An automated method to prepare cell suspensions from human biopsy samples for immunophenotyping by flow cytometry.
A comparison was made between a manual and automated method for preparing single cell suspensions from various types of human biopsy samples. The automated method uses the shearing action of fluid movement generated by a reciprocating paddle system. When compared on 11 samples, the automated instrument always isolated cells in less time, usually requiring only 15-30 seconds in contrast to 10-15 minutes for the manual method. The time comparisons involved "set up" time of the required minor equipment as well as the time to make a complete single cell suspension from the portion of the biopsy sample sent to the Flow Cytometry Lab so extra cells could be used for other purposes and cryogenically stored for future reference. Cells isolated by the automated method from various B-lymphocytic and T-lymphocytic malignancies were still viable and could be successfully immunophenotyped by flow cytometry. The immunophenotyping results were compared for both cell isolation methods on seven of these samples, and the results were comparable. The automated technique has now been used to satisfactorily immunophenotype more than 50 biopsy samples. The automated method should represent a significant aid for clinical flow cytometry laboratories performing immunophenotyping tests by efficiently preparing single cell suspensions in a few seconds instead of minutes. Furthermore, the automated method uses a closed sterile bag system that helps minimize the exposure of personnel to potential infectious material present in biopsy samples and prevents external contamination of cell suspensions. The automated technique proven successful for immunophenotyping may also be helpful for related procedures involving hematopoietic malignancies such as DNA content analysis and cell functional assays by flow cytometry, as well as other assays such as tissue typing, gene probe, and in vitro chemosensitivity assays.
Baystate Medical Center, Springfield, Massachusetts 01199.
M J
MJ
Warzynski
A E
AE
Podgurski
D M
DM
Boldt
R N
RN
Otto
D015703:Antigens, CD / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis*; D001322:Autoanalysis; D001706:Biopsy; D003584:Cytological Techniques; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D007118:Immunoassay; D007938:Leukemia / Q000473:pathology; D008223:Lymphoma / Q000473:pathology; D010641:Phenotype
D015703:Antigens, CD; D000954:Antigens, Surface; D005456:Fluorescent Dyes