Evaluation of camera requirements for comparative genomic hybridization

✍ Scribed by Mika Tirkkonen; Ritva Karhu; Olli Kallioniemi; Jorma Isola

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 438 KB
• Volume: 25
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19961201)25:4<394::aid-cyto12>3.0.co;2-q

✦ Synopsis

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 artiticially reducing spatial and dynamic resolution of images produced by a scientiac-grade CCD camera (xillir MicroImager). The results showed that reduction of dynamic resolution from 4,096 to 256 gray levels (12-bit image transformed to an &bit image) had negligible effect on CGH profiles and no effect on their interpretation. Similarly, CGH profiles obtained from spatially reduced images (Srom 1,340 X 1,035 to 670 X 517 pixels) were virtually identical to those obtained from the original image.

MATERIALS AND METHODS

DNA Samples and CGH Hybridization

High molecular weight DNA was isolated from the MCF-7 breast cancer cell line and from the peripheral blood of karyotypically normal male and female donors.