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

J Huisken, DYR Stainier - Optics letters, 2007 - opg.optica.org
Optics letters, 2007opg.optica.org
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 …
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.
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