Spectral karyotyping (SKY) analysis of heritable effects of radiation-induced malignant transformation

Horst Zitzelsberger; Jingly Fung; C. Janish; George McNamara; P. E. Bryant; A. C. Riches; Heinz-Ulli G. Weier

doi:10.1117/12.347579

6 May 1999 Spectral karyotyping (SKY) analysis of heritable effects of radiation-induced malignant transformation

Horst Zitzelsberger, Jingly Fung, C. Janish, George McNamara, P. E. Bryant, A. C. Riches, Heinz-Ulli G. Weier

Proceedings Volume 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI; (1999) https://doi.org/10.1117/12.347579
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

Radiocarcinogenesis is widely recognized as occupational, environmental and therapeutical hazard, but the underlying mechanisms and cellular targets have not yet been identified. We applied SKY to study chromosomal rearrangements leading to malignant transformation of irradiated thyroid epithelial cells. SKY is a recently developed technique to detect translocations involving non-homologous based on unique staining of all 24 human chromosomes by hybridization with a mixture of whole chromosome painting probes. A tuneable interferometer mounted on a fluorescence microscope in front of a CCD camera allows to record the 400 nm - 1000 nm fluorescence spectrum for each pixel in the image. After background correction, spectra recorded for each pixel are compared to reference spectra stored previously for each chromosome-specific probe. Thus, pixel spectra can be associated with specific chromosomes and displayed in 'classification' colors, which are defined so that even small translocations become readily discernible. SKY analysis was performed on several radiation-transformed cell lines. Line S48T was generated from a primary tumor of a child exposed to elevated levels of radiation following the Chernobyl nuclear accident. Subclones were generated from the human thyroid epithelial cell line (HTori-3) by exposure to gamma or alpha irradiation. SKY analysis revealed multiple translocations and, combined with G-banding, allowed the definition of targets for positional cloning of tumor related genes.

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Horst Zitzelsberger, Jingly Fung, C. Janish, George McNamara, P. E. Bryant, A. C. Riches, and Heinz-Ulli G. Weier "Spectral karyotyping (SKY) analysis of heritable effects of radiation-induced malignant transformation", Proc. SPIE 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI, (6 May 1999); https://doi.org/10.1117/12.347579

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