Ultra-High Resolution 3D Imaging of Whole Cells

Cell. 2016 Aug 11;166(4):1028-1040. doi: 10.1016/j.cell.2016.06.016. Epub 2016 Jul 7.

Abstract

Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples. Here, we present the development of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), an optical nanoscope that allows imaging of three-dimensional (3D) structures at 10- to 20-nm resolution throughout entire mammalian cells. We demonstrate the wide applicability of W-4PiSMSN across diverse research fields by imaging complex molecular architectures ranging from bacteriophages to nuclear pores, cilia, and synaptonemal complexes in large 3D cellular volumes.

Publication types

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

MeSH terms

  • Animals
  • Bacteriophages / ultrastructure
  • COP-Coated Vesicles / ultrastructure
  • Cytological Techniques / instrumentation
  • Cytological Techniques / methods*
  • Golgi Apparatus / ultrastructure
  • Male
  • Mice
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Single Molecule Imaging / instrumentation
  • Single Molecule Imaging / methods*
  • Spermatocytes / ultrastructure
  • Synaptonemal Complex / ultrastructure