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Biotechniques. 2015 Dec 1;59(6):335-6, 338, 340-2 passim. doi: 10.2144/000114362. eCollection 2015 Dec.

Karyotyping human and mouse cells using probes from single-sorted chromosomes and open source software.

Author information

1
Stowers Institute for Medical Research, Kansas City, MO.
2
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS.
3
Department of Cell Biology, Johns Hopkins University School of Medicine, 855, N. Wolfe Street, 21205, Baltimore, MD.

Abstract

Multispectral karyotyping analyzes all chromosomes in a single cell by labeling them with chromosome-specific probes conjugated to unique combinations of fluorophores. Currently available multispectral karyotyping systems require the purchase of specialized equipment and reagents. However, conventional laser scanning confocal microscopes that are capable of separating multiple overlapping emission spectra through spectral imaging and linear unmixing can be utilized for classifying chromosomes painted with multicolor probes. Here, we generated multicolor chromosome paints from single-sorted human and mouse chromosomes and developed the Karyotype Identification via Spectral Separation (KISS) analysis package, a set of freely available open source ImageJ tools for spectral unmixing and karyotyping. Chromosome spreads painted with our multispectral probe sets can be imaged on widely available spectral laser scanning confocal microscopes and analyzed using our ImageJ tools. Together, our probes and software enable academic labs with access to a laser-scanning spectral microscope to perform multicolor karyotyping in a cost-effective manner.

KEYWORDS:

Image J; aneuploidy; chromosomes; cytogenetics; karyotyping; spectral unmixing

PMID:
26651513
DOI:
10.2144/000114362
[Indexed for MEDLINE]
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