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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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Large Extracellular Vesicles Can be Characterised by Multiplex Labelling Using Imaging Flow Cytometry.

Suzanne M Johnson1, Antonia Banyard2, Christopher Smith1

  • 1Children's Cancer Group, Division of Cancer Sciences, School of Medical Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oglesby Cancer Research Building, Manchester Academic Health Science Centre, Manchester Cancer Research Centre, Manchester M20 4GJ, UK.

International Journal of Molecular Sciences
|November 21, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method to isolate and analyze large extracellular vesicles (EVs) from medulloblastoma cells. This technique allows for detailed characterization of individual EVs, revealing potential biomarkers for disease monitoring.

Keywords:
biomarker reservoirscancer diagnosticsdisease monitoringextracellular vesiclesimaging flow cytometrylarge EVs

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Area of Science:

  • Extracellular vesicle (EV) research
  • Biotechnology
  • Cancer biology

Background:

  • Extracellular vesicles (EVs) are diverse in size and content.
  • Current isolation methods often exclude larger EVs.
  • Large EVs may hold significant diagnostic potential.

Purpose of the Study:

  • To develop a standardized method for isolating and characterizing large EVs from medulloblastoma cells.
  • To utilize imaging flow cytometry for detailed analysis of individual large EVs.
  • To investigate EV marker expression in relation to EV size.

Main Methods:

  • Isolation of large EVs from UW228-2 medulloblastoma cell line.
  • Characterization using fluorescent microscopy, TEM, and TRPS.
  • Imaging flow cytometry with fluorescent membrane labeling and stringent gating.
  • Assessment of EV markers CD63, CD9, and LAMP1.

Main Results:

  • UW228-2 cells release large EVs up to 6 µm.
  • Imaging flow cytometry provides robust analysis of large EVs, including diameter.
  • A correlation exists between increasing EV size and marker co-expression.

Conclusions:

  • A novel labeling and gating strategy enables individual EV marker analysis.
  • This method is effective for heterogeneous EV populations.
  • Large EVs show promise as biomarkers for diagnostic screening and disease monitoring.