Olympus Fluoview FV1000 User manual

User’s Manual
FLUOVIEW
FV1000
CONFOCAL LASER SCANNING
BIOLOGICAL MICROSCOPE
[HARDWARE]
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. This user’s manual is composed of three volumes including “SYSTEM
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.
AX7282

CAUTION
㪈
Page
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.

MANUAL CONFIGURATION
㪉
Page
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

NOTATIONS IN THIS MANUAL
㪊
Page
NOTATIONS IN THIS MANUAL
This manual complies with the following notations.
Notation of Caution, Notes and Tips
Notation Description
Caution to prevent injuries to the user or damage to the
product (including surrounding objects).
NOTE
Note for the user.
TIP
Hint or one-point advice for user reference.

This volume describes the overview of the FLUOVIEW
FV1000 system.
Please read this volume so that you can understand the
system before use.
On This Volume
I
I. .S SY YS ST TE EM MO OV VE ER RV VI IE EW W

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
I. System Overview
I. 1-1
Page
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.
Confocal
aperture
Light detector
Objective
Specimen
Light detector
Laser

System Overview / Features of the FV1000
I. System Overview
I. 1-2 Page
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.

System Overview / Optical Path Diagram
I. System Overview
I. 1-3
Page
1-3 Optical Path Diagram
Mercury light Supply
Laser beam
Galvano mirror
IR
UV
VIS
Galvano mirror
DM
DM
DM
DM
Barrier filter
Barrier filter
Slit
Slit
Ch1
Ch2
Ch3
Grating
Grating
Pinhole
Ch4

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
Detector
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 4th 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
Review
Software
Filter Set
for SPD MBGR
Filter Set
for FD
MBGR

System Overview / System Configuration
I. System Overview
I. 1-5
Page
1-4-2 System Appearance and Functions
The applicable microscopes are the BX61/62TRF, BX61WIF and IX81F.
Top View
㪝㪠㪩
Laser Power Supply
Microscope
Epi Fibe
r
Illumination Unit
Mouse
Keyboard
PC
A
VT㩷
UCB
Microscope
COMB
RFL TD
LCD LCD
PSU
Power supply
Power supply
Scan Unit
Heart of lase
r
scanning
microscope, composed
of scanner and light detector.
Epi Fiber Illumination Unit
Illumination unit based on
mercury burner, connected to
the microscope through a fiber. Laser power supply
Power supply for the
Ar, HeNe green
and HeNe red lasers.
20’ Flat Panel Display
Monitors for displaying
the laser scanning image
and control panel, etc.
Power supply unit
A
lso controls the FV1000 scan
unit, laser combiner, etc.
System Controller
Used to control
the FV1000, file
its images, etc.
Laser combiner
Unit for combining the laser light
of the Ar, HeNe green and
HeNe lasers into a single fiber.
Anti-vibration table
Control box
Controls
the microscope.
Mercury power suppl
y
Power supply for reflected
light illumination.
Transmitted light detector
(incl. transmitted light illumination unit)
Unit for obtaining the transmitted
image, connected to the
microscope through a fiber.
Microscope
Designed for fluorescence
observations.
Keyboard
Front View

I
II I. .P PR RE EP PA AR RA AT TI IO ON N
F
Fo or rO OP PE ER RA AT TI IO ON N
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.
On This Volume

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