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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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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: Jul 9, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

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High-dimensional multi-pass flow cytometry via spectrally encoded cellular barcoding.

Sheldon J J Kwok1, Sarah Forward2, Marissa D Fahlberg2

  • 1LASE Innovation Inc., Woburn, MA, USA. skwok@laseinno.com.

Nature Biomedical Engineering
|November 30, 2023
PubMed
Summary
This summary is machine-generated.

Multi-pass high-dimensional flow cytometry uses cellular barcoding to repeatedly measure the same cells over time. This technique enhances single-cell analysis capabilities for immunology and drug discovery.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Flow cytometry advancements are crucial for immunology, immuno-oncology, drug discovery, and vaccine development.
  • Current limitations include spectral overlap in fluorophore markers and single-timepoint measurements.
  • Measuring more protein markers per cell over time is a key challenge.

Purpose of the Study:

  • To introduce multi-pass high-dimensional flow cytometry.
  • To enable repeated, high-parameter measurements of individual cells over time.
  • To overcome limitations of traditional flow cytometry for complex biological studies.

Main Methods:

  • Development of a novel cellular barcoding technique using microparticles emitting near-infrared laser light.
  • Application of multi-pass analysis, where individual cells are measured multiple times.
  • Utilizing live human peripheral blood mononuclear cells for method validation.

Main Results:

  • Demonstrated time-resolved characterization of cells before and after stimulation.
  • Enabled analysis with a 10-marker panel with minimal spectral compensation.
  • Achieved deep immunophenotyping using a 32-marker panel across 3 cycles (10-13 markers/cycle).

Conclusions:

  • Cellular barcoding in flow cytometry significantly extends the technique's utility.
  • The method facilitates high-dimensional, multi-pass single-cell analyses.
  • This approach simplifies marker-panel design and reduces spectral spillover.