Long-term preservation of whole blood samples for flow cytometry analysis in normal and HIV-infected individuals from Africa.
A whole blood method requiring less than 4 ml of heparinized blood was developed to assess the practicality of preparing whole blood samples that could be easily stored, transported and readily used to determine the lymphocyte phenotypes and proliferation responses of individuals from remote areas who are infected with the human immunodeficiency virus. Minor modifications in standard whole blood procedure for lymphocyte phenotyping have significantly increased the stability of light scatter and fluorescence intensity of the cells for subsequent flow cytometry (FC) analysis. These changes include removal of lysis solution prior to fixation, fixation of monoclonal antibody-stained cells in 1% paraformaldehyde for 30 minutes and storage of fixed samples in medium containing 1% bovine serum albumin. Lymphocyte subsets and their functional subsets could reliably be determined on samples stored for up to 4 weeks. Further, blood samples could be kept at room temperature for up to 96 hours or at ambient temperature during transportation from Africa before staining for FC without affecting their quantitation. While samples could be processed for FC analysis under field-laboratory conditions, proliferation assays could only be performed on samples that were transported within 48 hours of their collection. The whole blood method saves time and expense and decreases the volumes of blood required to perform phenotypic analysis and functional assays on specimens collected in remote areas.
Lal
R B
RB
Department of Preventive Medicine and Biometrics, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
Hira
S K
SK
Dhawan
R R
RR
Perine
P L
PL
D000328:Adult; D000349:Africa; D001793:Blood Preservation; D005260:Female; D005434:Flow Cytometry; D015658:HIV Infections / Q000097:blood* / Q000276:immunology; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D008213:Lymphocyte Activation; D008214:Lymphocytes; D008297:Male; D010641:Phenotype; D013194:Staining and Labeling; D013696:Temperature; D013997:Time Factors; D014186:Transportation
D006684:HLA-DR Antigens