UNIT 5.6 Measurement of Intercellular Conjugates by Flow Cytometry
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Segal, D. M. 2001. Measurement of Intercellular Conjugates by Flow Cytometry. Current Protocols in Immunology. 64:5.6:5.6.1–5.6.8.
- Published Online: 1 MAY 2001
- Published Print: JUN 1995
Analysis of fluorescently labeled cellular aggregates can be accomplished quickly and accurately with the technique of flow cytometry. With a dual-laser flow cytometer, the excitation and emission spectra of fluorophores used to label cells are widely separated, and there is no spillover of emission from one fluorophore into the detector measuring emission from the other. Consequently, conjugates can be measured by a dual-laser instrument when the extent of labeling two cell types with fluorophores are very different. The more commonly available single-laser cytometers can also be used to measure multicellular conjugates, but due to overlaps in emission spectra, the extent of labeling cells with fluorophores must be controlled much more carefully when the single-laser machines are used. This unit describes the labeling of cells and analysis of conjugates with either dual-laser or single laser flow cytometers.