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Flow Cytometry01:23

Flow Cytometry

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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An electroporation cytometry system for long-term, live cell cycle analysis.

Thomas Nesmith, Christian Vieira1, Darius G Rackus

  • 1Department of Chemistry and Biology, Toronto Metropolitan University, Toronto, Ontario M5B 2K3, Canada.

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Summary

This study introduces a novel electroporation cytometry system for real-time analysis of electric field effects on cells. The system revealed that electric pulses impact cell cycle progression in mammalian cells.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Electroporation Techniques

Background:

  • Electric fields are versatile tools in biology, utilized in methods like electroporation, gene electrotransfer (GET), electrostimulation (ES), and electrochemotherapy, each with distinct cellular outcomes.
  • While electrostimulation (ES) is known to affect cell cycle progression without pore formation, the impact of pore-forming electric fields, such as in GET, remains less explored despite clinical relevance.
  • A gap exists in commercial systems for real-time visual analysis of electric field effects on mammalian cell cultures.

Purpose of the Study:

  • To develop and validate an electroporation cytometry system enabling live cell imaging and analysis of electric field impacts.
  • To investigate the effects of pore-forming electric fields on cell cycle progression in mammalian cells.
  • To address the limitations of current commercial systems in real-time visual analysis of electroporation.

Main Methods:

  • Development of a custom electroporation chamber compatible with live cell imaging.
  • Integration of an exponential decay pulse generator for precise electrical field application.
  • Utilizing a U-2 OS cell line with a FUCCI(CA)5 cell cycle reporter for functional demonstration.
  • Application of a 180 V pulse to both unsynchronized and synchronized cell populations.

Main Results:

  • The developed electroporation cytometry system successfully facilitated live cell analysis.
  • Exposure to a 180 V electric pulse demonstrated a discernible effect on the cell cycle progression of U-2 OS cells.
  • The system's functionality was validated using a synchronized and unsynchronized cell population.

Conclusions:

  • The novel electroporation cytometry system provides a platform for investigating real-time electric field effects on cells, particularly concerning cell cycle dynamics.
  • Pore-forming electric fields, as applied in this study, influence mammalian cell cycle progression.
  • This technology has the potential to advance research in electroporation and related cellular manipulation techniques.