Figure 3. Schematic representation of spectral karyotyping hybridization experimental set up.
(A) Flow sorting is used to isolate individual chromosomes. (B) Each chromosome is then labeled with a unique combination of fluorescent dyes or haptens. (C) The hybridization cocktail (a combination of these differentially labeled chromosome painting probes and Cot-I DNA to suppress the hybridization of repeat sequences) is hybridized to tumor metaphase preparations. (D) After washes and detection of the hapten labeled probes, the metaphases are imaged using a fluorescent microscope. (E) A spectrophorometer in the imaging system generates a spectral signature for each pixel in the image. (F) Each pixel is assigned a classification color based on its spectral signature. Each normal chromosome is therefore a single color.
