting the Software

FV10-ASW

[FLUOVIEW] icon

FV10-SW

NOTE

NOTE

1. Enter the user name and password to log in the Windows.

NOTE

2. Double-click the [FLUOVIEW] icon on the desktop.

TIP

Turn on the microscope and power supply units before starting this

software.

If you are [FV10-SW] user, don’t double click the [FLUOVIEW Setup]

icon on the desktop and boot it up while the FLUOVIEW software is

running.

Log in using the user name given the Administrator’s authority.

If more than one user uses the FV1000, each user should personally log in

personally. For details, refer to Appendix E, “USER REGISTRATION OF

FV1000” in Volume [OPERATION INSTRUCTIONS].

[FLUOVIEW] icon

TIP

NOTE

It takes 20 to 30 seconds after the [FLUOVIEW] icon is double-clicked till

the software starts up.

Images cannot be observed if the manual shutter of the fluorescence

mirror unit is close. In this case, slide the shutter to the open

position.

II. PREPARATION For OPERATION

Page

II. 1 - 3

Preparation for Operation / Exiting from the Software

1-3 Exiting from the Software

Exit from the application software and shut down Windows.

NOTE

1-4 Turning the Power Off

Set the power switches of the units to O (OFF).

When using of Argon laser, Multi-line Argon laser

Turn the key to OFF position and wait for the fan to stop automatically when the laser

unit has cooled down. It takes several minutes until the fan of laser stops. Set the power

switch to OFF. (Also the power supply fan will stop automatically.)

䋨For details, refer to the instruction manual of laser units䋩

After exiting the application software, the light of mercury burner

power supply unit may exposure to specimen. To avoid this, perform

either of the followings,

· Close the manual shutter of the mercury burner power supply unit.

· Turn off the mercury burner power supply unit.

· Close the manual shutter of the fluorescence mirror unit(BX61WI or

IX81䋩.

II. PREPARATION For OPERATION

II. 1 - 4

Page

Replacement of Cubes / Replacing the DM Cube

2 Replacement of Cubes

2-1 Replacing the DM Cube

The DM cube is used to connect the light path of the optional FV10-ASU Auxiliary Scan

Unit or FV10-OPD Non-confocal Point Detector with that of the scan unit, and should be

selected according to the observation method.

2-1-1 With the FV10-ASU

1. Set the light path of the scan unit to the LSM light path. (This can be done with the

FLUOVIEW software. For details, refer to the User’s Manual for the FLUOVIEW

software.)

2. Loosen the four cover clamping knobs c on the lower part of the right side panel of

the scan unit, and remove the cover d.

d

c

3. Using an Allen screwdriver, loosen the screw f retaining the guide lock plate e,

move the guide lock plate in the direction of the arrow, engage it with the pin g

below the guide, and tighten the screw f again to lock the guide.

e

g

f

Before moving After moving

II. PREPARATION For OPERATION

Page

II. 2 - 1

Replacement of Cubes / Replacing the DM Cube

4. Using the Allen screwdriver, loosen the clamping screw h retaining the DM cube.

h

5. Pull out the DM cube insertion knob i toward you and take out the DM cube from

the light path selector mechanism.

i

6. Insert the desired DM cube in the dovetail of the light path selector mechanism, and

tighten the clamping screw h using the Allen screwdriver.

7. Loosen the screw f retaining the guide lock plate, slide it in the direction of the

arrow and tighten the screw f again.

8. Attach the scan unit cover to the original position.

II. PREPARATION For OPERATION

II. 2 - 2

Page

2-1-2 With the FV10-OPD

1. Perform the same operations as steps 1 to 5 in section 2-1-1, “With the FV10-ASU”

2. Using a precision Phillips screwdriver, loosen the screw clamping the DM holder

Replacement of Cubes䋯Replacing the DM Cube

to take out the DM cube.

plate and take out the DM and DM holder plate.

NOTE

NOTE

1.0



3.0

The applicable DM (Dichroic Mirror) diameter is

thickness of

The DM should be inserted by distinguishing the face and back. Make sure that

the reflective surface (interference film surface) of the DM comes as the face.

.

mm05.01r



u

1.0



3.0



3826

mm

, with

3. Insert the desired DM and tighten the screw to clamp the DM holder plate.

II. PREPARATION For OPERATION

Page

II. 2 - 3

Replacement of Cubes / Replacing the Spectral Cube

2-2 Replacing the Spectral Cube

To improve the efficiency of fluorescence detection, the fluorescence waveform

separating dichroic mirrors and barrier filters (2 channels) of the spectral cube inside the

external photo-multiplier (FV10-OPD) can be replaced according to the excitation

wavelength to be used.

2-2-1 Removing the spectral cube

1. Using an Allen screwdriver, loosen the two cover clamping screws (provided with

slip-off prevention mechanisms) c on the left side panel of the FV10-OPD and

remove the cover d.

2. Loosen the cube cover clamping screws e inside the cover in the same way as in

step 1, and remove the cube cover by holding the cover knob f.

d

c

e

f

II. PREPARATION For OPERATION

II. 2 - 4

Page

Replacement of Cubes䋯Replacing the Spectral Cube

3. Loosen the spectral cube clamping screw g a little using the Allen screwdriver, and

pull out the spectral cube by holding the spectral cube insertion knob h.

g

h

II. PREPARATION For OPERATION

Page

II. 2 - 5

Replacement of Cubes / Replacing the Spectral Cube

r

*

*

r

2-2-2 Fabricating a spectral cube

A desired spectral cube can be fabricated by attaching a commercially available barrier

filter and DM to the spectral cube frame.

Dimensional conditions for the optical components

1.0



2.0

Barrier filter mm

Dichroic mirror mm

Filter holder

ring

Barrier filte

DM holde

I

DM



25

1.0



3.0



,䇭max. thickness mm6

1.0



3.0



3826

u , thickness mm05.01r

Clamping screw (cross-head)

Barrier filter

Filter holder ring

Reflective surface
(interference film
surface)

NOTE

When replacing the DM and barrier filter, take special care not to contaminate

them with fingerprints, etc.

* Orientation of the filter holder ring

NOTE

Attach the filter holder ring by changing its orientation according to the filter

thickness, insert the tip of a precision flat-blade screwdriver into the notch on the

ring and turn the ring taking care not to scratch the filter to lock it.

䇭䇭䇭䇭䇭䇭䇭䇭䇭

2-2-3 Attaching the spectral cube

Attach the spectral cube, together with the DMs and barrier filters, by reversing the

removing procedure.

II. PREPARATION For OPERATION

II. 2 - 6

Page

Filter thickness 4-6 mm Filter thickness <4 mm

Centration of Mercury Burner / Centering the Mercury Burner

3 Centration of Mercury Burner

3-1 Centering the Mercury Burner

For the reflected light fluorescence observation, refer to the User’s Manual for the

Reflected Light Fluorescence System.

Since this system introduces the light of a mercury burner through the light guide, the

burner centering method is slightly different from that described in the User’s Manual for

the Reflected Light Fluorescence System. This section is intended to describe the

method specific to this system.

TIP

NOTE

1. Turn the shutter c fully toward the bottom to block the light.

When the light guide is disconnected, another built-in shutter is engaged

automatically in the light path to ensure safety.

2. Remove the light guide d from the ULH holder and replace with the centering

target e.

3. Turn the shutter c toward the open direction. The arc image of the mercury burner

will be visible on the screen f of the centering target e.

The arc image stabilizes in 5 to 10 minutes after the mercury burner is turned on.

Keep the shutter c closed except for centering operation to prevent the centering

target from being heated up.

II. PREPARATION For OPERATION

Page

II. 3 - 1

Centration of Mercury Burner / Centering the Mercury Burner

4. Turn the collector lens focusing knob g on the lamp housing to bring the arc image

into focus.

TIP

TIP

Hereafter, centering is possible with the method described in the User’s Manual for the

Reflected Fluorescence System, that is, using the burner centering knob and mirror

focusing screw.

5. After completing centering, remove the centering target and connect the light guide.

When starting observation, turn the collector lens focusing knob g to maximize the

brightness of the observation field.

The mercury burner does not have to be centered until the next time it is replaced.

II. PREPARATION For OPERATION

II. 3 - 2

Page

T

R

O

U

IIIIII..T

This volume describes how to deal with troubles with the
FLUOVIEW FV1000 system.
If any irregularity is observed, read this volume before calling
for service. If the irregularity cannot be resolved by the
described remedial action, please contact Olympus for
repair.

R

O

On This Volume

U

BLL

B

E

E

Q

Q

&

&

A

A

CONTENTS

㪈㩷㪫㫉㫆㫌㪹㫃㪼㫊㪿㫆㫆㫋㫀㫅㪾㩷㪞㫌㫀㪻㪼㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㩷㪈㪄㪈

1. Laser is not output from the extremity of the objective. 䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪈

2. Fluorescence image cannot be observed. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪈

3. Transmitted image cannot be observed. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪉

4. Image is disturbed. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃

䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪉

5. Reflected light (laser light) enters the fluorescence image. 䍃䍃䍃䍃䍃 㪈㪄㪉

6. Fluorescence image is poor.䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪉

7. Fluorescence image is dark and noisy.䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪉

8. Image is irregularly blurred or the brightness is uneven. 䍃䍃䍃

䍃䍃䍃䍃 㪈㪄㪊

9. Observed image is out of focus. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪊

10. The intensity of part of the wavelength region of the spectral

characteristic data of fluorescence is dropped.䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪊

11. Flare is observed. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃

䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪊

12. Visual fluorescent light observation is impossible. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪊

13. The light from the laser for the ASU (additional scan unit) is not

output. 䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃䍃 㪈㪄㪋

Troubleshooting Guide

1 Troubleshooting Guide

The system may be unable to manifest its full performance due to its usage as well as malfunction. In

case a problem occurs with the system please check the following list to find appropriate

countermeasures.

If the problem cannot be resolved by the described remedial action, please contact Olympus for repair.

Irregularity Cause Remedy

1. Laser is not output from
the extremity of the
objective.

The laser unit is not turned ON. Turn on the laser unit. Make sure that the

emission key is set to ON.

The laser wavelength is not selected. Check the laser wavelength to be used.

2. Fluorescence image
cannot be observed.

The manual shutter of the
fluorescence mirror unit is closed.
(Manual system only)

The reflective mirror inside the
fluorescence mirror unit is not in the
light path. (Manual system only)

The objective is not in the light path. Engage the desired objective in the light

The laser beam is too weak. Increase the laser intensity.

The properties of the combined cube
unit used for the ASU (auxiliary scan
unit) or OPD (non-confocal point
detector) do not match the selected
laser wavelength.

The confocal pinhole diameter is too
small.

The excitation Dichroic Mirror
selection does not match the
observed fluorescence wavelength
and excitation laser wavelength.

The spectral dichroic mirror and
barrier filter selections do not match
the observed fluorescence
wavelength.

Open the manual shutter.

Engage the reflective mirror in the light
path.

path. When using a manual revolving
nosepiece, be sure to stop the objective in
the click position.

Engage a DM cube unit matching the
selected laser wavelength in the light
path.

Increase the pinhole diameter.

Engage a DM optimum for the observed
fluorescence and excitation laser
wavelengths.

Engage a spectral DM and barrier filter
matching the observed fluorescence in
the light path.

The acquisition wavelength region
setting is not suitable for the
observed fluorescence wavelength.
(Spectral detection system only)

The fluorescent dyeing method and
excitation wavelength do not match
each other.

Focus is not adjusted. Adjust the focus.

The PMT voltage of the detector is
too low.

Set an acquisition wavelength region
matching the observed fluorescence.

Select a laser optimum for the fluorescent
dyeing method.

Increase the PMT voltage.

Page

III. TROUBLE Q & A

III . 1 - 1

Troubleshooting Guide

Irregularity Cause Remedy

2. Fluorescence image
cannot be observed.

The offset value is too large. Decrease the offset value.

The detector for the channel to be
detected is not selected.

3. Transmitted image cannot

be observed.

4. Image is disturbed. The system installation location is

5. Reflected light (laser light)

enters the fluorescence
image.

The transmitted light detection
channel is not selected.

The transmitted light filter for the
microscope is in the light path.

The PMT voltage of the transmitted
light detection channel is too low.

The offset value for the transmitted
light detection channel is too large.

subject to excessive vibrations.

Extraneous light such as the light of a
fluorescent lamp is detected.

The barrier filter is set erroneously or

absent.

The set acquisition wavelength is
overlapped with or too close to the
excitation laser wavelength. (Spectral
detection system only)

Select the detector.

Select the transmitted light detection
channel.

Disengage the filter from the light path.

Increase the PMT voltage.

Decrease the offset value.

Contact Olympus.

Turn the room light low before acquiring
image.

Engage a barrier filter that can cut the
excitation laser wavelength in the light
path.

㪪㪼㫃㪼㪺㫋㩷㪸㫅㩷acquisition wavelength that is
not interfered with by the laser
wavelength. (Note that, when the confocal
pinhole is large and the BS20/80
excitation DM is used, penetration of laser
light may become large.)

6. Fluorescence image is

poor.

7. Fluorescence image is

dark and noisy.

III. TROUBLE Q&A

III . 1 - 2

Page

The barrier filter that can cut the
wavelength of the laser light
irradiated from the ASU (auxiliary
scan unit) is not selected. In the case
of a spectral detection system, the
acquisition wavelength setting may
be inappropriate.

The front lens of the objective is dirty. Clean the objective front lens by wiping it

When an objective with correction
collar is in use, the correction collar is
adjusted improperly.

The cover glass thickness is
inappropriate.

The laser beam is too weak. Increase the laser intensity.

The fluorescent dyeing method and
excitation wavelength do not match
each other.

Engage a barrier filter that can cut the
laser wavelength from the ASU in the light
path. With a spectral detection system,
change the acquisition wavelength
setting.

with a piece of gauze.

Adjust the correction collar properly.

Use a cover glass with thickness of 0.17
mm.

Select a laser optimum for the fluorescent
dyeing method.

Irregularity Cause Remedy

㪎㪅㩷Fluorescence image is
dark and noisy.

The excitation Dichroic Mirror
selection does not match the
observed fluorescence wavelength
and excitation laser wavelength.

Troubleshooting Guide

Engage a DM optimum for the observed
fluorescence and excitation laser
wavelengths.

8. Image is irregularly
blurred or the brightness
is uneven.

9. Observed image is out of
focus.

The spectral dichroic mirror and
barrier filter selections do not match
the observed fluorescence
wavelength.

The acquisition wavelength region
setting is not suitable for the
observed fluorescence wavelength.
(Spectral detection system only)

The confocal pinhole diameter is too
small.

The scanning rate is too high. Decrease the scanning rate.

The HV setting is too high. Decrease the HV and increase the gain.

The width of the acquisition
wavelength region is too small.

Dyeing is too pale. Perform optimum fluorescent dyeing.

The specimen or stage is tilted. Install the specimen and stage properly.

The focus is adjusted improperly. Adjust the focus in visual observation.

Engage a spectral DM and barrier filter
matching the observed fluorescence in
the light path.

Set an acquisition wavelength region
matching the observed fluorescence.

Increase the pinhole diameter.

An alternative remedy is to decrease the
scanning rate and decrease the HV.

Increase the width of the acquisition
wavelength region.

10. The intensity of part of

the wavelength region of
the spectral
characteristic data of
fluorescence is dropped.

11. Flare is observed. The glass in use is not fluorescence-

12. Visual fluorescent light

observation is
impossible.

12. Visual fluorescent light

observation is
impossible.

The spectral characteristics of the
fluorescence are affected by those of
the excitation dichroic mirror used in
double excitation.

free glass.

The specimen is overstained. Perform optimum dyeing again or

The light path selector in the SU is
not set for the visual observation light
path.

The shutter for the mercury burner is
closed.

The mirror unit incorporating a
dichroic mirror is not present in the
turret of the illuminator.

Use an excitation DM that does not affect
the spectral characteristic data of
fluorescence.

Use fluorescence-free glass.

increase the offset value.

Select the visual observation light path.

Open the shutter for the mercury burner.

Engage a mirror unit containing DM in the
light path.

III. TROUBLE Q & A

Page

III . 1 - 3

Troubleshooting Guide

Irregularity Cause Remedy

13. The light from the laser
for the ASU (additional
scan unit) is not output.

The properties of the combined DM
cube unit for ASU do not match the
irradiated laser wavelength.

Engage a DM cube unit matching the
laser wavelength in the light path.

The combined DM cube unit for ASU
is not in the light path.

Engage the DM cube unit in the light path.

III. TROUBLE Q&A

III . 1 - 4

Page

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1.2