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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.
Immunology Today
|October 8, 2014
Summary
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.
