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Laser excitation power and the flow cytometric resolution of complex karyotypes
Author(s) -
Ng Bee L.,
Carter Nigel P.
Publication year - 2010
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20904
Subject(s) - karyotype , chromosome , laser , biology , resolution (logic) , cell culture , microbiology and biotechnology , genetics , optics , physics , computer science , artificial intelligence , gene
The analytical resolution of individual chromosome peaks in the flow karyotype of cell lines is dependent on sample preparation and the detection sensitivity of the flow cytometer. We have investigated the effect of laser power on the resolution of chromosome peaks in cell lines with complex karyotypes. Chromosomes were prepared from a human gastric cancer cell line and a cell line from a patient with an abnormal phenotype using a modified polyamine isolation buffer. The stained chromosome suspensions were analyzed on a MoFlo sorter (Beckman Coulter) equipped with two water‐cooled lasers (Coherent). A bivariate flow karyotype was obtained from each of the cell lines at various laser power settings and compared to a karyotype generated using laser power settings of 300 mW. The best separation of chromosome peaks was obtained with laser powers of 300 mW. This study demonstrates the requirement for high‐laser powers for the accurate detection and purification of chromosomes, particularly from complex karyotypes, using a conventional flow cytometer. © 2010 International Society for Advancement of Cytometry