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Proceedings Paper

Flow analysis of human chromosome sets by means of mixing-stirring device
Author(s): Valeri V. Zenin; Nicolay D. Aksenov; Alla N. Shatrova; Nicolay V. Klopov; L. Scott Cram; Andrey I. Poletaev
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Paper Abstract

A new mixing and stirring device (MSD) was used to perform flow karyotype analysis of single human mitotic chromosomes analyzed so as to maintain the identity of chromosomes derived from the same cell. An improved method for cell preparation and intracellular staining of chromosomes was developed. The method includes enzyme treatment, incubation with saponin and separation of prestained cells from debris on a sucrose gradient. Mitotic cells are injected one by one in the MSD which is located inside the flow chamber where cells are ruptured, thereby releasing chromosomes. The set of chromosomes proceeds to flow in single file fashion to the point of analysis. The device works in a stepwise manner. The concentration of cells in the sample must be kept low to ensure that only one cell at a time enters the breaking chamber. Time-gated accumulation of data in listmode files makes it possible to separate chromosome sets comprising of single cells. The software that was developed classifies chromosome sets according to different criteria: total number of chromosomes, overall DNA content in the set, and the number of chromosomes of certain types. This approach combines the high performance of flow cytometry with the advantages of image analysis. Examples obtained with different human cell lines are presented.

Paper Details

Date Published: 2 May 1997
PDF: 8 pages
Proc. SPIE 2982, Optical Diagnostics of Biological Fluids and Advanced Techniques in Analytical Cytology, (2 May 1997); doi: 10.1117/12.273640
Show Author Affiliations
Valeri V. Zenin, Institute of Cytology (Russia)
Nicolay D. Aksenov, Institute of Cytology (Russia)
Alla N. Shatrova, Institute of Cytology (Russia)
Nicolay V. Klopov, St. Petersburg Institute of Nuclear Physics (Russia)
L. Scott Cram, Los Alamos National Lab. (United States)
Andrey I. Poletaev, Engelhardt Institute of Molecular Biology (Russia)

Published in SPIE Proceedings Vol. 2982:
Optical Diagnostics of Biological Fluids and Advanced Techniques in Analytical Cytology
Robert C. Leif; Alexander V. Priezzhev; Toshimitsu Asakura; Robert C. Leif, Editor(s)

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