FUJIFILM FLA-5OOO Fluorescent Image Analyzer Instruction Manual

Science Imaging Systems
Very Wide Array
Fluorescent/Radioisotope
Science Imaging
System
FLA-5OOO
Instruction Manual

FLA-5OOO Fluorescent Image Analyzer

Advanced fluorescent and radioisotope detection for next-generation imaging

Versatile

More imaging methodologies in a single system than ever before
The future of science imaging continues to unfold with Fujifilm’s next- generation FLA5000 imaging system.
More versatile than any predecessor, the FLA-5000 provides four separate imagingmethodologies: Fluorescent, Radioisotopic, Chemiluminescent and Gel Documentation.
With a large 40 x 46cm sampling area, scanning pixels as low as 25 microns and modular add-ons for key imaging components, the FLA-5000 takes imaging versatility to a new level.

Modular
Performance for today with adaptability for tomorrow
The FLA-5000’s modular design offers a unique opportunity to expand the system’s capabilities and performance as research methodologies evolve. A fourth laser easily may be added to the system’s three standard lasers to increase productivity or to excite new fluorescent dyes as they become available. A second photomultiplier tube (PMT) coupled with an additional optical filter may be added to accommodate concurrent or dual fluorescent detection.
The system’s modular design accommodates today’s imaging needs and tomorrow’s imaging expectations.

Sensible

Grows when you grow, but not before
Today’s researchers demand more from their imaging systems than ever before. They want their investments to do more, last longer and adapt to the future. They want more imaging methodologies in a single system with greater versatility and higher resolutions. They want to expand a system’s capabilities, not replace them. They want a system to make sense.
The FLA-5000 Science Imaging System.
It makes sense.

Largest Scanning Area
A large scanning size (up to 40 x 46cm with selectable scanning area) is especially suitable for large size 2D protein electrophoresis gel analysis.
The FLA-5000 accommodates the following Imaging Plates (IPs): 1x (3543 MS IP), 2x (2340, 2325 MS IP) and 2x (2040, 2025 MS, SR, ND, TR IP).
25-micron Scanning Pixels Scanning pixels are user-selectable at 25, 50, 100 or 200-micron pixels depending on specific methodology requirements.
Multiple Lasers
Multiple lasers add imaging applications.
The FLA-5000 can include three internal BGR lasers as standard: SHG blue laser (473nm), SHG green laser (532nm) and LD red laser (635nm). As an option one additional laser can be added in the future allowing selection of up to four lasers. The optional fourth laser can be connected to either an internal or external port.

Multi-format Detection
The versatility of multi-format detection allows many sampling opportunities with a single sampling tool.
Fluorescence detection by laser scanning:  Repetitive scanning of a sample withdifferent lasers and filters is controlled by Image Reader software. The addition of an optional second PMT and filter allows the simultaneous detection of two different fluorescent dyes by two different exciting lasers in a single scan.
Radioisotope by IP method:IP-S (standard mode for RI detection) generates logarithmic converted values in PSL units along with linear TIFF. The selection of the types of files generated by Image Reader software can be done. IP-V (variable mode) can change the high voltage of the PMT, which enables X-ray diffraction study, nondestructive testing and other IP detection studies.
Chemiluminescence by direct detecting:
Chemiluminescence can be detected.
Gel Documentation function by Epi-illumination with fluorescent screen:Applicable for transparent gels with silver stain, CBB stain, NBT stain and others.
Stages
The Fluor Stage, Multi Stage and IP Stage allow multiple detection opportunities, including: radioisotopic images, agarose gel, polyacrylamide gel, differential display with glass sheets, membrane and others.
Fluor Stage:The Fluor Stage includes a 40 x 46cm glass platen with an optional gel stopper and is used for fluorescent detection, gel documentation function and chemiluminescent detection. With the addition of the optional second  PMT concurrent detection of dual fluorescence is possible.

Features

Multi Stage:The Multi Stage is used for detecting fluorescence in glass plate or in up to six microtiter plates with the optional microtiter plate holder.
IP Stage:To capture radioisotopic images the magnetic IP Stage holds the IP with a soft ferrite backing layer of any size up to 40 x 46cm. Recommended IP is BAS-MS type.
Easy to Use
Easier to Maintain
Fujifilm software: The FLA-5000 imaging system is fully supported by Fujifilm’s existing software packages, including Science Lab (Image Gauge and L Process), Array Gauge and Multi Gauge.
Removable stages:The IP, Fluor and Multi stages are easily removed from and inserted into the top of the imaging unit for convenient detection of samples.
Since the Fluor Stage is also waterproof, the gel can be han- dled with excess water on the stage. Removable filter cartridges:The filter cartridges are removable and easily changed to accommodate specific detection criteria.

Imaging

Fluorescence
Fluorescent image with Fluor Stage and Multi Stage
Three excitation lasers are used to create images of fluorescently labeled or fluorescently stained samples.

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DNA stained with SYBR® Green I after electrophoresis with agarose gel (473nm excitation laser with LPB filter)| Protein stained with SYPRO® Ruby after 2D electrophoresis (473nm excitation laser with LPG filter)

Gel Documentation
Gel Documentation image with Fluor Stage
The gel documentation function is used for silver stained or CBB stained gel imaging. A specific fluorescent screen is placed over the stained gel on a Fluor Stage. Silver stained or CBB stained bands absorb the excitation laser and the emission light of the fluorescent screen and generate a posi/nega converted image. The PMT is set to 250v. The resulting reversed black and white images are automatically acquired by Image Reader software.

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Silver stained gel (473nm excitation laser with LPB filter)| CBB stained gel (532nm excitation laser with LPG filter)

Optical Path

• Laser Light Source – Three internal lasers (blue, green and red) are standard with an additional internal bay available for an optional fourth laser.
• Stage – Laser light is reflected by a mirror into the opticl head, which moves under the 40 x 46cm stage. The optical head directs the laser light onto the sample.
• Detection – Light emitted from the sample travels through the filter to the PMT for channel 1.
  The addition of an optional PMT for channel 2 enables simultaneous dual wavelength excitation and dual wavelength detection with optional filters for FITC/Cy5 and Cy3/Cy5.

Radioisotope
Radioisotope image with IP Stage
A maximum image size of 46 x 40cm can be detected on the FLA-5000. Depending on the sampling requirement the system will accommodate a single BAS-MS3543 IP or two concurrent BAS-2340/2040/2325/2025 IPs.

Macroarray image of ResGen™ membrane
Data courtesy of Dr. Takahashi, Aichi Cancer Research Center

Chemiluminescence
Chemiluminescent image with Multi Stage or Fluor Stage
The optical scanning head directly detects chemiluminescence. When pixel size is set to 200 microns, a 5 x 10 cm sample can be scanned in about two minutes.

DNA (pBR328) image with CDP-Star (The band at the upper right (8th slot) corresponds to 500fg.)| The same sample is detected by the LAS-1000plus system with a five-minute exposure.
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Applicable Software

The FLA-5000 utilizes Fujifilm’s familiar, user-friendly software packages researchers have depended on for years to simplify their imaging, analysis and reporting functions.

• Science Lab (Image Gauge and L Process)
• Array Gauge
• Multi Gauge – Designed specifically for the FLA-5000 system, Multi Gauge builds on Fujifilm’s reputation for developing fast, easy-to-use software interfaces to simplify
  highly complex research functions.

Image overlapping by Multi Gauge

Multi Gauge software is able to analyze multi-channel fluorescence data from the FLA-5000 imaging system. The images can be overlapped, or viewed in parallel, for profile analysis.

Specifications and Applications

Operating System
Windows® and MacOS
Nuclides
14C,32P,33P,35S,125I,3H, Neutron, etc.
Dynamic Range
Five orders of magnitude
Bit Depth
16 bit/8bit
Recommended Use
The IP, fluorescence, gel documentation and chemiluminescence detection ability allows multipurpose use.
Filters
Four filters may be placed on a single filter tray and may be easily changed.
Available filters are indicated below in Accessories.
Dimensions (mm)
900 (W) x 800 (D) x 400 (H) mm
Weight
ca. 110kg

Fluorescent dyes with suitable excitation wavelength (laser) and filter

473 nm

532nm

635nm

Standard filters:
LPB (Y510), LPG (0575), LPR (R665)
Optional filters:
BPB1 (530DF20), BPG1 (570DF20), DBR1 (530DF20, R665), DGR1 (570DF20, R665)

Accessories

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Filter Tray
Filters
Filter for IP detection
Y510 for blue laser
O575 for green laser
R665 for red laser
530DF20 for FITC
570DF20 for Cy3™
530DF20/R665 set for FITC/Cy5™
570DF20/R665 set for Cy3™ /Cy5™
Pictrography 3500/4000| Fluorescent screening for digitizing function
Additional PMT (option) for concurrent detection of dual fluorescence

From left to right:
FLUOR Stage
(with optional gel stopper)
A glass platen of 40 x 46cm can be used for fluorescence detection, digitizing function and chemiluminescence detection.
IP Stage
A magnetic stage holds the IP with a soft ferrite layer of any size up 40 x 46cm.
MULTI Stage
(with optional microtiter plate holder which can hold up to six microtiter plates)
Large size polyacrylamide gel plate can be measured with the glass.

Fuji Photo Film Co., Ltd. 26-30, Nishiazabu 2-Chome, Minato-ku, Tokyo 106-8620, Japan, Tel: +81-3-3406-2201, Fax: +81-3-3406-2158 • http://home.fujifilm.com/products/science/index.html • E-mail: sginfo@tokyo.fujifilm.co.jp
Fujifilm Medical Systems U.S.A., Inc. 419 West Avenue, Stamford, CT 06902, U.S.A. Tel: +1-203-324-2000 ext. 6112 (1-800-431-1850 ext. 6112 in the U.S.) Fax: +1-203-351-4713 • http://www.fujimed.com • E-mail: SSG@fujimed.com
Fuji Photo Film (Europe) GmbH, Heesenstr. 31, 40549 Düsseldorf, Germany, Tel: +49-211-5089-174 Fax: +49-211-5089-139 • http://www.fujifilm.de • E-mail: mkaling@fujifilmeurope.de
Specifications and system configuration subject to change for improvement without notice. All other product names mentioned herein are the trademarks of their respective owners.

References

• 富士フイルム [日本]
• fujimed.com

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