Enzymatic amplification staining for flow cytometric analysis of cell surface molecules.
BACKGROUND
Flow cytometric analysis is a powerful technique for the single cell assessment of cell surface expression of selected molecules. The major deficiency of flow cytometry has been its relative insensitivity. Only molecules expressed in abundance have been readily observed.


METHODS
We have developed an enzymatic amplification procedure for the analysis of cell surface molecules by flow cytometry. Transformed and nontransformed cells expressing MHC class I, CD5, CD3, CD4, CD6, CD7, CD34, CD45, MHC class II, Fas ligand, and phosphatidylserine were assessed.


RESULTS
Our enzymatic amplification technology increased the fluorescence signal between 10 and 100-fold for all surface molecules tested.


CONCLUSIONS
Enzymatic amplification staining produces a significant enhancement in the resolving power of flow cytometric analysis of cell surface molecules. Using this technique, we have been able to detect the presence of molecules that could not be observed by the standard procedure.
Kaplan
D
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Department of Pathology, Case Western Reserve University, Cleveland, OH 44106-4943, USA. drk5@po.cwru.edu
Smith
D
D
D017304:Annexin A5 / Q000032:analysis / Q000276:immunology; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology; D018952:Antigens, CD34 / Q000032:analysis / Q000276:immunology; D019011:Antigens, CD7 / Q000032:analysis / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis*; D017209:Apoptosis; D017252:CD3 Complex / Q000032:analysis / Q000276:immunology; D015704:CD4 Antigens / Q000032:analysis / Q000276:immunology; D018956:CD5 Antigens / Q000032:analysis / Q000276:immunology; D002461:Cell Line, Transformed / Q000737:chemistry / Q000166:cytology; D004796:Clinical Enzyme Tests / Q000379:methods*; D053222:Fas Ligand Protein; D005434:Flow Cytometry / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000276:immunology; D006801:Humans; D019169:Jurkat Cells; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology; D010718:Phosphatidylserines / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity
D017304:Annexin A5; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018952:Antigens, CD34; D019011:Antigens, CD7; D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface; D017252:CD3 Complex; D015704:CD4 Antigens; D018956:CD5 Antigens; C062279:CD6 antigen; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D008562:Membrane Glycoproteins; D010718:Phosphatidylserines; D017493:Leukocyte Common Antigens