Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples

Nat Methods. 2008 Jun;5(6):527-9. doi: 10.1038/nmeth.1211. Epub 2008 May 11.

Abstract

Imaging volumes as thick as whole cells at three-dimensional (3D) super-resolution is required to reveal unknown features of cellular organization. We report a light microscope that generates images with translationally invariant 30 x 30 x 75 nm resolution over a depth of several micrometers. This method, named biplane (BP) FPALM, combines a double-plane detection scheme with fluorescence photoactivation localization microscopy (FPALM) enabling 3D sub-diffraction resolution without compromising speed or sensitivity.

Publication types

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

MeSH terms

  • Biophysics / economics
  • Biophysics / instrumentation
  • Biophysics / methods*
  • Fluorescein / pharmacology
  • Fluorescent Dyes / pharmacology
  • Image Enhancement
  • Image Interpretation, Computer-Assisted / methods
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods*
  • Lasers
  • Light
  • Microscopy / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Software

Substances

  • Fluorescent Dyes
  • Fluorescein