Do I have to use special glass to use the FMBIO?

You will observe a higher signal to noise ratio when low fluorescence borosilicate glass is used. However, you can scan a gel with regular plates. It really depends on the particular technique and how much signal is routinely produced.

What kind of laser does the FMBIO II use?

The FMBIO II uses a powerful 50 mW solid state 532 nm YAG laser. This is important because it is much more reliable than traditional gas argon lasers and has the ability to read across a wide range of dyes from 505 to 700 nm.

What is the difference between the FMBIO and a CCD camera?

Laser based imaging systems offer significantly greater sensitivity than CCD based instruments because they are able to focus more power over a smaller spectral range. As a consequence, fluorophores are energized to a higher state thereby producing proportionally more fluorescence. The FMBIO is a laser scanner that offers much greater spacial resolution than white or UV light-based systems. The FMBIO uses 2 sets of fiber optic bundles attached to individual photomultiplier tubes. This aids in the systems increased sensitivity, speed and multi color capabilities. CCD cameras usually work over a smaller area and are limited in their multi color capabilities.

How quickly can you scan a multi-colored gel?

The FMBIO employs two photomultiplier tubes (PMTs) that collect data from two channels simultaneously. A 20 x 25 cm dual color scan is performed in three minutes.

Is the FMBIO capable of multicolor fluorescence scanning?

Yes! The FMBIO can detect fluorophores with emission wavelengths between 505 and 700 nm, providing multicolor capabilities useful in a wide variety of applications. It will scan up to 4 colors with one command using interchangeable filters and a software matrix to layer and separate the images.

Will the FMBIO read ABI dyes?

No problem. There is nothing optically special about these dyes. The FMBIO is capable of reading each of the dyes used in their four-color system. However, dye concentrations must be adjusted due to differences in detection technology. Please refer to the FMBIO dye chart for filter specifications.

Will the FMBIO detect chemiluminesence?

No, not directly. However, there is an alternative called chemifluorescence. This technique is almost identical to chemiluminesence. Simply replace the substrate included in your chemiluminesence protocol with either Vector Red?(Vector Laboratories) or Attophos (JBL). Both products are precipitable fluorescent substrates that are excitable by the FMBIO laser.

Can the FMBIO detect Green Fluorescent Protein (GFP)?

Yes, GFP is quantitative over 3 orders of magnitude on the FMBIO.

Will the FMBIO scan phosphor screens?

Absolutely not! The FMBIO is a totally non-isotopic. In fact, the FMBIO is far more sensitive than some of the combination systems that claim to detect both radioactivity and fluorescence.

Is the FMBIO compatible with SYBR dyes?

Yes. The FMBIO II is compatible with both SYBR dyes. We suggest you use the 505 emission filter to detect samples labeled with SYBR Green, and a 585 emission filter to detect samples labeled with Gold.

Can the FMBIO scan EtBr samples?

The FMBIO is 10 to 50 times more sensitive than EtBr on a transilluminator. We suggest you use the 605-nm filter.

Can I scan protein gels with the FMBIO?

You can use SYPRO Orange, SYPRO Red, or SYPRO Ruby (Molecular Probes, Eugene, Oregon). These are post-staining dyes with sensitivity higher than that of silver staining without the hassle and mess.

Can the FMBIO run sequencing gels?

The FMBIO has a scan area of 20 x 43 cm. This scan area is large enough to scan a full length sequencing gel.

Can the FMBIO scan differential display gels?

Yes, using the FMBIO requires that you run only one gel. Systems similar to the 377 require multiple gel runs -- one gel to do the analysis (destructive) and a second gel for band extraction. With the FMBIO, you run one gel, scan, remove the top plate and place a printout under the plate, line up the areas of interest and excise.

Can I recover a band(s) from my gel after detection?

One benefit of post-electrophoresis detection is that the molecules of interest are easily accessible for further manipulation. By following our simple "overlay" procedure, virtually any band is excisable.

Is the FMBIO compatible with mutation detection techniques like DGGE and SSCP?

The FMBIO is the instrument of choice for mutation detection. Fluorescence eliminates the need for tedious staining procedures, and increases throughput with multiplexing.

Why is the FMBIO considered a high throughput instrument?

Since the FMBIO is not an integrated electrophoresis / detection system, multiple gels can be run and scanned sequentially.

Can the FMBIO scan 96-, 384- and 1536-well microtiter plates based assays?

Microtiter plates of any capacity can be scanned, provided the bottom of the plate is flat, clear and the wells are black (to prevent well-to-well cross-talk).

Can I adjust the focal height of the detection optics to compensate for different media?

Yes. The proper height of the collection mirror can be quickly and easily optimized for a variety of media using the image acquisition software.

Can the FMBIO be used to scan microarrays?

Up to 48 multi-color meduim density DNA arrays can be interrogated simultaneously using the FMBIO.

What computer platform do you recommend for the FMBIO?

The FMBIO is compatible with Power Mac compatible computers and PCs. We recommend a computer with at least 64 MB RAM and at least a 500 MHz processor.

What kind of storage device do you recommend?

You can store your image files on any number of storage devices. We recommend one of the following for your lab:

• CD Writer
• Magneto Optical Disk Drive
• Zip Drive
• Jaz Drive

How large is a typical image file?

At standard resolution, a two color 20 cm x 25 cm gel is approximately 3 MB.

Can I export my images?

Yes, images can be exported as either PICT or, 8- or 16-bit TIFF formats.