title: Collection, Storage, and Preparation of Human Blood Cells.
abstract: Human peripheral blood is often studied by flow cytometry in both the research and clinical laboratories. The methods for collection, storage, and preparation of peripheral blood will vary depending on the cell lineage to be examined as well as the type of assay to be performed. This unit presents protocols for collection of blood, separation of leukocytes from whole blood by lysis of erythrocytes, isolating mononuclear cells by density gradient separation, and assorted non-flow sorting methods, such as magnetic bead separations, for enriching specific cell populations, including monocytes, T lymphocytes, B lymphocytes, neutrophils, and platelets, prior to flow cytometric analysis. A protocol is also offered for cryopreservation of cells, since clinical research often involves retrospective flow cytometric analysis of samples stored over a period of months or years.
authors.affiliation: National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Dagur
authors.affiliation: National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland.
authors.forename: J Philip
authors.initials: JP
authors.lastname: McCoy
mesh_terms: D000643:Ammonium Chloride / Q000494:pharmacology; D000906:Antibodies / Q000378:metabolism; D000925:Anticoagulants / Q000494:pharmacology; D001773:Blood Cells / Q000166:cytology* / Q000187:drug effects; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects; D001800:Blood Specimen Collection / Q000379:methods*; D002448:Cell Adhesion / Q000187:drug effects; D002458:Cell Fractionation; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D003165:Complement System Proteins / Q000378:metabolism; D015925:Cryopreservation; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects; D006801:Humans; D007202:Indicators and Reagents; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D060328:Magnetic Phenomena; D008863:Microspheres; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D010969:Plastics / Q000494:pharmacology; D011309:Preservation, Biological / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D000906:Antibodies; D000925:Anticoagulants; D007202:Indicators and Reagents; D010969:Plastics; D000643:Ammonium Chloride; D003165:Complement System Proteins
keywords: cell population isolation; cell population purification; flow cytometry; human blood; leukocyte preparation; lymphocyte enrichment; mononuclear blood cells