Semiautomated DNA probe mapping using digital imaging microscopy

II. System performance

M. Sakamoto, D. Pinkel, L. Mascio, D. Sudar, D. Peters, W. L. Kuo, K. Yamakawa, Y. Nakamura, H. Drabkin, Z. Jericevic, L. Smith, Joe Gray

Research output: Contribution to journalArticle

19 Citations (Scopus)

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 PLpter 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.

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

Fingerprint

Cosmids
DNA Probes
Bacteriophages
Microscopy
Chromosomes
Chromosomes, Human, Pair 3
Telomere
Metaphase
Fluorescence In Situ Hybridization

Keywords

  • Digital microscopy
  • FISH
  • FLpter
  • Image analysis
  • Mapping

ASJC Scopus subject areas

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

Cite this

Semiautomated DNA probe mapping using digital imaging microscopy : II. System performance. / Sakamoto, M.; Pinkel, D.; Mascio, L.; Sudar, D.; Peters, D.; Kuo, W. L.; Yamakawa, K.; Nakamura, Y.; Drabkin, H.; Jericevic, Z.; Smith, L.; Gray, Joe.

In: Cytometry, Vol. 19, No. 1, 1995, p. 60-69.

Research output: Contribution to journalArticle

Sakamoto, M, Pinkel, D, Mascio, L, Sudar, D, Peters, D, Kuo, WL, Yamakawa, K, Nakamura, Y, Drabkin, H, Jericevic, Z, Smith, L & Gray, J 1995, 'Semiautomated DNA probe mapping using digital imaging microscopy: II. System performance', Cytometry, vol. 19, no. 1, pp. 60-69. https://doi.org/10.1002/cyto.990190108
Sakamoto, M. ; Pinkel, D. ; Mascio, L. ; Sudar, D. ; Peters, D. ; Kuo, W. L. ; Yamakawa, K. ; Nakamura, Y. ; Drabkin, H. ; Jericevic, Z. ; Smith, L. ; Gray, Joe. / Semiautomated DNA probe mapping using digital imaging microscopy : II. System performance. In: Cytometry. 1995 ; Vol. 19, No. 1. pp. 60-69.
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