Flow microfluorometric studies of lectin binding to mammalian cells II. Estimation of the surface density of receptor sites by multiparameter analysis
✍ Scribed by John A. Steinkamp; Paul M. Kraemer
- Publisher
- John Wiley and Sons
- Year
- 1974
- Tongue
- English
- Weight
- 534 KB
- Volume
- 84
- Category
- Article
- ISSN
- 0021-9541
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✦ Synopsis
Abstract
A flow‐system multiparameter cell analyzer that simultaneously measures and processes fluorescence and cell volume signals from single cells was used to study the binding of fluorescein‐conjugated Concanavalin A (Con A–F) to the cell surface. Cells reacted with Con A–F were passed through a flow chamber where sensors measured both cell volume and fluorescence of each individual cell. Sensor signals were electronically processed by first converting the cell volume signals to two‐thirds power (proportional to surface area) and then forming the fluorescence‐to‐surface area ratio. These ratios, which were considered as estimates of the surface density of binding sites, were displayed as frequency distribution histograms using a multichannel pulse‐height analyzer for various cell populations differing in cell size. Comparisons between cell lines showed characteristic differences in binding site density. Cell cycle dependent changes were not found for CHO cells synchronized by mitotic selection. An important benefit of this analysis method was the ability to quantitate very weak cell surface fluorescence.