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Opt Lett. 2007 Sep 1;32(17):2608-10.

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

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  • 1Department of Biochemistry and Biophysics, University of California San Francisco, 1550 Fourth Street, San Francisco, California 94158-2324, USA.


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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