Enzymatic amplification staining for flow cytometric analysis of cell surface molecules
D Kaplan, D Smith - Cytometry: The Journal of the International …, 2000 - Wiley Online Library
D Kaplan, D Smith
Cytometry: The Journal of the International Society for Analytical …, 2000•Wiley Online LibraryBackground 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 …
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 …
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.
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