Tirkkonen M, Karhu R, Kallioniemi O, Isola J
Laboratory of Cancer Genetics, University of Tampere, Finland.
Cytometry. 1996 Dec 1;25(4):394-8. doi: 10.1002/(SICI)1097-0320(19961201)25:4<394::AID-CYTO12>3.0.CO;2-Q.
Comparative genomic hybridization (CGH) is based on quantitative digital image analysis of fluorescence intensities from metaphase chromosomes. High-quality CCD cameras are commonly used for image acquisition, but the minimal requirements of CCD cameras have not been determined. We first evaluated minimal camera requirements by artificially reducing spatial and dynamic resolution of images produced by a scientific-grade CCD camera (Xillix MicroImager). The results showed that reduction of dynamic resolution from 4,096 to 256 gray levels (12-bit image transformed to an 8-bit image) had negligible effect on CGH profiles and no effect on their interpretation. Similarly, CGH profiles obtained from spatially reduced images (from 1,340 x 1,035 to 670 x 517 pixels) were virtually identical to those obtained from the original image. For a practical test, we compared two 8-bit frame integrating video-rated CCD cameras (Cohu 4910 and Photometrics ImagePoint) to the Xillix Micro-Imager in a real CGH setting. Images collected from the same metaphase cells with all three cameras resulted in the identification of the same genetic changes in the samples studied. We conclude that requirements for camera resolution in CGH analysis are not stringent, and therefore that low-priced video-rated cameras capable of frame integration are sufficient for comparative genomic hybridization.
