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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.
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Updated: Dec 26, 2025

High-Dimensionality Flow Cytometry for Immune Function Analysis of Dissected Implant Tissues

High-Dimensionality Flow Cytometry for Immune Function Analysis of Dissected Implant Tissues

Published on: September 15, 2021

Panel Design and Optimization for High-Dimensional Immunophenotyping Assays Using Spectral Flow Cytometry.

Laura Ferrer-Font¹, Christophe Pellefigues¹, Johannes U Mayer¹

¹Malaghan Institute of Medical Research, Wellington, New Zealand.

Current Protocols in Cytometry

|March 10, 2020

Summary

This summary is machine-generated.

Spectral flow cytometry advances immune cell analysis by distinguishing full fluorophore emission spectra, enabling more flexible, high-dimensional panel design. This guide offers a step-by-step protocol for building and optimizing these complex panels.

Keywords:

fluorescent antibodies high-dimensional panel design immunophenotyping optimization spectral flow cytometry

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Area of Science:

Immunology
Biotechnology
Analytical Chemistry

Background:

Advances in fluorescence flow cytometry and immunology have driven the development of complex, high-parameter panels.
Designing and analyzing high-dimensional flow cytometry data presents significant challenges.

Purpose of the Study:

To introduce spectral flow cytometry as an advancement over conventional methods.
To provide a comprehensive protocol for designing and optimizing high-dimensional spectral flow cytometry panels.

Main Methods:

Spectral flow cytometry distinguishes the full emission spectrum of each fluorophore, unlike conventional methods that identify only peak emission.
This allows for greater flexibility in panel design and fluorophore selection, even with overlapping dyes.

Main Results:

Spectral flow cytometry accommodates fluorophores with similar emission maxima but distinct off-peak signatures.
The study provides a step-by-step protocol for building, designing, and optimizing high-dimensional spectral flow cytometry panels.

Conclusions:

Spectral flow cytometry offers enhanced capabilities for high-dimensional analysis of complex biological systems like the immune system.
The provided protocol guides users in overcoming challenges associated with complex panel design and fluorophore selection.