Fluorescence imaging with one nanometer accuracy: application to molecular motors

Acc Chem Res. 2005 Jul;38(7):574-82. doi: 10.1021/ar040136s.

Abstract

We introduce the technique of FIONA, fluorescence imaging with one nanometer accuracy. This is a fluorescence technique that is able to localize the position of a single dye within approximately 1 nm in the x-y plane. It is done simply by taking the point spread function of a single fluorophore excited with wide field illumination and locating the center of the fluorescent spot by a two-dimensional Gaussian fit. We motivate the development of FIONA by unraveling the walking mechanism of the molecular motors myosin V, myosin VI, and kinesin. We find that they all walk in a hand-over-hand fashion.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biophysical Phenomena
  • Biophysics
  • Carbocyanines
  • Fluorescent Dyes
  • Green Fluorescent Proteins
  • Kinesin / chemistry*
  • Kinesin / metabolism
  • Microscopy, Fluorescence / methods*
  • Models, Molecular
  • Molecular Motor Proteins / chemistry*
  • Molecular Motor Proteins / metabolism
  • Myosin Heavy Chains / chemistry*
  • Myosin Heavy Chains / metabolism
  • Myosin Type V / chemistry*
  • Myosin Type V / metabolism
  • Nanotechnology
  • Rhodamines

Substances

  • Carbocyanines
  • Fluorescent Dyes
  • Molecular Motor Proteins
  • Rhodamines
  • cyanine dye 3
  • myosin VI
  • Green Fluorescent Proteins
  • Myosin Type V
  • Myosin Heavy Chains
  • Kinesin