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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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BG-flow, a new flow cytometry tool for G-quadruplex quantification in fixed cells.

Alessio De Magis1, Melanie Kastl1, Peter Brossart1

  • 1Department of Oncology, Hematology and Rheumatology, University Hospital Bonn, Venusberg-Campus 1, 53127, Bonn, Germany.

BMC Biology
|March 12, 2021
PubMed
Summary

A new flow cytometry method (BG-flow) quantifies G-quadruplexes (G4s) in cells. This technique offers a faster way to study G4 structures and their roles in cellular homeostasis.

Keywords:
Flow cytometryG-quadruplexSecondary DNA structures

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • G-quadruplexes (G4s) are non-canonical nucleic acid structures with established in vitro properties but debated in vivo functions.
  • G4s are found at specific genomic locations and implicated in cellular homeostasis.
  • Current methods for detecting G4s in vivo, like microscopy and sequencing, are time-consuming.

Purpose of the Study:

  • To develop and validate a robust, time-efficient biochemical method for quantifying G-quadruplex structures in living cells.
  • To enable deeper understanding of G4 structure functions and formation triggers in vivo.

Main Methods:

  • Development of a novel protocol, BG-flow, utilizing the G4-specific antibody BG4 for G4 quantification via flow cytometry.
  • Validation of BG-flow in various cell types, including human cultured cells (Hela, THP-1), primary human blood cells (PBMCs), and mouse macrophages.

Main Results:

  • BG-flow successfully quantifies G4 levels across diverse cell types from mice to humans.
  • The method detected changes in G4 levels during the cell cycle in MCF-7 cells.
  • BG-flow validated previously observed changes in G4 levels upon treatment with pyridostatin (PDS).

Conclusions:

  • BG-flow provides mechanistic proof of concept across species and cell types.
  • BG-flow can be combined with cell surface markers for G4 analysis in specific cell subpopulations.
  • This method will aid research into G4 structures as potential diagnostic tools.