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

PLoS One. 2017 Jul 11;12(7):e0180309. doi: 10.1371/journal.pone.0180309. eCollection 2017.

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.

MeSH terms

  • Algorithms
  • Animals
  • Microscopy / methods*
  • Software*
  • Tomography, Optical / methods*
  • Zebrafish

Grants and funding

The authors gratefully acknowledge funding from the UK Biotechnology and Biological Sciences Research Council (BBSRC BB/L018039/1). Thomas Watson and Samuel Davis acknowledge Ph.D. studentships from the UK Engineering and Physical Sciences Research Council (EPSRC) and Natalie Andrews an EPSRC Doctoral Prize Fellowship.