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A combined light sheet fluorescence and differential interference contrast microscope for live imaging of multicellular specimens
Author(s) -
BAKER R.P.,
TAORMINA M.J.,
JEMIELITA M.,
PARTHASARATHY R.
Publication year - 2015
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12220
Subject(s) - differential interference contrast microscopy , light sheet fluorescence microscopy , microscopy , fluorescence microscope , interference microscopy , microscope , optical microscope , optical sectioning , bright field microscopy , optics , two photon excitation microscopy , contrast (vision) , materials science , fluorescence , physics , scanning electron microscope
Summary We describe a microscope capable of both light sheet fluorescence microscopy and differential interference contrast microscopy (DICM). The two imaging modes, which to the best of our knowledge have not previously been combined, are complementary: light sheet fluorescence microscopy provides three‐dimensional imaging of fluorescently labelled components of multicellular systems with high speed, large fields of view, and low phototoxicity, whereas differential interference contrast microscopy reveals the unlabelled neighbourhood of tissues, organs, and other structures with high contrast and inherent optical sectioning. Use of a single Nomarski prism for differential interference contrast microscopy and a shared detection path for both imaging modes enables simple integration of the two techniques in one custom microscope. We provide several examples of the utility of the resulting instrument, focusing especially on the digestive tract of the larval zebrafish, revealing in this complex and heterogeneous environment anatomical features, the behaviour of commensal microbes, immune cell motions, and more.