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FISH ( FLUORESCENCE IN SITU HYBRIDISATION)

FISH ( FLUORESCENCE IN SITU HYBRIDISATION). BY: VASWATI DAS R.N:8 M.SC 2 ND SEM. FISH. It is a cytogenetic technique. Uses fluorescent molecules to “paint” genes or chromosomes.

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FISH ( FLUORESCENCE IN SITU HYBRIDISATION)

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  1. FISH(FLUORESCENCE IN SITU HYBRIDISATION) BY: VASWATI DAS R.N:8 M.SC 2ND SEM

  2. FISH It is a cytogenetic technique. Uses fluorescent molecules to “paint” genes or chromosomes. Identifies chromosomal abnormalities.Aids in gene mapping,toxicologicalstudies,analysis of chromosome structural aberrations,andploidy determination

  3. Used to identify the presence and location of a region of DNA or RNA within morphologically preserved chromosome preparations. We can view a segment or entire chromosome with our own eyes. Often used during M phase,but is now used on I phase chromosome as well.

  4. PROCEDURE Denature the chromosome. Denature the probe. Hybridization. Fluorescence staining. Examine slides or store in the dark.

  5. HYBRIDIZATION PROBE It is a fragment of DNA or RNA of variable length. Used in DNA or RNA samples to detect the presence of nucleotide sequences(the DNA target) that are complementary to the sequence in the probe.

  6. TYPES OF FISH Stellaris FISH Fiber FISH Q-FISH Flow FISH

  7. ADVANTAGES Rapid and sensitive. Less labour intensive method than other conventional methods like Southern blotting. Lots of cells can be analysed. No cell culture needed. Result may be easier to interpret than interpreting of complex karyotype.

  8. DISADVANTAGES Need specialised camera and image capture system. Limited no of commercial probes available. It will provide information about the probe being tested,other aberrations will not be detected.

  9. MEDICAL APPLICATIONS Chromosomal deletion causing William Syndrome is so small that it can only be observed by using FISH. Other diseases diagnosed are Prader-Willisyndrome,Anglemansyndrome,Chronicmyelogenousleukemia,Down syndrome. It can be used to form a diagnosis,to evaluate prognosis or to evaluate remission of a disease’such as cancer.

  10. USES Deletion of high concentration of base pairs.Eg:Mouse metaphase preparation stained with DAPI. Used in clinical studies. Used to compare the genomes of two biological species,to deduce evolutionary relatioships. Used in the field of microbial ecology,to identify microorganisms.

  11. A metaphase cell positive for the bcr/abl rearrangement (associated with chronic myelogenous leukemia) using FISH. The chromosomes can be seen in blue. The chromosome that is labeled with green and red spots (upper left) is the one where the wrong rearrangement is present.

  12. Thank you

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