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

Journal Article · Sun Jan 01 04:00:00 EST 1995 · Cytometry

DOI:https://doi.org/10.1002/cyto.990190108· OSTI ID:254401

Sakamoto, M; Pinkel, D; Sudar, D; Peters, D ^[1]

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1. Univ. of California, San Francisco, CA (United States); and others

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. 27 refs., 4 figs., 3 tabs.

• DOE Contract Number: W-7405-ENG-48; AC03-76SF00098
• OSTI ID: 254401
• Journal Information: Cytometry, Journal Name: Cytometry Journal Issue: 1 Vol. 19; ISSN CYTODQ; ISSN 0196-4763
• Country of Publication: United States
• Language: English

Related Subjects

44 INSTRUMENTATION
INCLUDING NUCLEAR AND PARTICLE DETECTORS
55 BIOLOGY AND MEDICINE
BASIC STUDIES
99 GENERAL AND MISCELLANEOUS
AUTOMATION
COSMIDS
DIGITAL SYSTEMS
DNA
DNA SEQUENCING
DNA-CLONING
EFFICIENCY
FLUORESCENCE
GENETIC MAPPING
HUMAN CHROMOSOME 3
HUMAN CHROMOSOMES
IMAGE PROCESSING
MICROSCOPY
PERFORMANCE
PROBES
RESOLUTION
TELOMERES