OPTiM: Optical projection tomography integrated microscope using open-source hardware and software

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 sta...

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Veröffentlicht in:PloS one 2017-07, Vol.12 (7), p.e0180309-e0180309
Hauptverfasser: Watson, Thomas, Andrews, Natalie, Davis, Samuel, Bugeon, Laurence, Dallman, Margaret D, McGinty, James
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Sprache:eng
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Zusammenfassung: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0180309