30 Microdissection of Chromosomes and Reverse FISH
401
7. Apply 20 III of an antifade solution (e.g. Vectashield) and cover with a
24 x 60 mm coverslip.
Slides are ready for microscopic evaluation.
Reverse FISH
In reverse FISH experiments the labeled DNA-library of an aberrant chromosome, or parts of it, is hybridized to normal metaphase spreads. In
order to ensure a precise breakpoint definition, metaphase plates should
be well spread and chromosomes sufficiently long. A well defined DAPI
banding is obtained when the slides are aged for 5-7 days at room temperature prior to hybridization.
When clinical cases are analyzed, precautions should be taken to ensure maximum reliability of the results. Especially when aberrant chromoFig. 2. Reverse FISH of a marker chromosome is shown. The DNA library is hybridized
back to the patient's metaphase spreads. Interpretation of these results leads to the diagnosis of a der(2I)(llqterllqI4,2::2Ipllcenqter). Localization of the involved chromosomal
regions into the formation of the derivative chromosome is indicated by square brackets
next to the chromosomes ideograms
401
7. Apply 20 III of an antifade solution (e.g. Vectashield) and cover with a
24 x 60 mm coverslip.
Slides are ready for microscopic evaluation.
Reverse FISH
In reverse FISH experiments the labeled DNA-library of an aberrant chromosome, or parts of it, is hybridized to normal metaphase spreads. In
order to ensure a precise breakpoint definition, metaphase plates should
be well spread and chromosomes sufficiently long. A well defined DAPI
banding is obtained when the slides are aged for 5-7 days at room temperature prior to hybridization.
When clinical cases are analyzed, precautions should be taken to ensure maximum reliability of the results. Especially when aberrant chromoFig. 2. Reverse FISH of a marker chromosome is shown. The DNA library is hybridized
back to the patient's metaphase spreads. Interpretation of these results leads to the diagnosis of a der(2I)(llqterllqI4,2::2Ipllcenqter). Localization of the involved chromosomal
regions into the formation of the derivative chromosome is indicated by square brackets
next to the chromosomes ideograms