Fluorescence in situ Hybridization - أطلس مختبرات العرب

Nearly a quarter-century has passed since the first research articles introducing in situ hybridization as a method of detecting and studying DNA sequences in chromosomes and cells appeared in the literature. Over the past 15 years, however, a revolution in light microscopy has occurred through the development of fluorescence techniques that allow unprecedented ease, precision, and accuracy in locating, identifying, and recording data on the genetic makeup of biomedical samples.

The power of in situ hybridization can be greatly extended by the simultaneous use of multiple fluorescent colors. Multicolor fluorescence in situ hybridization (FISH), in its simplest form, can be used to identify as many labeled features as there are different fluorophores used in the hybridization. By using not only single colors, but also combinations of colors, many more labeled features can be simultaneously detected in individual cells using digital imaging microscopy.

Presented in Figure 1 is a typical multicolor FISH specimen. Normal male lymphocytes were hybridized with FITC biotin-labeled Chr2l and ChrY probes and CY3 digoxigenin-labeled Chrl3 and ChrY probes. At the upper left is an image of DNA nuclei stained with DAPI, taken using a DAPI filter set. At upper right is an image of Chr21 and ChrY stained with FITC, taken using an FITC filter set. At lower left is an image of Chrl3 and ChrY stained with CY3, taken using a CY3 filter set. The image in the lower right is the joined color composite image showing all target chromosomes in color. The specimen was provided by Dr. Tim Houseal, Integrated Genetics, Framingham, Massachusetts