Open AccessEnzymatic amplification staining for flow cytometric analysis of cell surface molecules
Author(s) -
Kaplan David,
Smith Dawn
Publication year - 2000
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(20000501)40:1<81::aid-cyto11>3.0.co;2-k
Subject(s) - flow cytometry , microbiology and biotechnology , biology , staining , cd3 , cell , mhc class i , cluster of differentiation , phosphatidylserine , major histocompatibility complex , chemistry , biochemistry , cd8 , antigen , immunology , genetics , membrane , phospholipid
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. Cytometry 40:81–85, 2000 © 2000 Wiley‐Liss, Inc.