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Related Experiment Videos

Sizing biological samples by photosedimentation techniques.

A P Middelberg1, I D Bogle, M A Snoswell

  • 1Department of Chemical Engineering, University of Adelaide, Australia.

Biotechnology Progress
|July 1, 1990
PubMed
Summary
This summary is machine-generated.

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The Joyce-Loebl disk centrifuge accurately sizes Escherichia coli cells and particles when using a glycerol-water buffer. Ethanol affects cell size, and diffusion impacts sizing of very small particles below 0.01 microns.

Area of Science:

  • Biophysical characterization
  • Particle sizing technology
  • Microbiology

Background:

  • Accurate sizing of biological particles like Escherichia coli (E. coli) is crucial for various applications.
  • The Joyce-Loebl disk centrifuge is a tool for particle size analysis, but its performance with biological samples requires evaluation.
  • Standardization of experimental conditions is necessary for reliable results.

Purpose of the Study:

  • To evaluate the performance of the Joyce-Loebl disk centrifuge for sizing E. coli, protein inclusion bodies, and cell debris.
  • To define optimal spin fluid conditions for accurate bacterial sizing.
  • To analyze factors affecting measurement accuracy, including particle-particle interactions and diffusion.

Main Methods:

  • Performance evaluation of the Joyce-Loebl disk centrifuge.

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  • Determination of optimal spin-buffer fluid combinations (e.g., glycerol-water).
  • Analysis of particle diffusion and sedimentation within the spin fluid.

    • Main Results:

    • A density gradient extending throughout the spin fluid is essential for accurate sizing.
    • Ethanol exposure reduces the Stokes diameter of E. coli cells, favoring glycerol-water buffers for bacterial sizing.
    • Baseline drift and diffusion effects were analyzed, with diffusion being significant for particles near the 0.01-micron limit.

    Conclusions:

    • The Joyce-Loebl disk centrifuge can effectively size E. coli and other biological particles under defined conditions.
    • Glycerol-water spin fluid is recommended for sizing bacteria to avoid ethanol-induced size reduction.
    • Careful consideration of diffusion is needed for sizing sub-0.01 micron particles.