A fluorometric approach to the quantitation of cell number with application to a cell adhesion assay

David Lewinsohn, Brian J. Nickoloff, Eugene C. Butcher

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

We describe a fluorometric approach to the determination of cell number. Cells are covalently labeled with fluorescein isothiocyanate (FITC) under physiologic conditions. Cells are lysed with detergent, and released fluorochrome is assessed quantitatively with a fluorescence spectrophotometer. Fluorescence varies linearly with regard to cell number over a wide range of concentrations, and allows detection of as few as 8 × 103 cells/ml. We describe the effect of different time and temperature incubations of FITC-labeled cells on fluorescence intensity, and we use the method to analyze the binding of peripheral blood mononuclear leukocytes to interferon-gamma-treated keratinocytes. Finally, we demonstrate that the results in an adhesion assay are comparable to those obtained with 51Cr-labeled cells. Quantitative fluorescence analysis of cell number offers a safe, inexpensive, rapid, and accurate method of determining cell number without the biological hazard and waste disposal problems associated with radioactive labeling.

Original languageEnglish (US)
Pages (from-to)93-100
Number of pages8
JournalJournal of Immunological Methods
Volume110
Issue number1
DOIs
StatePublished - May 25 1988
Externally publishedYes

Fingerprint

Cell Adhesion
Cell Count
Fluorescence
Fluorescein
Medical Waste Disposal
Mononuclear Leukocytes
Keratinocytes
Fluorescent Dyes
Interferon-alpha
Detergents
Interferon-gamma
Temperature
isothiocyanic acid

Keywords

  • Cell adhesion
  • Cell number
  • Fluorescein isothiocyanate
  • Fluorescence spectroscopy

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

A fluorometric approach to the quantitation of cell number with application to a cell adhesion assay. / Lewinsohn, David; Nickoloff, Brian J.; Butcher, Eugene C.

In: Journal of Immunological Methods, Vol. 110, No. 1, 25.05.1988, p. 93-100.

Research output: Contribution to journalArticle

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N2 - We describe a fluorometric approach to the determination of cell number. Cells are covalently labeled with fluorescein isothiocyanate (FITC) under physiologic conditions. Cells are lysed with detergent, and released fluorochrome is assessed quantitatively with a fluorescence spectrophotometer. Fluorescence varies linearly with regard to cell number over a wide range of concentrations, and allows detection of as few as 8 × 103 cells/ml. We describe the effect of different time and temperature incubations of FITC-labeled cells on fluorescence intensity, and we use the method to analyze the binding of peripheral blood mononuclear leukocytes to interferon-gamma-treated keratinocytes. Finally, we demonstrate that the results in an adhesion assay are comparable to those obtained with 51Cr-labeled cells. Quantitative fluorescence analysis of cell number offers a safe, inexpensive, rapid, and accurate method of determining cell number without the biological hazard and waste disposal problems associated with radioactive labeling.

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