Olympus Fluoview FV1000 Guide d'installation

Confocal Laser Scanning

Biological Microscope

FV1000

FLUOVIEW

FLUOVIEW—Always Evolving

FLUOVIEW–—From

Olympus is Open

FLUOVIEW—More Advanced than Ever

The Olympus FLUOVIEW FV1000 confocal laser scanning microscope delivers

efficient and reliable performance together with the high resolution required for

multi-dimensional observation of cell and tissue morphology, and precise

molecular localization. The FV1000 incorporates the industry’s first dedicated

laser light stimulation scanner to achieve simultaneous targeted laser stimulation

and imaging for real-time visualization of rapid cell responses. The FV1000 also

measures diffusion coefficients of intracellular molecules, quantifying molecular

kinetics. Quite simply, the FLUOVIEW FV1000 represents a new plateau, bringing

“imaging to analysis.”

Olympus continues to drive forward the development of FLUOVIEW

microscopes, using input from researchers to meet their evolving demands and

bringing “imaging to analysis.”

Quality Performance with Innovative Design

FV10i

Imaging to Analysis

ing up New Worlds

From Imaging to Analysis

FV1000

Advanced Deeper Imaging with High Resolution

FV1000MPE

Advanced FLUOVIEW Systems Enhance the Power of

Your Research

Superb Optical Systems Set the Standard

for Accuracy and Sensitivity.

Two types of detectors deliver enhanced accuracy and sensitivity,

and are paired with a new objective with low chromatic aberration,

to deliver even better precision for colocalization analysis.

These optical advances boost the overall system capabilities and raise

performance to a new level.

Imaging, Stimulation and Measurement—

Advanced Analytical Methods for Quantification.

Now equipped to measure the diffusion coefficients of intracellular

molecules, for quantification of the dynamic interactions of molecules

inside live cell.

FLUOVIEW opens up new worlds of measurement.

Evolving Systems Meet the Demands of Your Application.

Upgradeable system with optional hardware and software to meet

the demands of your research.

Laser combiner/Fiber

Diode Laser

Greater stability, longer service life and

lower operating cost are achieved using

diode lasers.

Laser Feedback Control

Scanner unit is equipped with laser power

monitor for feedback control enhancing

stable laser output.

Laser Compatibility

Diode laser :

405 nm, 440 nm, 473 nm, 559 nm, 635 nm

Gas laser :

Multi-line Ar laser (458 nm, 488 nm, 515 nm)

HeNe(G) laser (543 nm)

Broadband Fiber

Broadband fiber connection for 405–635 nm

lasers, to achieve an ideal point light source

with minimal color shift and position shift

between images.

LD405

LD473

LD559

LD635

AOTF

Grating

Barrier filter

Confocal pinhole

Grating

AOTF

Laser combiner

Main scanner

Broadband fiber *

Broadband fiber

Excellent Precision, Sensitivity and Stability.

FLUOVIEW Enables Precise, Bright Imaging with Minimum Phototox

Two versions available.

•Single fiber-type combiner is used for

main scanner FV1000 with up to six lasers,

ranging from 405 to 635 nm.

•Dual fiber-type combiner is used for laser

light stimulation with main and SIM

scanner FV1000.

Laser Combiner

Scanners/Detection

High Sensitivity Detection System

High-sensitivity and high S/N ratio optical

performance is achieved through the

integration of a pupil projection lens, use

of a high sensitivity photomultiplier tube

and an analog processing circuit with

minimal noise. Enables high S/N ratio

image acquisition with minimal laser power

to reduce phototoxicity.

Up to Four PMT Channels

Three integrated confocal PMT detectors,

and optional module with fourth confocal

PMT expandable up to four PMT channels.

Two Versions of Light Detection System

• Spectral detection for high-precision

spectroscopy with 2 nm resolution.

• Filter detection equipped with high

quality filter wheels.

Spectral Scanning Unit Filter Scanning Unit

Technology / Hardware

IX81

PMT

Galvanometer

scanning mirrors

Galvanometer

scanning mirrors

UIS2 objectives

Specimen

Pupil

projection

lens

SIM Scanner *

Microscope

icity.

Optical System

Motorized Microscopes

Compatible with Olympus IX81 inverted

microscope, BX61WI focusing nosepiece

and fixed-stage upright microscope, and

BX61 upright microscope.

Samples and Specimens

Supports a Wide Range of Samples and

Specimens

Tissue culture dishes, slide chambers,

microplates and glass slides can be used

with live cells and fixed specimens.

BX61

UIS2 Objectives

Olympus UIS2 objectives offer world-

leading, infinity-corrected optics that

deliver unsurpassed optical performance

over a wide range of wavelengths.

High S/N Ratio Objectives with

Suppressed Autofluorescence

Olympus offers a line of high numerical

aperture objectives with improved

fluorescence S/N ratio, including

objectives with exceptional correction for

chromatic aberration, oil- and water-

immersion objectives, and total internal

reflection fluorescence (TIRF) objectives.

* Option

Spectral Based Detection

Flexibility and High Sensitivity

Spectral detection using gratings

for 2 nm wavelength resolution

and image acquisition matched to

fluorescence wavelength peaks.

User adjustable bandwidth of

emission spectrum for acquiring

bright images with minimal cross-

talk.

Precise Spectral Imaging

The spectral detection unit uses a grating method that offers

linear dispersion compared with prism dispersion. The unit

provides 2 nm wavelength resolution to high-sensitivity

photomultiplier tube detectors. Fluorescence separation can be

achieved through unmixing, even when cross-talk is generated

by multiple fluorescent dyes with similar peaks.

496 500 504 508 512 516 520 524

Wavelength

Intensity

528 532 536 540 544 548 552

400

600

800

1,000

1,200

1,400

1,600

1,800

2,000

2,200

2,400

2,600

EGFP (dendrite) — EYFP (synapse)

XYλ

Wavelength detection range: 495 nm–561 nm in

2 nm steps

Excitation wavelength: 488 nm

Courtesy of: Dr. Shigeo Okabe

Department of Anatomy and Cell Biology,

Tokyo Medical and Dental University

EGFP–EYFP Fluorescence Separation

EYFP

EGFP

480

Conventional mirror unit High-performance mirror unit

500 520 540 560 580

Wavelength (nm)

600 620 640 660 680 700

Transmittance (%)

100

DM488/543/633 Comparison

Two Versions of Light Detection System that Set New Standards

for Optical Performance.

Filter Based Detection

Enhanced Sensitivity

Three-channel scan unit with detection system featuring hard

coated filter base. High-transmittance and high S/N ratio optical

performance is achieved through integration of a pupil projection

lens within the optics, the use of a high sensitivity photomultiplier

and an analog processing circuit with minimal noise.

High-Performance Filters Deliver Outstanding Separation

Special coatings deliver exceptionally sharp transitions to a

degree never achieved before, for acquisition of brighter

fluorescence images.

Technology / Hardware

SIM (Simultaneous) Scanner Unit

Combines the main scanner with a dedicated laser light

stimulation scanner for investigating the trafficking of fluorescent-

labeled molecules and marking of specific live cells.

Simultaneous Laser Light Stimulation and Imaging

Performs simultaneous laser light stimulation and imaging to

acquire images of immediate cell responses to

stimulation in photobleaching experiments.

Modifiable Stimulation Area During Imaging

The stimulation area can be moved to a different position on the

cell during imaging, providing a powerful tool for photoactivation

and photoconversion experiments.

Wide Choice of Bleaching Modes

Various scan modes can be used for both the observation area

and stimulation area. Enables free-form bleaching of designated

points, lines, free-lines, rectangles and circles.

SIM Scanner Unit for Simultaneous Laser Light Stimulation and

Imaging.

Branching of laser in

laser combiner.

Lasers are used for both imaging

and laser light stimulation.

LD405

LD635

AOTF

LD473

LD559

Multi-Purpose Laser Combiner

All lasers can be used for both Imaging and laser light stimulation.

LD405 / LD635 / AOTF / AOTF / LD473 / LD559

Laser Sharing with Main Scanner

Dual fiber laser combiner provides laser sharing between the SIM

scanner and main scanner, eliminating the need to add a

separate laser for stimulation.

Unique "Tornado" Scanning for Efficient Bleaching

Conventional raster scanning does not always complete

photobleaching quickly. Tornado scanning greatly improves

bleaching efficiency by significantly reducing unnecessary

scanning.

*Tornado scanning only available for SIM scanner.

Tornado scanning

Superfluous scanning areas.

ROI (region of interest)

scanning

ROI (region of interest) scanning.

Tornado scanning.

Cell membrane stained with DIO, and subjected to both

conventional ROI and tornado scanning.

Low Chromatic Aberration Objective

Best Reliability for Colocalization Analysis

A new high NA oil-immersion objective minimizes chromatic

aberration in the 405–650 nm region for enhanced imaging

performance and image resolution at 405 nm. Delivers a high

degree of correction for both lateral and axial chromatic

aberration, for acquisition of 2D and 3D images with excellent

and reliable accuracy, and improved colocalization analysis. The

objective also compensates for chromatic aberration in the near

infrared up to 850 nm.

New Objective with Low Chromatic Aberration Delivers

World-Leading Imaging Performance.

NEW

Low Chromatic PLAPON60xOSC

Aberration Objective Magnification: 60x

NA: 1.4 (oil immersion)

W.D.: 0.12 mm

Chromatic aberration compensation range: 405–650 nm

Optical data provided for each objective.

3D image

Tubulin in Ptk2 cells labeled with

two colors (405 nm, 635 nm) and

compared.

Improved Flatness and Resolution at 405 nm

Better flatness reduces the number of images for tiling.

450400

-0.5

0.0

Focal plane (µm)

0.5

Lateral Chromatic Aberration

Wavelen

th (nm)

500 550 600 650

PLAPON 60xOSC

UPLSAPO60xO

Chromatic Aberration Comparison for PLAPON 60xOSC and

UPLSAPO 60xO

Performance Comparison of PLAPON 60xOSC and UPLSAPO 60xO

PLAPON60xOSC

Axial chromatic

aberration (Z direction)

Compared for PSF fluorescent

beads (405 nm, 633 nm).

Lateral chromatic

aberration (X-Y direction)

Compared for PSF fluorescent

beads (405 nm, 488 nm, 633

nm).

*Chromatic aberration values are design values and are not guaranteed values.

Lateral and Axial Chromatic Aberration

Small Degree of

Chromatic Aberration

Large Degree of

Chromatic Aberration

Axial chromatic

aberration

(Z direction). Lateral chromatic

aberration (X-Y

direction at FN6).

Approx.

0.5 µm

Approx.

0 µm

Approx. 0.1 µm Approx. 0.2 µm

Objective

PLAPON60xOSC UPLSAPO60xO

Flatness Comparison Image at 1x Zoom

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