mFISH | mFISH
Key Words: fluorophore, fluorescence, FISH, CGH
Definition:mFISH uses many fluorescent dyes to simultaneously characterize chromosomes in one hybridization step
TECHNOLOGY:
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mFISH uses multiple fluorescent probes to achieve a distinctive color signature for each chromosome. 'Combinatorial' labeling, which involves labeling each chromosome with a combination of a limited number of dyes, is used to overcome the limitations of cleanly resolving many fluorphores. Separation of excitation and emission spectra is through the use of appropriate filter sets. mFISH requires highly sensitive probe detection.
Colors created in mFISH may not be distinguishable by the human eye. Pseudo-color allows more easily identifiable colors to be assigned to each probe. Chromosomal rearrangements can be observed as color changes within an affected chromosome. The technique allows the rapid identification of simple and complex chromosomal alterations in metaphase spreads. Fluorescence lifetime measurements, and the use of long lifetime probes, provide further possibilities in providing contrast in mFISH studies.
APPLICATIONS:
Applicable to biomedical research and cytogenics studies, mFISH is used to identify non-random structural chromosome rearrangements not detectable by other methods. These rearrangements may be associated with disease (especially cancer) initiation and progression.
MICROSCOPE CONFIGURATION:
mFISH requires a research level fluorescence upright microscope with a motorized fluorescence turret. Image analysis software is required for pseudo-color assignment and interpretation. High-transmission objectives, free of color and spherical aberrations, are essential for imaging minute areas on chromosomes.
RECOMMENDED SYSTEM:
The Eclipse 90i is ideally suited to the capture of low light fluorescence data and enables automated image capture. A variety of commercially available cytogenetic data analysis software packages are available to facilitate image interpretation.
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