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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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Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Multiplexed cytometry for single cell chemical biology.

Henry A M Schares1, Madeline J Hayes2, Joseph A Balsamo3

  • 1Department of Chemistry, Vanderbilt University, Nashville, TN, United States; Vanderbilt Institute of Chemical Biology, Nashville, TN, United States; Vanderbilt Chemical and Physical Biology Program, Vanderbilt University, Nashville, TN, United States.

Methods in Cell Biology
|April 3, 2025
PubMed
Summary
This summary is machine-generated.

This protocol bridges chemistry and biology for chemical biology discovery using flow cytometry. It enables novice researchers to screen small molecules and cell types for functional readouts.

Keywords:
Chemical biologyCompound screeningFlow cytometryFluorescent cell barcodingMetabolomicsNatural product discoverySingle cellTraining protocol

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Area of Science:

  • Chemical Biology
  • Cell Biology
  • Translational Research

Background:

  • Flow cytometry offers deep cellular quantification and high throughput, ideal for translational research.
  • Underutilization in chemical biology stems from expertise gaps between chemists and cell biologists.
  • Bridging this gap requires accessible protocols for novice researchers.

Purpose of the Study:

  • To provide a detailed protocol bridging chemistry and biology for chemical biology discovery.
  • To train novice researchers in functional cell-based cytometry assays.
  • To encourage the use of flow cytometry in discovery and translational research.

Main Methods:

  • Presents a protocol adaptable for small molecule inputs (microbial extracts, pure compounds) and diverse cell types (primary human, mouse, cancer cell lines).
  • Details cytometry readouts for cell function, including DNA damage response, growth, apoptosis, cell cycle, and viability.
  • Incorporates 96-well plate format, fluorescent cell barcoding, and the debarcodeR algorithm for streamlined analysis.

Main Results:

  • The protocol enables selection of inputs, target cells, and cytometry readout modules.
  • Demonstrates adaptability for various functional readouts like cell identity, differentiation, and injury mechanisms.
  • Facilitates deep quantification of cellular features for millions of cells rapidly.

Conclusions:

  • This protocol empowers chemical biologists to integrate functional cell-based cytometry into discovery pipelines.
  • It democratizes the use of advanced flow cytometry techniques for a broader research community.
  • Aims to foster innovation in chemical biology through accessible, high-throughput screening methods.