Assessment of endothelial immunophenotype - limitation of flow cytometric analysis

Kevin D. Forsyth, Vivienne Talbot

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Flow cytometry is generally utilized to quantify antigen expression by cells in suspension. To detect antigens on endothelium, which grows as a monolayer, either the creation of a suspension of endothelial cells for flow cytometry, or the use of alternative techniques (such as immunoperoxidase staining) is required. We demonstrate here that creating suspensions of endothelial cells for flow cytometry underestimates the expression of certain antigens. In addition, morphological information regarding certain antigens (serpins and fibronectin) is only discernible by immune microscopy, a subjective procedure. We would recommend caution in using flow cytometry for the estimation of endothelial antigens. Using computerized estimates of microscopic immunostaining (e.g., with video image analysis) it may be possible to overcome some of the subjective limitations of immune microscopy.

Original languageEnglish
Pages (from-to)93-99
Number of pages7
JournalJournal of Immunological Methods
Volume144
Issue number1
DOIs
Publication statusPublished - 1991
Externally publishedYes

Keywords

  • Antigen
  • Endothelium
  • Expression
  • Flow cytometry
  • Immunoperoxidase

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