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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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Related Experiment Video

Updated: Jun 9, 2025

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

Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi-pass

Marissa D Fahlberg¹, Sarah Forward¹, Emane Rose Assita¹

¹LASE Innovation Inc., Waltham, Massachusetts, USA.

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

|October 28, 2024

Summary

This study introduces a new multi-pass flow cytometry method. It allows accurate measurement of fragile cell markers before chemical fixation, improving data quality for cell biology research.

Keywords:

fixation and permeabilization flow cytometry fluorescent protein intracellular assays laser particles optical barcoding phospho‐flow single cell

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Last Updated: Jun 9, 2025

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

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Published on: April 14, 2015

Area of Science:

Cell Biology
Immunology
Biotechnology

Background:

Cell fixation and permeabilization are crucial for intracellular biomarker analysis in flow cytometry.
These chemical treatments can damage sensitive targets like fluorescent proteins and labels, compromising data accuracy.
Conventional flow cytometry offers a single measurement opportunity, limiting comprehensive cellular analysis.

Purpose of the Study:

To develop a novel multi-pass flow cytometry technique for preserving fragile cellular targets.
To enable sequential analysis of the same cells, maintaining single-cell resolution.
To overcome the limitations of conventional flow cytometry in handling chemically sensitive markers.

Main Methods:

Individual cell barcoding using laser particles for sequential analysis of single cells.
Measurement of chemically-fragile markers and fluorochromes before fixation and permeabilization.
Subsequent measurement of intracellular markers after chemical processing.

Main Results:

Accurate measurement of intracellular fluorescent proteins (FPs) and methanol-sensitive antigens/fluorophores.
Successful analysis of both surface and intracellular markers without compromising data integrity.
Demonstration of enhanced assay flexibility and comprehensive cellular analysis.

Conclusions:

The multi-pass flow cytometry approach preserves fragile cellular targets and fluorochromes.
This technique significantly improves accuracy and flexibility in flow cytometry workflows.
The innovation offers broad applications in immuno-oncology, stem cell research, and cell biology.