4Pi Microscopy

Methods Mol Biol. 2013:950:27-41. doi: 10.1007/978-1-62703-137-0_3.

Abstract

Optical microscopy has become a key technology in the life sciences today. Its noninvasive nature provides access to the interior of intact and even living cells, where specific molecules can be precisely localized by fluorescent tagging. However, the attainable 3D resolution of an optical microscope has long been hampered by a comparatively poor resolution along the optic axis. By coherent focusing through two objective lenses, 4Pi microscopy improves the axial resolution by three- to fivefold. This primer is intended as a starting point for the design and operation of a 4Pi microscope of type A.

MeSH terms

  • Cell Nucleus Structures / metabolism
  • Golgi Apparatus / metabolism
  • Image Processing, Computer-Assisted
  • Microscopy / methods*
  • Optical Phenomena*
  • Sulfhydryl Compounds / chemistry

Substances

  • Sulfhydryl Compounds
  • 2,2'-thiodiethanol