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Related Experiment Videos

Modelling the individual cell lag phase. Isolating single cells: protocol development.

K Francois1, F Devlieghere, A R Standaert

  • 1Laboratory of Food Microbiology and Preservation, Ghent University, Coupure Links 653, 9000 Ghent, Belgium.

Letters in Applied Microbiology
|June 14, 2003
PubMed
Summary

This study presents a new protocol for isolating single cells in microtitre plates with high certainty and yield. This method enhances the investigation of individual cell lag phases in microbial studies.

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Area of Science:

  • Microbiology
  • Cell Biology
  • Biotechnology

Background:

  • Accurate isolation of single cells is crucial for microbial studies, particularly for analyzing individual cell lag phases.
  • Existing methods often face challenges in achieving both high certainty of single-cell presence and sufficient yield.

Purpose of the Study:

  • To develop and validate a protocol for isolating single cells in microtitre plate wells.
  • To ensure high certainty of individual cell isolation and maintain a sufficient yield.
  • To provide a foundation for studying the individual cell lag phase.

Main Methods:

  • Utilized a 1/2 dilution series in microtitre plates for single-cell isolation.
  • Employed plate counting to assess seventy-two Lactococcus lactis dilution series.

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  • Validated results using a simulation model and compared the method with conventional approaches.

    • Main Results:

    • Achieved an 80% probability of obtaining a single cell per well in the final columns of the plates.
    • Reported a yield of 75 wells containing cells.
    • Simulation model confirmed the experimental findings.

    Conclusions:

    • The developed protocol effectively combines a higher probability of single-cell isolation per well with a slightly increased overall yield.
    • This method provides a valuable tool for isolating single cells, supporting research into individual cell lag phase dynamics.