Olympus Fluoview User Manual

Users Manual

Fluoview

Scanning Laser Biological Microscope

Ver. 1.1

WARNING

Caution:

Before using your microscope, the items in this manual Identified by the mark shown to the left should be read carefully until completely understood to ensure safe operation.

Thank you for purchasing an Olympus Fluoview Microscope. Before using your microscope, read this manual thoroughly to make sure you obtain full performance from all of the functions provided by this system. This manual is divided into the following five sections:

For Safe Usage Introduction to Fluoview Operation Hardware Troubleshooting Q & A

OIVMPUS'

August 1996

raj

IF=°

-About This Manual

This manual explains potential risks and provide important safety information designed to ensure the safe use bf the FLUOVIEW System. Thoroughly study this manual before operating the system.

1 Safety Precautions

2 Warning Labels

CONTENTS

1-1

2-1

2-1 Warning Labels Concerning Laser Safety 2-1

2-1-1 Warning Labels 2-1 2-1-2 Aperture Label 2-3 2-1-3 Protective Housing Label 2-5

3 Precautions for Use of the System 3J^

Safety Precautions

1 Safety Precautions

Depending on the laser configuration, this laser product is classified as follows:

Ar laser configuration — CLASS n

OAyilONl

LASER RADIATION DO NOT

S[fPt

IN1D

ARGON LASER InW Mf\X

450-515nn

(LASS n LASKPRGDUCT

Kr and Ar laser configuration

CLASS in a

ANGE

LASER LIGHT

/M)IDD!fB]TBEB«RRE

KR. AR LASER SniW

MAX CW

450-570na

CLASS la

LASER

PRdXJCT

Precautions for Use

Always pay careful attention to the following points.

Avoid engaging mirror cubes into the light path in such a manner that the laser t>eam is directed outward.

This is extremely dangerous as unprotected eyes may be exposed to the laser beam.

If the cube turret of the reflected light fluorescence module, the DIC slider (optional) or the dummy slider is removed during laser output (during laser scanning), laser beam radiation will occur inside the microscope. This is extremely dangerous. Never remove the cube turret, the DIC slider, or the

dummy slider. I The cube turret, the DIC slider (optional), or the dummy slider is to be removed only by trained L personnel, and only when power has been removed from the laser system.

1-1

Safety Precautions

If the objective revolving nosepiece or the objective lens is removed during laser output (during laser scanning), the laser beam will be emitted uncontrollably. This is extremely dangerous. Never remove the objective revolving nosepiece or the objective lens.

TThe nosepiece or the objective lens is to be removed only by trained personnel, and only when [_ power has been removed from the laser system.

]

If the transmitted light lamp housing or the lamp socket is removed during laser output (during laser scanning), the laser beam light will be emitted uncontrollably. This is extremely dangerous. Never remove the reflected light lamp housing or the lamp socket.

The transmitted light lamp housing or the lamp socket is to be removed only by trained personnel,

and only when power has been removed from the laser system and from the

microscope.

• If the scan unit's (2) detection mode selector slider is detached during laser output (during laser scanning), laser beam radiation will occur inside the microscope (beam reflected from the specimen). This is very dangerous. Never remove the scan unit's detection mode selector slider.

The scan unit's (2) detection mode selector slider is to be detached only by trained personnel, |

_and only when power has been removed from the laser system. J

• Using tools to remove the protective housing, or otherwise dismantling/remodeling units, is extremely dangerous as this may lead to uncontrollable laser beam radiation inside the unit Never attempt to dismantle/remodel the system.

• Never excessively bend, strain, or step on the optical fibers. Damaged optical fibers may result in laser beam leakage. This is extremely dangerous. Should the optical fibers be damaged, immediately turn OFF the laser power supply unit and contact your Olympus representative.

• Hot air is emitted from the laser cooling fan exhaust outlet. Keep flammable materials and materials adversely affected by heat at a safe distance of the outlet.

• The power consumption of both the power unit and the laser unit is very large. Ensure that each unit obtains power from separate electrical systems.

• To prevent shock hazards, always ground the equipment.

• Always unplug the power cord before replacing fuses.

• Since the mixture (ether (70%) and alcohol (30%), etc.) used to clean the optical components of

the system is highly flammable, be careful to keep these chemicals away from open fire and potential sources of electrical sparks, such as when the main switches are moved to

"I"

(ON) or

"O"

(OFF).

1-2

Safety Precautions

Please note that the laser beam is emitted when the display's function

pane!

([Acquire]

panel) looks as shown below.

"Laser beam emitted" is shown.

w}iFjmt^axv/),^^.\jiu»:p^

Disposal of Laser Tubes

Strict adherence to applicable industrial guidelines and laws is required when disposing of laser tubes. Before disposing of laser

tubes,

contact your Olympus representative for further information.

Safety Symbols

The following symbols are found on the equipment Study the meaning of

the

symbols,

and always use

the equipment

the safest possible manner.

Symbol

Explanation Before use, carefully read the instmction manual. Improper handling could result in injury to the user and/or damage to the equipment Indicates that the main switch is ON.

Indicates that the main switch is OFF.

1-3

Warning Labels

2 Warning Labels

2-1 Warning Labels Concerning Laser Safety

2-1-1 Warning Labels

(1)

laser configuration

©AUTIOff^

LASER RADIATION

DONdTSIWE INTDB^ ARGON LASER

Inn IMX

45Ch515nn

CLASS n LASBIPRCOUCT

(2)

and

laser configuration

ANGE

LASER LIGKT MIDDIIECrBEenHIE

LASER

SaWMAX CW45O-570niD

CLASS

LASe)

PRODUCT

Attached position

BX50 configuration (Similar

case

BXWI configuration)

(Dor (2)

*^^MiiSr'

2-1

Warning Labels

IX configuration

(Dor

(2)

2-2

Warning Labels

2-1-2 Aperture Label

(Indicates where the laser beam is emitted.)

(3)

AVOID EXPOSURE

Laser radiation is emitted from this aperture

BX50 Configuration

(3) The laser beam is emitted from the

objective and the sockets on the revolving nosepiece.

(3) The laser beam Is emitted from the

condenser lens.

BXWI Configuration

(3) The laser beam Is emitted from the

objective and the sockets on the revolving nosepiece.

The laser beam is emitted from the condenser lens. 13)

2-3

Warning Labels

IX Configuration

The laser beam is emitted (3) from the objective.

(3) The laser beam is emitted from

the condenser lens.

2-4

Warning Labels

2-1-3 Protective Housing Label

(4)

CAUTION - Laser radiation when open.

DO NOT STARE INTO BEAM

t/-^f3tS®^ajit«L»c

(5)

DANGER - Laser light

when

open.

AVaO DIRECT

EYE

EXPOSURE.

CAUTION - Laser

radiation when open.

jDO NOT STARE INTO BEAM

BX50,

BXWI configuration

: (4)

Kr •»

Ar : (5)

not

remove the cube turret.

not

remove the revolving nosepiece, the objective or the DIC slider or the dummy slider.

When

the

transmitted light detector

is not

mounted (left) and when mounted (right)

(30

not

remove

the

transmitted light lamp

housing or the lamp socket.

2-5

Warning Labels

IX configuration

Ar : (4)

Kr + Ar : (5)

Do not remove the cube tuaet.

When the transmitted detection unit is not mounted (left) and when mounted (right)

Do not remove the transmitted light lamp housing or the lamp socket.

2-6

Warning Labels

Common to all configurations

When the slider cover is removed

(6) Do not remove the detection mode

selector slider.

In addition to the locations shown in this manual, a protective housing label is also attached.

For replacement of spiled brp warriihg labblSi^ pibaise cohtact

your Olympus representative. r ^^^'..'

•.:'.'

:.'r ••• '••••:;:'

•. •

2-7

3. Precautions for Use of the System

3 Precautions for Use of the System

Installation

• The FLUOVIEW system will be assembled and setup by qualified technicians. Avoid moving the system as this may adversely affect the adjustment of the optical system. In case it becomes necessary to move the system, please consult your Olympus representative. Olympus shall not be liable for improper adjustments, damages, or other problems occum'ng as a result of the system being moved by the customer or other unauthorized personnel.

• Pulling the optical fiber in case of the KrAr laser configuration will change the laser output Never modify by pulling the optical fiber.

• To prevent overheating, leave at least 30 cm of free space t>etween the exhaust opening of the

laser cooling fan and walls or other objects. Laser tube overheating will result in unstable laser oscillation or the laser oscillation may stop. Excessively high temperature inside the unit may also

result in malfunction and/or damage.

• Openings for a cooling fan for ventilation are provided on the rear panel of the control unit

computer, and the magneto-optical disk unit. To prevent the risk of overiieating and other damage, the openings should never be blocked. Leave at least 10 cm of free space between the openings and walls or other objects.

Avoid installation in places exposed to direct sunlight high humidity, high temperatures, and dust Operating environment: Ambient room temperature 10~35°C; relative humidity 30~80%.

• Avoid installation in places exposed to vibrations. Consult your Olympus representative if the location is subjected to vibrations.

Handling

• The FLUOVIEW system is a precision instrument. Handle it with care and avoid subjecting it to sudden or severe impact

• Never excessively bend, strain, or subject the laser fiber to crushing force as this will result in reduced performance.

• The piezoelectric stage and the piezoelectric nosepiece (option) employ a piezoelectric element which is fragile. Never subject to impact and ensure that undue force is never applied to these units.

Precautions for Use

• When the built-in power supply for transmitted light is tumed on for a long time, accumulated heat may cause metal to stretch and the focus to drift. To achieve highly precise image data, it is recommended to turn OFF the main switch on the microscope frame.

3-1

3. Precautions for Use of

the

System

• Standard objectives for biological use are employed for both the upright microscope system configuration (BX) and the inverted microscope system configuration (IX). Accordingly, use cover glass with a thickness of 0.17 mm or Petri dishes with a bottom thickness of 0.17 mm. It is recommended to use special cover glass for LSM water immersion objectives.

• When using the FLUOVIEW system, it is recommended to dim the light in the room to reduce the adverse effects of extraneous light.

• To ensure a stable laser beam output allow at least 10 minutes for warm-up after the laser power supply is tumed ON.

• After tuming the reflected light power supply OFF, wait at least 10 minutes before tuming ON again.

Tuming ON sooner will reduce the service life of the mercury burner.

About the Computer

and

Software

• Microsoft Windows are pre-installed on the computer for use with the FLUOVIEW system. Microsoft Windows are pre-installed on the computer for use with the FLUOVIEW system. The customer should make a back-up of the pre-installed program and store the back-up floppy disks carefully. (Olympus offers no support for back-up, missing back-up, etc.) For details on Microsoft Windows, refer to the User's Manual for Microsoft Windows.

• When the empty capacity of the hard disk becomes low, data processing will become very slow. Periodically erase unnecessary data files from the hard disk. For details on erasing data files, refer to the User's Manual for Microsoft Windows.

• When closing the software, always click the <Exit> button at the bottom of the [File I/O] panel. If you display Windows' [Task List] dialog box and click the <End Task> button to close the software, the file conditions, etc. of the last session will not be saved and subsequently not recalled next time the software is started.

• When using the Z-motor, bring the specimen into focus by operating the coarse (fine) adjustment knob on the microscope frame after clearing the check from the [Keep locked] check box on the [Z Stage] panel of the [Acquire] panel (refer to Section 4-2-3-1 in the Operation Manual). Operating the focus adjustment knobs on the microscope frame while the [Keep locked] check box remains checked may result in damage to the Z-motor.

3-2

Concerning This Section

This section provides an overview of the Fluoview System

and should be read before the system is used. After reading and understanding this section, please read the Operation section.

Contents

CAUTIONS 2 REGISTERED TRADEMARKS 2 CARE AND MAINTENANCE 3 MOVING THE FLUOVIEW 3

SECTION 1 INTRODUCTION H

1-1 Fluoview Manual Configuration 1-1

1-1-1 Users Manual Configuration 1-1

1-2 Conventions Used in This Manual 1-2

SECTION 2 SYSTEM OVERVIEW 2J.

2-1 Principle of Operation 2-1 2-2 Fluoview Features 2-2 2-3 Optical Path Diagram 2-3 2-4 System Configuration 2-4

2-4-1 System Units and Their Roles 2-4

2-5 Software Function Configuration 2-7

2-5-1 Software Panel Configuration 2-7 2-5-2 Function Panel and Display Panel 2-8 2-5-3 Drag and Drop Function Execution Icon 2-9

2-6 System Setup 2-10

2-6-1 Power Consumption 2-10 2-6-2 Tuming on the Power Supply 2-11

2-7 System Operational Procedure Overview 2-14

2-7-1 Fluorescent Light Observation Procedure 2-15 2-7-2 Transmitted Light Observation Procedure 2-16

2-8 Identifying Images From Different Methods of Observation 2-17

Cautions

(1) This software and manual may not be reproduced in part or in their entirety without the express written

permission of Olympus.

(2) The contents of this manual are subject to change without notice.

Registered Trademarks

Microsoft, Microsoft Windows and Excel for Windows are registered trademarks of Microsoft Corporation of the U.S. All other company names and product names are the trademarks or the registered trademarks of the respective companies.

Care and Maintenance

(1) When not in use, always use the accessory cover to protect your microscope from dust. (2) This microscope is a precision instrument, so do not disassemble any of the components. (3) Keep dirt, fingerprints off the lenses and filters. Remove any dirt by wiping lightly with soft gauze.

Stubborn dirt can be removed by wetting the gauze with a mixture of alcohol and ether (3:7 ratio), or benzene.

,::Ethei^

is highly <x>mbusUble^^^ cautipri when turiiihg'

* \ the main switch oh arid off.

(4) Clean the various parts by wiping with a soft cloth moistened with diluted detergent. Do not use organic

solvents because these can cause deterioration of the paint and plastic parts.

Moving the Fluoview

Avoid moving this microscope since this can adversely affect adjustment of the optical system.

Please consult with your Olympus sales representative before moving this system.

Olympus is not responsible for any problems resulting from moving this system.

Section 1 Introduction

1-1 Fluoview Manual Configuration

There are two Fluoview manuals; the Users Manual and the On-screen Manual (on-line help). The Users Manual consists of five sections. The contents of these sections are described below.

1-1-1 Users Manual Configuration

• For Safe Usage This section explains the requests, cautions and wamings related to usage of the Fluoview System.

• Introduction to Fluoview This section provides an overview of the Fluoview System.

• Operation This section explains the method of

operation,

including the input of images and different

types of image processing.

• Hardware This section provides detailed explanations of the Fluoview System hardware functions and specifications.

• Troubleshooting 0 & A This section explains the various countermeasures that can be taken in case a problem should occur.

1-1

Section 1 Introduction

1-2 Conventions Used in This Manual

The following is an explanation of

the

various conventions used in this manual.

<•

Caution,

remari< and

Symbol

one-point advisory symbols.

Explanation

Caution items are indicated by (*). Remarks to be observed and one-point advisories are

by (@).

Menu,

command button and dialogue box conventions.

Usage

Explanation

indicated

[Config] panel

<0K> button <Open File> button

The names of panels, dialogue boxes, list boxes, check boxes,

etc., are enclosed in square brackets. buttons are enclosed in triangular parentheses.

•» Mouse Operation

Convention

Explanation

Click This means to press and immediately release the mouse

button.

Double click This means to rapidly press and release the mouse button

twice.

Drag This means to hold down the mouse button, move the pointer to

the desired location and release the mouse button.

Note:

Unless othen/vise

specified,

the temns click, double click and drag are used in this manual in

relation to the left mouse button.

••• Key Operation Convention

Explanation

( Enter )

( Alt ) + fTT"

Direction keys

The names of keys are enclosed in rounded boxes(_ The plus (+) sign indicates combined key operations.

For

example,

( Alt ] + ( Fl ] means to hold down the( Alt

key and press the( Fl ] key.

( - ). CED. ( T

land,

nrikeys.

1-2

Section 1 Introduction

> Terms Unique to This System

Convention Explanation

XY obsen/ation This means using XY scan to obsen/e. (Other observation) (The same is for

XZ,

Xt, XVt and XYZt observation.)

1-3

Section 2 System Overview

The Olympus Fluoview is a confocal scanning type laser fluorescent microscope that utilizes a common focal point optical system to realize high resolution and high contrast as well as a spectacular improvement in resolution in the optical axis. This microscope provides researchers with the ability to perform automatic sectioning, three-dimensional structuring and time fluctuation observation as well as various types of image processing and analysis.

2-1 Principle of Operation

Light detector

The scanning laser microscope focuses the laser beam into a tiny spot through the objective lens and scans specimens in the X and Y directions. The scan unit detects fluorescent and transmitted light from specimens and outputs specimen

images to the image monitor.

As shown in the diagram, the confocal optical system

has a pinhole located at a position (confocal plane) which is optically conjugate with the in-focus position and which eliminates all light from outside the in-focus

position.

Consequently, these lights appears dark in

images so thick systematized specimens can be sliced optically. With ordinary optical microscopes, light from outside the in-focus position is superimposed over the image fomiing rays at the in-focus position, which results in images that are blurred. The laser beam that passes through a specimen is detected by the transmitted light detector and a transmitted light image is formed, but this is not a confocal image.

However, extremely important data can be obtained by synthesizing the transmitted light image and the confocal fluorescent light image.

Light detector

c)^

Laser

Objective lens

Specimen

2-1

Section 2 System Overview

2-2 Fluoview Features

The detector has a resolution of 12 bits, so images are extremely clear,

1 A high-resolution 1024 x 768 pixel system is used. Non-interlaced output signals are used

for clear, flickeriess images.

3. Transmitted light is detected by a photomultiplier detector, so transmitted light images are sharp and clear.

An auto-gain function eliminates any need for bothersome sensitivity adjustment

5. Two image modes, 2-channel fluorescent images or fluorescent image + transmitted light image,

can be selected with one touch.

6. The scan unit is corrected up to the infrared range to assure compatibility with a variety of lasers. Use is also possible with both erect image and inverted image microscopes.

7. The confocal aperture is a 5-position turret, so the correct confocal aperture for each objective lens can be selected with one touch.

2-2

Section 2 System Overview

2-3 Optical Path Diagram

CHI CH2

FL TR

Laser

CH2

CHI CH2

TR FL

CH2

PM2

• —

2-3

+ 56 hidden pages

Olympus Fluoview User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual