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Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry
Published on: March 5, 2019
Multicolor Digital Flow Cytometry in Human Translational Immunology
Samit R Joshi1,2, Subhasis Mohanty3, Albert C Shaw4
1Section of Infectious Diseases, Department of Internal Medicine, Yale School of Medicine, 300 Cedar St., 208056, New Haven, CT, 06520, USA. samit.joshi@yale.edu.
Insights
Multicolor flow cytometry enables simultaneous analysis of diverse cell types, crucial for immunology studies with limited samples. Standardizing instrument performance ensures accurate data for human cohort research.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Multicolor flow cytometry is vital for analyzing diverse cell lineages and biological pathways in human immunology.
- Limited sample amounts in studies, especially with older individuals, necessitate efficient analytical methods.
- Reproducible instrument standardization is critical for accurate comparisons in heterogeneous human subject groups.
Purpose of the Study:
- To describe procedures for multicolor digital flow cytometry.
- To share experience in flow cytometry panel design.
- To present an approach for standardizing instrument performance.
Main Methods:
- Utilizing multicolor digital flow cytometry techniques.
- Implementing standardized fluorochrome panel design.
- Applying procedures for instrument performance standardization using BD Biosciences hardware and software.
Main Results:
- Generation of accurate and precise data is achievable.
- Techniques are applicable in various research settings.
- Standardization ensures reliable results for human cohort studies.
Conclusions:
- The described procedures facilitate accurate and precise multicolor flow cytometry.
- Standardization of instrument performance is key for reliable human immunology research.
- These methods support robust data generation in diverse research environments.
Abstract:
By facilitating the simultaneous analysis of parameters from diverse cell lineages and biological pathways, multicolor flow cytometry is integral to many studies in human immunology-particularly those in older individuals-where sample amounts may be limiting. Studies in human cohorts require particular attention to fluorochrome panel design and procedures to standardize instrument performance; reproducible instrument conditions (over time and between centers) are crucial to accurate comparisons and conclusions in the analysis of heterogeneous groups of human subjects. Here, we describe procedures for multicolor digital flow cytometry, our experience in flow cytometry panel design and our approach in standardizing instrument performance using BD Biosciences hardware and software (BD Biosciences, San Jose, CA). These techniques allow for the generation of accurate and precise data in a variety of settings.
