title: Karyotyping human and mouse cells using probes from single-sorted chromosomes and open source software.
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.
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Tamara A
authors.initials: TA
authors.lastname: Potapova
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Jay R
authors.initials: JR
authors.lastname: Unruh
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Box
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: William D
authors.initials: WD
authors.lastname: Bradford
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Seidel
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Slaughter
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Shamilene
authors.initials: S
authors.lastname: Sivagnanam
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.
authors.forename: Yuping
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO.|Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS.|Department of Cell Biology, Johns Hopkins University School of Medicine, 855, N. Wolfe Street, 21205, Baltimore, MD.
authors.forename: Rong
authors.initials: R
authors.lastname: Li
mesh_terms: D000818:Animals; D002460:Cell Line; D002877:Chromosomes, Human / Q000737:chemistry; D033481:Chromosomes, Mammalian / Q000737:chemistry*; D006801:Humans; D007621:Karyotyping / Q000379:methods*; D051379:Mice; D012984:Software
keywords: Image J; aneuploidy; chromosomes; cytogenetics; karyotyping; spectral unmixing