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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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Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media
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Characterization of Extracellular Vesicles by Flow Cytometry.

Virginia Camacho1, Vasilis Toxavidis1, John C Tigges2,3

  • 1Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 23, 2017
PubMed
Summary

This study presents a standardized nanoscale flow cytometry method for extracellular vesicle (EV) analysis. This approach ensures accurate EV detection, analysis, and sorting for reliable quality control and cross-platform comparison.

Keywords:
Extracellular vesiclesFlow cytometry methodsNanoparticle sortingNanoscaleSmall particle detectionSubmicron

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Extracellular vesicles (EVs) are crucial biomarkers, but their analysis requires standardized methods.
  • Nanoscale flow cytometry offers potential for high-throughput EV characterization.
  • Current methods lack standardization, hindering cross-platform comparability and reliable quality control.

Purpose of the Study:

  • To establish a comprehensive methodology for setting up and standardizing EV analysis using nanoscale flow cytometry.
  • To enable reliable detection, analysis, and sorting of specific EV populations.
  • To facilitate cross-platform comparison and robust quality control (QC) and quality assurance (QA) of EV measurements.

Main Methods:

  • Utilized nanoparticles of varying sizes, fluorescence intensities, and materials to establish instrument-specific distribution curves.
  • Employed these curves for flow cytometer optimization, focusing on small particle detection.
  • Developed procedures for sample preparation enabling EV recovery for downstream applications.

Main Results:

  • Demonstrated a standardized protocol for nanoscale flow cytometry setup and EV analysis.
  • Established a framework for instrument optimization and reference curve generation using controls.
  • Enabled precise detection, analysis, and sorting of specific EV populations.

Conclusions:

  • The described methodology provides a robust platform for standardized EV analysis via nanoscale flow cytometry.
  • This approach enhances the reliability of EV measurements, supporting QC/QA and inter-laboratory comparisons.
  • The protocol facilitates the recovery of EVs for subsequent functional and molecular analyses.