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Design of a Wide Field High Sensitivity Imaging System for Quantitative Analysis of CGHA Micro-Arrays.

Published online by Cambridge University Press:  02 July 2020

Damir Sudar ,
Lucas van Vliet ,
Steve Clark ,
Rick Segraves ,
Stephen Lockett ,
Donna Albertson ,
Joe Gray  and
Pinkel Daniel
Damir Sudar
Affiliation:
Life Sciences Division, E.O. Lawrence Berkeley National Laboratory, Berkeley, CA94720
Lucas van Vliet
Affiliation:
Department of Applied Physics, Delft University of Technology, Delft, The Netherlands
Steve Clark
Affiliation:
Cancer Center, University of California, San Francisco, CA94143
Rick Segraves
Affiliation:
Cancer Center, University of California, San Francisco, CA94143
Stephen Lockett
Affiliation:
Life Sciences Division, E.O. Lawrence Berkeley National Laboratory, Berkeley, CA94720
Donna Albertson
Affiliation:
Life Sciences Division, E.O. Lawrence Berkeley National Laboratory, Berkeley, CA94720
Joe Gray
Affiliation:
Cancer Center, University of California, San Francisco, CA94143
Pinkel Daniel
Affiliation:
Cancer Center, University of California, San Francisco, CA94143
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Extract

Comparative Genomic Hybridization (CGH) is a quantitative technique for determining relative copy numbers of DNA sequences in whole-genomic test samples. In conventional CGH, test DNA labelled with one fluorochrome and reference DNA labelled with a spectrally different fluorochrome are hybridized to metaphase chromosomes. by measuring the ratio of intensities of the two fluorochromes, a relative copy number of sequences in the test DNA can be calculated for each point along each chromosome. While this approach is very useful for rapid surveying of the entire test genome, the spatial and dynamic resolution are compromised by the dense packing of DNA in the metaphase state. CGHa (array-based CGH) uses spots of cloned DNA arrayed onto a microscope slide which represent the entire genome or interesting sections thereof. Spatial resolution and dynamic range are now only limited by the size of the clones used. See the abstract by Pinkel et al. for more detail.

We designed an imaging system for analyzing fluorescence signals from micro-arrays containing targets on the order of 100μm in diameter spaced at similar intervals.

Type
Light Microscopy: Recent Advances
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 811 - 812
Copyright
Copyright © Microscopy Society of America 1997

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References

1. Pinkel, D.et al., Proc. Ann. MSA Meeting 55 (1997)Google Scholar

2. Wittrup, K.D.et al., Cytometry 16:206–213 (1994)CrossRefGoogle ScholarPubMed

3. Supported by the Director, Office of Energy Research, Office of Health and Environmental Research of the U.S. Department of Energy under contract NO. DE-AC03-76SF00098.Google Scholar