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Mirrored pyramidal wells for simultaneous multiple vantage point microscopy
Author(s) -
SEALE K.T.,
REISERER R.S.,
MARKOV D.A.,
GES I.A.,
WRIGHT C.,
JANETOPOULOS C.,
WIKSWO J.P.
Publication year - 2008
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/j.1365-2818.2008.02110.x
Subject(s) - microscopy , optics , fluorescence , dictyostelium discoideum , materials science , chemistry , physics , biochemistry , gene
Summary We report a novel method for obtaining simultaneous images from multiple vantage points of a microscopic specimen using size‐matched microscopic mirrors created from anisotropically etched silicon. The resulting pyramidal wells enable bright‐field and fluorescent side‐view images, and when combined with z ‐sectioning, provide additional information for 3D reconstructions of the specimen. We have demonstrated the 3D localization and tracking over time of the centrosome of a live Dictyostelium discoideum. The simultaneous acquisition of images from multiple perspectives also provides a five‐fold increase in the theoretical collection efficiency of emitted photons, a property which may be useful for low‐light imaging modalities such as bioluminescence, or low abundance surface‐marker labelling.