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Fluorescence in situ Hybridization. Fluorescence in situ Hybridization (FISH). FISH - a process which vividly paints chromosomes or portions of chromosomes with fluorescent molecules Opening picture - Human M-phase spread using DAPI stain. Fluorescence in situ Hybridization (FISH).

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fluorescence in situ hybridization fish
Fluorescence in situ Hybridization (FISH)
  • FISH - a process which vividly paints chromosomes or portions of chromosomes with fluorescent molecules
  • Opening picture - Human M-phase spread using DAPI stain
fluorescence in situ hybridization fish1
Fluorescence in situ Hybridization (FISH)
  • Identifies chromosomal abnormalities
  • Aids in gene mapping, toxicological studies, analysis of chromosome structural aberrations, and ploidy determination
fluorescence in situ hybridization fish2
Fluorescence in situ Hybridization (FISH)
  • Used to identify the presence and location of a region of DNA or RNA within morphologically preserved chromosome preparations, fixed cells or tissue sections
fluorescence in situ hybridization fish3
Fluorescence in situ Hybridization (FISH)
  • This means you can view a segment or entire chromosome with your own eyes
  • Was often used during M phase but is now used on I phase chromosomes as well
fluorescence in situ hybridization fish4
Fluorescence in situ Hybridization (FISH)
  • Advantage: less labor-intensive method for confirming the presence of a DNA segment within an entire genome than other conventional methods like Southern blotting
fish procedure
FISH Procedure
  • Denature the chromosomes
  • Denature the probe
  • Hybridization
  • Fluorescence staining
  • Examine slides or store in the dark
fish uses
FISH Uses
  • Detection of high concentrations of base pairs
  • Eg: Mouse metaphase preparation stained with DAPI (a non-specific DNA binding dye with high affinity for A-T bonds)
fish uses2
FISH Uses
  • Centromere regions stained brighter - means they are rich in A-T bonds
  • Also used in germ cell or prenatal diagnosis of conditions such as aneuploidies
fish and telomeres
FISH and Telomeres
  • Telomeric probes define the terminal boundaries of chromosomes (5’ and 3’ ends)
  • Used in research of chromosomal rearrangements and deletions related to cell aging or other genetic abnormalities
fish and telomeres1
FISH and Telomeres
  • Special telomeric probes specific to individual chromosomes have been designed
  • Probe is based on the TTAGGG repeat present on all human telomeres
fish and telomeres3
FISH and Telomeres
  • Application in cytogenetics - can detect submicroscopic deletions and cryptic translocations of genes associated withunexplained mental retardation and miscarriages
fish and telomeres medical from telvysion dna website
FISH and Telomeres - Medical from TelVysion DNA website
  • “A translocation between chromosomes 12 and 21 with breakpoints at bands 12p13 (TEL)[telomere] and 21q22 (AML1) occurs in at least 25 percent of childhood B-cell acute lymphocytic leukemia”
fish and telomeres medical
FISH and Telomeres - Medical
  • The “LS1 TEL/AML1 ES Dual-Color Translocation Probe” is designed to detect the TEL and AML1 gene fusion
  • This fusion is undetectable by standard cytogenetics, but can be seen with FISH
fish medical
FISH - Medical
  • FISH can be used in the study of transgenic animals (eg: Polly)
  • Selective markers show if the human DNA was inserted successfully and pinpoint where the human DNA is
  • Transgenic Mouse