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Fluorescence correlation spectroscopy: diagnostics for sparse molecules

S Maiti1, U Haupts, W W Webb

  • 1Applied Physics, Cornell University, Ithaca, NY 14853, USA.

Proceedings of the National Academy of Sciences of the United States of America
|October 29, 1997
PubMed
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Molecular fluorescence enables detection of sparse molecules. Correlation spectroscopy analyzes these signals for quantitative insights into concentration, mobility, and photophysics, even for single molecules.

Area of Science:

  • Analytical Chemistry
  • Biophysics
  • Physical Chemistry

Background:

  • Molecular fluorescence allows sensitive detection of molecules.
  • Sparse molecular detection is crucial for various scientific disciplines.
  • Existing methods may have limitations in sensitivity or scope.

Purpose of the Study:

  • To highlight the capabilities of molecular fluorescence and correlation spectroscopy.
  • To demonstrate the application of these techniques for analyzing molecules at low concentrations.
  • To showcase the potential for studying individual molecules.

Main Methods:

  • Utilizing the robust glow of molecular fluorescence for detection.
  • Employing correlation spectroscopy, a statistical-physics-based tool.

Related Experiment Videos

  • Analyzing spontaneously fluctuating fluorescence signals from molecular ensembles.

    • Main Results:

    • Sparse molecules are detectable and amenable to analysis.
    • Quantitative information on concentration, mobility, chemistry, and photophysics can be obtained.
    • Analysis is feasible at concentrations less than one nanomolar in solution and less than one macromolecule per square micrometer on cell surfaces.

    Conclusions:

    • Molecular fluorescence and correlation spectroscopy offer powerful analytical capabilities.
    • These techniques are effective for studying molecules at minuscule concentrations.
    • The detection, localization, and examination of individual molecules are becoming increasingly common.