The constant quest for a genome-wide analysis has routed FISH technology through a remarkable progressive path. The major developments in this journey are described below.
5.1. MULTIPLEX/MULTICOLOR FISH OR SPECTRAL KARYOTYPING A simultaneous application of carefully chosen chromosome painting probes labeled with multiple flu-orophores has made feasible to label all chromosomes in a single metaphase specimen. Both multicolor-FISH (M-FISH) and SKY serve the same purpose, but differ in their analysis procedure. In M-FISH, using selected narrow-bandpass filters separate images of each of the five fluorochromes are acquired and with a dedicated computational software that combines the unique labeling pattern of each chromosome, a distinct color index is determined for individual chromosome (43). Alternatively, in SKY, a single image is obtained through a combination of a triple-bandpass filter and Fourier transform spectrometry. Finally, spectral analysis of individual pixels of the image provides the eventual 24-color display (44). These two techniques leaped ahead of the classical metaphase banding cytogenetic analysis, in that they not only localize the marker chromosome but also trace the origins of the marker chromosome (Fig. 3). Additionally, M-FISH/SKY technology enables detection of cryptic chromosomal rearrangements as well. However, like classical cytogenetics, they suffer from the dependence on in vitro proliferation of target cells. A recent
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