Open AccessOPTiM: Optical projection tomography integrated microscope using open-source hardware and software
Author(s) -
Thomas J. Watson,
Natalie Andrews,
Samuel P. X. Davis,
Laurence Bugeon,
Margaret D. Dallman,
James McGinty
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0180309
Subject(s) - microscope , optics , lens (geology) , stepper , computer science , magnification , tomography , aperture (computer memory) , projection (relational algebra) , field of view , software , physics , acoustics , algorithm , programming language
We describe the implementation of an OPT plate to perform optical projection tomography (OPT) on a commercial wide-field inverted microscope, using our open-source hardware and software. The OPT plate includes a tilt adjustment for alignment and a stepper motor for sample rotation as required by standard projection tomography. Depending on magnification requirements, three methods of performing OPT are detailed using this adaptor plate: a conventional direct OPT method requiring only the addition of a limiting aperture behind the objective lens; an external optical-relay method allowing conventional OPT to be performed at magnifications >4x; a remote focal scanning and region-of-interest method for improved spatial resolution OPT (up to ~1.6 μm). All three methods use the microscope’s existing incoherent light source (i.e. arc-lamp) and all of its inherent functionality is maintained for day-to-day use. OPT acquisitions are performed on in vivo zebrafish embryos to demonstrate the implementations’ viability.