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Updated: Apr 22, 2026
Fluorescence-activated Cell Sorting for Purification of Plasmacytoid Dendritic Cells from the Mouse Bone Marrow
Published on: November 4, 2016
Dual immunofluorescence - new frontiers in cell analysis and sorting
D R Parks1, R R Hardy, L A Herzenberg
1Genetics Department, Stanford University School of Medicine, Stanford, CA 94305, USA.
Insights
Fluorescence-activated cell sorting (FACS) and flow cytometry transformed immunology. However, single immunofluorescence measurements are insufficient for detailed lymphoid cell analysis, necessitating multiparameter approaches.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Fluorescence-activated cell sorting (FACS) and flow cytometry have revolutionized immune system research.
- Current limitations exist in analyzing complex lymphoid cell subpopulations with single immunofluorescence measurements.
Purpose of the Study:
- To review the necessity of multiparameter analysis in flow cytometry for detailed immune cell investigation.
- To highlight the limitations of single-labeling techniques in resolving functional lymphoid subsets.
Main Methods:
- Review of current literature on flow cytometry and immunofluorescence techniques.
- Discussion of the principles and applications of multiparameter analysis in cell sorting.
Main Results:
- Single immunofluorescence measurements provide inadequate resolution for many lymphoid cell subpopulation studies.
- Multiparameter analysis, using two or more fluorescent labels, is increasingly required for comprehensive functional subset definition.
Conclusions:
- Advanced flow cytometry techniques employing multiple fluorescent labels are essential for accurate characterization of immune cell heterogeneity.
- Future immunological investigations will increasingly rely on multiparameter flow cytometry for deeper insights into immune cell function.
Abstract:
Investigations of the nature and functions of the immune system and its cell populations have been revolutionized by the techniques of fluorescence-activated cell sorting (FA CS) and flow cytometry. Over the last few years, however, it has become increasingly clear that making only a single immunofluorescence measurement on each cell is not adequate for many investigations of lymphoid cell subpopulations. In this review the authors discuss why adequate resolution of functional subsets will increasingly require multiparameter definition including measurements on two or more fluorescent labels.
