Semiautomated DNA probe mapping using digital imaging microscopy: II. System performance

Masaru Sakamoto, Daniel Pinkel, Laura Mascio, Damir Sudar, Donald Peters, Wen‐Lin ‐L Kuo, Kazuhiro Yamakawa, Yusuke Nakamura, Harry Drabkin, Zeljiko Jericevic, Louis Smith, Joe W. Gray

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

This paper describes an evaluation of a semiautomated, multicolor image‐analysis system to map cloned probes along metaphase chromosomes. Mapping with this system consists of fluorescence in situ hybridization (FISH) for probe localization, automatic acquisition of multicolor images showing total chromosomal DNA and probe location(s), and automatic determination of the fractional locations of the probes along the chromosomes relative to the short arm telomere (FLpter). The system was evaluated by mapping ten phage and ten cosmid probes previously mapped to chromosome 3 with other procedures. The standard deviations of FLpter measurements averaged 3.4 Mb and 2.6 Mb for phage and cosmid probes, respectively. With this variation, the order of two probes mapped in separate hybridizations could be determined with 95% confidence when their separation was greater than 2.5 Mb. In all cases, the probe locations and order were consistent with previous mapping data. FLpter values were converted to band locations using measurements of the band locations made using digital imaging microscopy. This proved superior to conversions made using ISCN ideograms. © 1995 Wiley‐Liss, Inc.

Original languageEnglish (US)
Pages (from-to)60-69
Number of pages10
JournalCytometry
Volume19
Issue number1
DOIs
StatePublished - Jan 1 1995

    Fingerprint

Keywords

  • FISH
  • FLpter
  • digital microscopy
  • image analysis
  • mapping

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

Cite this

Sakamoto, M., Pinkel, D., Mascio, L., Sudar, D., Peters, D., Kuo, WL. L., Yamakawa, K., Nakamura, Y., Drabkin, H., Jericevic, Z., Smith, L., & Gray, J. W. (1995). Semiautomated DNA probe mapping using digital imaging microscopy: II. System performance. Cytometry, 19(1), 60-69. https://doi.org/10.1002/cyto.990190108