Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM)

Opt Lett. 2007 Sep 1;32(17):2608-10. doi: 10.1364/ol.32.002608.


Multidirectional selective plane illumination microscopy (mSPIM) reduces absorption and scattering artifacts and provides an evenly illuminated focal plane. mSPIM solves two common problems in light-sheet-based imaging techniques: The shadowing in the excitation path due to absorption in the specimen is eliminated by pivoting the light sheet; the spread of the light sheet by scattering in the sample is compensated by illuminating the sample consecutively from opposing directions. The resulting two images are computationally fused yielding a superior image. The effective light sheet is thinner, and the axial resolution is increased by square root 2 over single-directional SPIM. The multidirectional illumination proves essential in biological specimens such as millimeter-sized embryos. The performance of mSPIM is demonstrated by the imaging of live zebrafish embryos.

Publication types

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

MeSH terms

  • Absorption
  • Animals
  • Embryo, Nonmammalian / pathology
  • Equipment Design
  • Image Processing, Computer-Assisted
  • Light
  • Microscopy / methods*
  • Microscopy, Fluorescence / methods*
  • Optics and Photonics
  • Scattering, Radiation
  • Software
  • Time Factors
  • Zebrafish