Fast molecular tracking maps nanoscale dynamics of plasma membrane lipids

Proc Natl Acad Sci U S A. 2010 Apr 13;107(15):6829-34. doi: 10.1073/pnas.0912894107. Epub 2010 Mar 29.

Abstract

We describe an optical method capable of tracking a single fluorescent molecule with a flexible choice of high spatial accuracy (approximately 10-20 nm standard deviation or approximately 20-40 nm full-width-at-half-maximum) and temporal resolution (< 1 ms). The fluorescence signal during individual passages of fluorescent molecules through a spot of excitation light allows the sequential localization and thus spatio-temporal tracking of the molecule if its fluorescence is collected on at least three separate point detectors arranged in close proximity. We show two-dimensional trajectories of individual, small organic dye labeled lipids diffusing in the plasma membrane of living cells and directly observe transient events of trapping on < 20 nm spatial scales. The trapping is cholesterol-assisted and much more pronounced for a sphingo- than for a phosphoglycero-lipid, with average trapping times of approximately 15 ms and < 4 ms, respectively. The results support previous STED nanoscopy measurements and suggest that, at least for nontreated cells, the transient interaction of a single lipid is confined to macromolecular dimensions. Our experimental approach demonstrates that fast molecular movements can be tracked with minimal invasion, which can reveal new important details of cellular nano-organization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Diffusion
  • Kidney / metabolism
  • Membrane Lipids / metabolism*
  • Membrane Microdomains / chemistry*
  • Membrane Microdomains / metabolism
  • Microscopy, Confocal / methods
  • Potoroidae
  • Probability
  • Reproducibility of Results
  • Signal Processing, Computer-Assisted
  • Time Factors

Substances

  • Membrane Lipids