Publication: Flow cytometric immunophenotyping of lymphocyte subsets in samples that contain a high proportion of non‐lymphoid cells
Issued Date
1994-01-01
Resource Type
ISSN
10970320
01964763
01964763
Other identifier(s)
2-s2.0-0028625807
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Mahidol University
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SCOPUS
Bibliographic Citation
Cytometry. Vol.18, No.4 (1994), 199-208
Suggested Citation
Kovit Pattanapanyasat, Dennis E. Kyle, Pongsri Tongtawe, Kosol Yongvanitchit, Suthat Fucharoen Flow cytometric immunophenotyping of lymphocyte subsets in samples that contain a high proportion of non‐lymphoid cells. Cytometry. Vol.18, No.4 (1994), 199-208. doi:10.1002/cyto.990180403 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/9523
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Title
Flow cytometric immunophenotyping of lymphocyte subsets in samples that contain a high proportion of non‐lymphoid cells
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Abstract
Flow cytometric (FCM) immunophenotyping of peripheral blood from thalassemia patients presents technical difficulties because of the high proportion of immature red cells. The combination of forward scatter (FSC) and side scatter (SSC) with fluorescence associated with human leukocyte antigen/monocyte antigen (CD45/CD14) was unable to identify the lymphocyte population in thalassemia patients; therefore, it was necessary to exclude immature red cells to analyze lymphocyte subsets in these patients. A simultaneous three‐color FCM method was developed, with the basis that transferrin receptor (CD71) or glycophorin A (glyco A) is present on all immature red cells, but is not expressed on CD45 positive leukocytes. In this study, the lymphocyte population was identified by gating out unwanted cell populations using the FSC/CD71‐flurescein isothiocyanate (FITC), FSC/glyco A‐FITC, or FSC/CD45‐peridinin chlorophyl protein (PerCP) profiles. The CD71‐FITC negative cells, glyco A‐FITC negative cells, or CD45‐PerCP positive cells were identified, then analyzed on the basis of FSC/SSC to allow any remaining non‐lymphocyte cells in FSC/SSC gate to be excluded. The cells in FSC/SSC gate were then analyzed using other irrelevant two‐color antibodies. Of the three gating strategies, CD45‐PerCP and glyco A‐FITC methods are better than the CD71‐FITC gating method. Both methods markedly increase the purity of lymphocytes in the analysis gate. Either method is easy, straightforward, requires a six‐tube set of reagent tubes, and provides a reliable method for immunophenotyping lymphocyte subsets in preparations containing a large percentage of nonlymphoid cells. © 1994 Wiley‐Liss, Inc. Copyright © 1994 Wiley‐Liss, Inc.