ZFIN ID: ZDB-PUB-170713-1
OPTiM: Optical projection tomography integrated microscope using open-source hardware and software
Watson, T., Andrews, N., Davis, S., Bugeon, L., Dallman, M.D., McGinty, J.
Date: 2017
Source: PLoS One   12: e0180309 (Journal)
Registered Authors: Andrews, Natalie, Bugeon, Laurence, Dallman, Maggie
Keywords: Zebrafish, Optical lenses, Fluorescence imaging, Inverted microscopy, Light microscopy, Relays, Scanning confocal microscopy, Focal planes
MeSH Terms:
  • Algorithms
  • Animals
  • Microscopy/methods*
  • Software*
  • Tomography, Optical/methods*
  • Zebrafish
PubMed: 28700724 Full text @ PLoS One
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ABSTRACT
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.
ADDITIONAL INFORMATION