Interferometric fluorescence cross correlation spectroscopy

PLoS One. 2019 Dec 18;14(12):e0225797. doi: 10.1371/journal.pone.0225797. eCollection 2019.

Abstract

Measuring transport properties like diffusion and directional flow is essential for understanding dynamics within heterogeneous systems including living cells and novel materials. Fluorescent molecules traveling within these inhomogeneous environments under the forces of Brownian motion and flow exhibit fluctuations in their concentration, which are directly linked to the transport properties. We present a method utilizing single photon interference and fluorescence correlation spectroscopy (FCS) to simultaneously measure transport of fluorescent molecules within aqueous samples. Our method, within seconds, measures transport in thousands of homogenous voxels (100 nm)3 and under certain conditions, eliminates photo-physical artifacts associated with blinking of fluorescent molecules. A comprehensive theoretical framework is presented and validated by measuring transport of quantum dots, associated with VSV-G receptor along cellular membranes as well as within viscous gels.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Membrane / chemistry*
  • Diffusion
  • HeLa Cells
  • Humans
  • Interferometry / methods
  • Light
  • Membrane Glycoproteins / chemistry*
  • Motion
  • Quantum Dots / chemistry*
  • Spectrometry, Fluorescence / methods*
  • Viral Envelope Proteins / chemistry*

Substances

  • G protein, vesicular stomatitis virus
  • Membrane Glycoproteins
  • Viral Envelope Proteins