Development of new spectral karyotyping (SKY) techniques in cytogenetic analysis

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1. dual karyotypingThe DAPI-band image, which was digitally inverted and contrast-enhanced, was almost identical to the pattern obtained with G-banding. This method resulted in adequate band resolution that corresponded to approximately 400 bands per haploid set. The karyotypes (dual karyotypes) were ascertained by combining SKY and the DAPI-banding, in which both the DAPI-band image and the classification color image of the chromosomes are simultaneously displayed in parallel. The karyotype helped determine the origin of the rearranged chromosomal materials by using the spectrally classified color images. In addition, the breakpoints could be precisely determined with respect to chromosome bands on the G-band-like images.2. color-bandingWe developed a novel chromosome banding technique named spectral color banding (SCAN). With this technique we displayed a multicolor banding pattern that almost entirely correlated with the corresponding G-banding pattern. SCAN analysis allowed for accurate identification of chromosome-band origin on the basis of its spectral color and detection of intrachromosomal changes. Furthermore, SCAN analysis could determine the precise breakpoints of complex rearrangements and identify the chromosome-band origin of small chromosome segments. Our preliminary use of SCAN in combination with SKY suggests that this new technique is capable of accurately identifying chromosome-band origin for each of the 24 human chromosomes, and that it may be able to identify intrachromosomal changes not previously detected by G-banding or SKY.SCAN technique could become a powerful tool for cytogenetic analysis and an important step on the road to fully automated karyotyping.