Pattanapanyasat, KPengruangrojanachai, VThepthai, CSuwanagool, SWasi, C2009-05-272009-05-271994-12-01Pattanapanyasat K, Pengruangrojanachai V, Thepthai C, Suwanagool S, Wasi C. Flow cytometric three-color determination of CD4 T-lymphocytes on blood specimens from AIDS patients who have a large number of contaminating non-lymphocytes. Asian Pacific Journal of Allergy and Immunology. 1994 Dec; 12(2): 105-9http://imsear.searo.who.int/handle/123456789/36566Published by the Allergy and Immunology Society of Thailand.A three-color flow cytometric determination of CD4 T-lymphocytes on whole blood specimens from AIDS patients which contain a high proportion of non-lymphocyte elements is described. Peripheral blood cells were stained by a three-color method using monoclonal antibodies conjugated respectively with fluorescein isothiocyanate (FITC)-CD3, phycoerythrin (PE)-CD4 and peridinin chlorophyll protein (PerCP)-CD45. CD45 stains all leukocytes with the highest fluorescence expression of CD45 antigen in lymphocytes. By combining light scatter with CD45 in the fluorescence 3 (FL3) channel, a light scattering window can be drawn to include almost all bright CD45 lymphocytes. This live gate of lymphocytes was then acquired and analysed simultaneously using other irrelevant two-color (FITC/PE) antibodies of CD3 and CD4 in the FITC and PE channels, respectively. This method is easy and straightforward, and gives successful analysis of CD4 T-lymphocytes in AIDS blood specimens contaminated with an unusually large number of non-lymphocytic cells.engAcquired Immunodeficiency Syndrome --bloodAntibodies, MonoclonalAntigens, CD3 --analysisAntigens, CD4 --analysisAntigens, CD45 --analysisCD4 Lymphocyte Count --methodsCD4-Positive T-Lymphocytes --immunologyFlow Cytometry --methodsFluorescent DyesHIV Seropositivity --bloodHumansImmunophenotypingMaleJournal Article