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Single-cell quantification of molecules and rates using open-source microscope-based cytometry.

Andrew Gordon1, Alejandro Colman-Lerner, Tina E Chin

  • 1The Molecular Sciences Institute, 2168 Shattuck Avenue, Berkeley, California 94704, USA.

Nature Methods
|January 24, 2007
PubMed
Summary
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Researchers developed sensitive microscope methods to measure molecules in individual yeast cells. These techniques track molecular changes over time, complementing flow cytometry for quantitative cell biology studies.

Area of Science:

  • Cell biology
  • Quantitative physiology
  • Microscopy and imaging

Background:

  • Microscope-based cytometry enables high-throughput cellular studies.
  • Measuring molecular dynamics in individual cells is crucial for understanding cellular processes.

Purpose of the Study:

  • To present refined, sensitive methods for measuring individual Saccharomyces cerevisiae cells over time.
  • To enable quantitative analysis of molecular dynamics in living yeast cells.

Main Methods:

  • Development of simple 'wet' laboratory methods and microscopy procedures.
  • Integration of open-source software tools and statistical routines.
  • Achieving high sensitivity for detecting <350 fluorescent protein molecules per cell.

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Main Results:

  • Enabled new protocols for measuring maturation and degradation rates of molecular species (GFP derivative, native mRNA).
  • Demonstrated sensitivity and accuracy in tracking molecular changes in individual, unperturbed yeast cells.
  • Provided a method for calculating rates of molecular species maturation and degradation.

Conclusions:

  • The developed microscope methods offer high sensitivity and accuracy for tracking cellular changes over time.
  • These methods can complement flow cytometry for quantitative physiology studies.
  • The approach facilitates the study of molecular dynamics in single, living cells.