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Related Experiment Videos

High-pressure fluorescence correlation spectroscopy.

Joachim D Müller1, Enrico Gratton

  • 1Laboratory for Fluorescence Dynamics, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.

Biophysical Journal
|September 26, 2003
PubMed
Summary
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We developed a novel high-pressure cell using a fused silica microcapillary for fluorescence correlation spectroscopy (FCS). This setup enables fluorescence fluctuation spectroscopy under pressures up to 300 MPa.

Area of Science:

  • Analytical Chemistry
  • Physical Chemistry
  • Spectroscopy

Background:

  • Fluorescence correlation spectroscopy (FCS) is a powerful technique for studying molecular dynamics.
  • High-pressure studies are crucial for understanding molecular behavior in condensed phases.
  • Existing high-pressure cells often have limitations for optical measurements.

Purpose of the Study:

  • To demonstrate the suitability of a novel fused silica microcapillary-based high-pressure cell for FCS.
  • To characterize and mitigate optical distortions caused by the capillary's geometry.
  • To report the first experimental realization of fluorescence fluctuation spectroscopy under high pressure.

Main Methods:

  • Development of a high-pressure cell using a single fused silica microcapillary.

Related Experiment Videos

  • Characterization of excitation light refraction and point spread function distortions using FCS and photon-counting histogram (PCH) analysis.
  • Optimization of experimental position within the capillary for accurate measurements.
  • Measurement of autocorrelation functions and photon count distributions from 0 to 300 MPa using a fluorescent dye model system.

    • Main Results:

    • Identified an optimal position within the capillary where standard FCS/PCH equations apply.
    • Successfully performed fluorescence fluctuation spectroscopy at pressures up to 300 MPa.
    • Observed a small pressure dependence (approx. 10%) in fluctuation amplitude and diffusion coefficient.
    • Attributed observed parameter changes to pressure-induced alterations in viscosity and density of the aqueous medium.

    Conclusions:

    • The novel fused silica microcapillary cell is suitable for high-pressure FCS.
    • The developed method allows for the study of molecular dynamics under high-pressure conditions.
    • The observed pressure dependence provides insights into the physical properties of the medium.