Mammalian Chromosome Analysis and Sorting by Flow Cytometry

Mukhopadhyay, Risani; Varshitha, D. V.; Telford, William G.; Sanders, Claire Kathleen; Chakraborty, Uttara

doi:10.1002/cpz1.785

Mammalian Chromosome Analysis and Sorting by Flow Cytometry

Journal Article · Thu May 18 00:00:00 EDT 2023 · Current Protocols

DOI:https://doi.org/10.1002/cpz1.785· OSTI ID:2432306

Mukhopadhyay, Risani ^[1]; Varshitha, D. V. ^[2]; Telford, William G. ^[3]; ^[4]; Chakraborty, Uttara ^[1]

Manipal Academy of Higher Education (India)
Bengaluru (India)
National Cancer Institute, Bethesda, MD (United States)
Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)

The analysis of chromosomes by flow cytometry is termed flow cytogenetics, and it involves the analysis and sorting of single mitotic chromosomes in suspension. The study of flow karyograms provides insight into chromosome number and structure to provide information on chromosomal DNA content and can enable the detection of deletions, translocations, or any forms of aneuploidy. Beyond its clinical applications, flow cytogenetics greatly contributed to the Human Genome Project through the ability to sort pure populations of chromosomes for gene mapping, cloning, and the construction of DNA libraries. Maximizing the potential of these important applications of flow cytogenetics relies on precise instrument setup and optimal sample processing, both of which impact the accuracy and quality of the data that are generated. This article is a compilation of the existing protocols that describe the stepwise methodology of accumulating, isolating, and staining metaphase chromosomes to prepare single-chromosome suspensions for flow cytometric analysis and sorting. Although the chromosome preparation protocols have remained largely unchanged, cytometer technology has advanced dramatically since these protocols were originally developed. Advances in cytometry technologies offer new and exciting approaches for understanding and monitoring chromosomal aberrations, but the hallmark of these protocols remains their simplicity in methodologies and reagent requirements and the accuracy of data resolvable to every chromosome of the cell.

View Accepted Manuscript (DOE)

Research Organization:: Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)

Sponsoring Organization:: USDOE Laboratory Directed Research and Development (LDRD) Program; USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation. Bioenergy Technologies Office (BETO)

Grant/Contract Number:: 89233218CNA000001

OSTI ID:: 2432306

Report Number(s):: LA-UR--23-20462

Journal Information:: Current Protocols, Journal Name: Current Protocols Journal Issue: 5 Vol. 3; ISSN 2691-1299

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
flow cytogenetics
isolation protocols
karyotype
mammalian chromosomes
sorting

Mammalian Chromosome Analysis and Sorting by Flow Cytometry

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References (26)

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