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Related Experiment Videos

Application of binary buffer systems to free flow cell electrophoresis.

G Weber1, D Grimm, J Bauer

  • 1Kirchheim, Germany.

Electrophoresis
|February 16, 2000
PubMed
Summary

Free flow electrophoresis (FFE) now processes sensitive cells using a novel, non-toxic buffer. This advancement allows for precise electrophoretic behavior determination of human cells, maintaining viability for reculturing.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Analytical Chemistry

Background:

  • Free flow electrophoresis (FFE) is a powerful separation technique.
  • Processing sensitive cells with FFE has been challenging due to medium requirements.
  • Existing methods may compromise cell viability or cause overheating.

Purpose of the Study:

  • To adapt Free Flow Electrophoresis (FFE) for sensitive cell processing.
  • To develop a new electrophoresis medium that supports cell viability.
  • To accurately determine the electrophoretic behavior of human cells using FFE.

Main Methods:

  • Adjusting chamber medium pH to 7.35 using EPPS and BISTRIS buffers.
  • Preparing a new cell electrophoresis separation medium with 30 mM NaCl and optional 1 mM CaCl2.

Related Experiment Videos

  • Utilizing the Octopus preparative FFE device for cell separation and analysis.
  • Assessing cell viability and reculturing potential post-electrophoresis.

    • Main Results:

    • EPPS and BISTRIS buffers were found to be non-toxic to human smooth muscle and thyroid cells.
    • The new medium slightly enhanced electrical conductivity, preventing overheating.
    • Human smooth muscle and thyroid carcinoma cells remained viable single cells for over 120 minutes.
    • Electrophoresed cells migrated individually without clotting, enabling exact behavior determination and successful reculturing.

    Conclusions:

    • Free flow electrophoresis (FFE) is now applicable to sensitive cell types.
    • The developed non-toxic medium and optimized conditions ensure cell viability and accurate electrophoretic analysis.
    • This method expands FFE's utility in biological and medical research for cell characterization.