title: Mirrored pyramidal wells for simultaneous multiple vantage point microscopy.
abstract: 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.
authors.affiliation: Department of Biological Sciences, Vanderbilt University, VU Station B 351807, Nashville, TN 37235-1807, USA.
authors.forename: K T
authors.initials: KT
authors.lastname: Seale
authors.forename: R S
authors.initials: RS
authors.lastname: Reiserer
authors.forename: D A
authors.initials: DA
authors.lastname: Markov
authors.forename: I A
authors.initials: IA
authors.lastname: Ges
authors.forename: C
authors.initials: C
authors.lastname: Wright
authors.forename: C
authors.initials: C
authors.lastname: Janetopoulos
authors.forename: J P
authors.initials: JP
authors.lastname: Wikswo
mesh_terms: D000818:Animals; D004023:Dictyostelium / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional / Q000379:methods; D008853:Microscopy / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D015388:Organelles / Q000648:ultrastructure; D013048:Specimen Handling / Q000379:methods*