Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Flow Cytometry

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

Clonal Selection and Evolution after Treatment of Severe Aplastic Anemia.

NEJM evidence·2026

Same author

Imputing causality and clonal dynamics from single-cell transcriptomics in paroxysmal nocturnal hemoglobinuria.

Leukemia·2026

Same author

Selective Janus kinase inhibitors exhibit different effects in murine immune aplastic anemia.

HemaSphere·2026

Same author

T cell and monocyte activation in concert with hematopoietic stem cell interactions shapes the post-allogeneic transplant immune landscape in poor graft function.

Frontiers in immunology·2026

Same author

Reframing-renaming(?)-myelodysplastic syndromes/neoplasms and clonal hematopoiesis of indeterminate potential.

Leukemia·2026

Same author

STAT3 SH2 Domain Aspartic Acid 661 Mutations Activate Immune Gene Programs.

Journal of cellular and molecular medicine·2026

Search research articles

Related Experiment Video

Updated: Feb 27, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Optimization and standardization of fluorescent cell barcoding for multiplexed flow cytometric phenotyping.

Valentina Giudice¹, Xingmin Feng¹, Sachiko Kajigaya¹

¹Hematology Branch, National Heart, Lung, and Blood Institute (NHLBI), NIH, Bethesda, Maryland, 20892-1202.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|July 11, 2017

Summary

Optimized fluorescent cell barcoding (FCB) enhances multiplex flow cytometry for drug screening and T-cell analysis. This technique improves data robustness and reduces reagent use for high-throughput studies.

Keywords:

flow cytometry fluorescent cell barcoding phenotyping viability dye assay

More Related Videos

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Published on: October 4, 2018

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Published on: April 7, 2023

Related Experiment Videos

Last Updated: Feb 27, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Published on: October 4, 2018

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Published on: April 7, 2023

Area of Science:

Immunology
Biotechnology
Cell Biology

Background:

Fluorescent cell barcoding (FCB) is a multiplexing technique for high-throughput flow cytometry.
FCB enables collective staining and acquisition, reducing variability, antibody consumption, and sample volume.
Existing FCB techniques face technical challenges impacting routine use.

Purpose of the Study:

To optimize the FCB technique for routine application in biological research.
To enhance the robustness and efficiency of FCB for multiplex drug screening and lymphocyte characterization.
To adapt FCB for analyzing specific cell populations like T cells.

Main Methods:

Optimized FCB using DyLight 350, DyLight 800, Pacific Orange, and CBD500 dyes.
Tested working concentrations of FCB dyes from 0 to 500 microg/ml.
Optimized viability dye staining for improved data robustness.
Achieved five-color staining for Vβ usage analysis in CD4+ and CD8+ T cells with nine-sample barcoding.

Main Results:

Successfully barcoded six, nine, and 36 human peripheral blood specimens.
Established optimal dye concentrations and viability staining protocols.
Demonstrated feasibility of multiplex analysis of T-cell populations using FCB.
Improved FCB technique for increased robustness and wider applicability.

Conclusions:

The optimized FCB technique is suitable for routine use in high-throughput flow cytometry.
Improvements facilitate multiplex drug screening, signaling profiling, and cytokine detection.
Enhanced FCB is valuable for lymphocyte characterization and disease monitoring.