Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
In...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Alpha-synuclein at the crossroads of host-virus interactions: immunological roles beyond the nervous system.

Journal of virology·2026
Same author

Distinct in vivo dynamics of donor-derived stem cell memory CAR T cells post-allogeneic HSCT relapse.

Cell·2026
Same author

Immunophenotypic skewing of B cells toward IgD⁻CD27⁻IgG⁺ subtype and metabolic attenuation in colorectal cancer.

Scientific reports·2026
Same author

Guidelines for T cell nomenclature.

Nature reviews. Immunology·2025
Same author

Single-Cell Atlas of Cardiac Endothelial Cell Heterogeneity in Pressure Overload.

Circulation research·2025
Same author

Immunosuppressive contribution of tumour-infiltrating B cells in human intrahepatic cholangiocarcinoma and their role in chemoimmunotherapy outcome.

Gut·2025

Related Experiment Video

Updated: Jun 27, 2026

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry
07:39

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Published on: January 7, 2019

Investigating T cells by polychromatic flow cytometry.

Enrico Lugli1, Leonarda Troiano, Andrea Cossarizza

  • 1Department of Biomedical Sciences, Section of General Pathology, Modena, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|December 3, 2008
PubMed
Summary

Polychromatic flow cytometry (PFC) enables detailed analysis of immune cells by measuring up to 19 parameters. This chapter discusses PFC protocols and data interpretation for complex immunological studies.

More Related Videos

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells
06:22

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Published on: January 9, 2019

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
09:04

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Published on: April 19, 2017

Related Experiment Videos

Last Updated: Jun 27, 2026

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry
07:39

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Published on: January 7, 2019

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells
06:22

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Published on: January 9, 2019

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
09:04

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

Published on: April 19, 2017

Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Flow cytometry has significantly advanced immunology by enabling single-cell analysis.
  • It facilitates the identification of immune cell subsets involved in immunoregulation and disease pathogenesis.
  • Recent technological advancements allow for multi-parameter analysis, up to 19 at the single-cell level.

Purpose of the Study:

  • To discuss protocols for performing polychromatic flow cytometry (PFC).
  • To provide guidelines for the interpretation and analysis of complex PFC data.
  • To highlight the utility of PFC in fine analysis of immune cell phenotypes.

Main Methods:

  • Utilizing advanced flow cytometry equipment and fluorochromes.
  • Implementing polychromatic flow cytometry (PFC) for simultaneous multi-parameter analysis (up to 19).
  • Developing protocols for PFC execution and data handling.

Main Results:

  • PFC allows for fine analysis of immune cell phenotypes, including T lymphocytes, B cell subsets, and intracellular phosphoproteome.
  • The approach generates large datasets, necessitating specialized interpretation strategies.
  • Established protocols and guidelines aid in managing and understanding complex PFC data.

Conclusions:

  • Polychromatic flow cytometry is a powerful tool for in-depth immunological research.
  • Effective data interpretation and analysis strategies are crucial for leveraging PFC's potential.
  • This chapter provides essential guidance for researchers utilizing PFC in immunology.