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Related Experiment Videos

Disintegration of isolated cells.

B M Löffler1, M Schmidt, H Kunze

  • 1Max-Planck-Institut für Experimentelle Medizin, Göttingen, Federal Republic of Germany.

Analytical Biochemistry
|June 1, 1987
PubMed
Summary

A new device efficiently disrupts cell membranes using controlled pressure extrusion. This method preserves delicate subcellular components like lysosomes and peroxisomes in various cell types.

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Biochemistry

Background:

  • Efficient cell lysis is crucial for subcellular component isolation.
  • Preserving fragile organelles during cell disruption remains a challenge.
  • Existing methods may damage or alter cellular structures.

Purpose of the Study:

  • To develop a simple and effective device for cell disintegration.
  • To optimize conditions for efficient plasma membrane disruption.
  • To ensure the integrity of sensitive subcellular components, including lysosomes and peroxisomes.

Main Methods:

  • A novel device extrudes cell suspensions through a small, interchangeable orifice.
  • Controlled pressure is applied during the extrusion process.
  • The orifice diameter is adjusted based on the specific cell type.

Main Results:

  • The device achieved efficient disintegration of isolated rat hepatocytes and rat Leydig cells.
  • Plasma membranes were effectively disrupted under the tested conditions.
  • Fragile subcellular components, specifically lysosomes and peroxisomes, were well-preserved.

Conclusions:

  • The developed device offers a simple yet effective method for cell lysis.
  • It provides optimal conditions for disrupting plasma membranes while maintaining organelle integrity.
  • This technique is suitable for isolating intact subcellular components from various cell types.

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