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

Area of Science:

• Genetics
• Molecular Biology
• Bioimaging

Background:

• Multispectral karyotyping is crucial for analyzing chromosomes but often requires expensive specialized equipment.
• Existing methods present a barrier to entry for many research laboratories.

Purpose of the Study:

• To develop a cost-effective multicolor karyotyping approach.
• To provide open-source software for spectral unmixing and karyotyping.

Main Methods:

• Generated multicolor chromosome paints from single-sorted human and mouse chromosomes.
• Developed the Karyotype Identification via Spectral Separation (KISS) analysis package, an open-source ImageJ toolset.
• Utilized spectral laser scanning confocal microscopy for imaging and analysis.

Main Results:

• Demonstrated the feasibility of using conventional spectral confocal microscopes for karyotyping.
• The KISS package successfully performs spectral unmixing and karyotype identification.
• Enabled multicolor karyotyping without specialized multispectral systems.

Conclusions:

• Multicolor karyotyping can be performed cost-effectively using accessible spectral microscopes and open-source software.
• The KISS package democratizes advanced chromosomal analysis for academic research.
• This approach lowers the financial barrier for multicolor karyotyping.