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Flow Cytometry and Its Utility

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Chromosome Structure and Aberrations

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Abstract

Flow cytometry is a laser-based biophysical technique to study optical properties of microscopic particles in fluid suspension. The technology is not only routinely used in medicine (transplantation, hematology, cancer, prenatal diagnosis, genetics, and sperm sorting for sex preselection) but also has many other applications in basic research, crop improvement, clinical practice, and clinical trials. The principles of flow cytometric chromosome analysis and sorting known as flow cytogenetics and research in this field have increased over the past four decades due to high sensitivity and precision of this technique. The use of nucleic acid-specific fluorochromes has semiautomated quantitative chromosome analysis, thus reducing subjectivity of preexisting slide-based methods. Flow cytometric classification of chromosomes (flow karyotyping) enables detection of chromosomal aberrations, while flow sorting permits isolation of single chromosome types in large quantities to be used for gene mapping, preparation of chromosome-specific gene libraries, and ultimately sequencing leading to chromosome genomics. Flow fluorescent in situ hybridization (FISH), i.e., combined fluorescent in situ hybridization in suspension (FISHIS) and flow cytometry using microsatellite DNA or gene-specific probes, is another approach to increase our knowledge on structure, function, and evolution of chromosomes. These techniques provide increased value for diagnosis and management of clinical diseases, especially cancer which is generally characterized by high chromosomal instability. Flow cytogenetics also has important applications in plant biology, especially in the study of genome evolution and crop improvement. This review outlines the utility of flow cytometry in chromosomal analysis, its applications, limitations, and future directions.

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  1. Center of Advanced Study, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, 700 019, West Bengal, India

    Mihir Halder, Sayantani Nath & Sumita Jha

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  2. Sayantani Nath
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Editors and Affiliations

  1. Department of Education, Government of Jammu and Kashmir, Srinagar, India

    Tariq Ahmad Bhat

  2. Department of Botany, University of Kashmir, Srinagar, Jammu and Kashmir, India

    Aijaz Ahmad Wani

Glossary

Allopolyploid

Polyploidy with more than one basic set of chromosomes

Apoptosis

Biological event of program cell death observed in multicellular organism

Autofluorescence

Intrinsic property of some biomolecules that emits fluorescence after excitation with light

B chromosome

Extra-small chromosomes over standard set of chromosomes

Chromosomal aberrations

Structural or numerical changes of chromosome

Chromosome painting

Chromosomal in situ hybridization with multiple sequence-specific fluorescent probes

Cytogenetics

Branch of genetics that deals with the study of structural and functional aspect of the cell, predominantly the chromosomes

Cytophotometry

The analysis of cellular content with the help of an instrument called cytophotometer

Deletion

Loss of a segment of a chromosome

DNA library

Collection of DNA fragments of an organism that was cloned into vectors

Flow cytometry

Laser-based biophysical technique to study optical properties of cells in liquid stream

Fluorescence in situ hybridization (FISH)

Modern technique used for detecting and locating a specific DNA sequence on a chromosome using probe

Fluorochrome

Chemical molecules that absorbed a particular wavelength of light, get excited and reemit light of higher wavelength in the form of fluorescent

Genetic marker

Unique DNA sequence used in molecular mapping

Genome

Complete set of DNA in an organism

PCR

Polymerase chain reaction

Probes

Small nucleic acid sequence (DNA or RNA) or monoclonal antibodies or fluorochrome-conjugated antibodies used for detection of specific molecule

Sex chromosome

Chromosomes responsible for the determination of sex of an organism

SSC

The light scattered 90° to the incident laser beam as the cell flow through the flow cytometer

Translocation

A chromosomal aberration that occurred due to the rearrangement of chromosomal parts in between two non-homologous chromosomes

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Halder, M., Nath, S., Jha, S. (2017). Flow Cytometry and Its Utility. In: Bhat, T., Wani, A. (eds) Chromosome Structure and Aberrations. Springer, New Delhi. https://doi.org/10.1007/978-81-322-3673-3_5

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