title: Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM).
abstract: 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.
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, 1550 Fourth Street, San Francisco, California 94158-2324, USA. jan.huisken@ucsf.edu
authors.forename: Jan
authors.initials: J
authors.lastname: Huisken
authors.forename: Didier Y R
authors.initials: DY
authors.lastname: Stainier
mesh_terms: D000042:Absorption; D000818:Animals; D004625:Embryo, Nonmammalian / Q000473:pathology; D004867:Equipment Design; D007091:Image Processing, Computer-Assisted; D008027:Light; D008853:Microscopy / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D055095:Optics and Photonics; D012542:Scattering, Radiation; D012984:Software; D013997:Time Factors; D015027:Zebrafish