Mapping optical path length and image enhancement using quantitative orientation-independent differential interference contrast microscopy

Michael Shribak; Kieran G. Larkin; David Biggs

doi:10.1117/1.JBO.22.1.016006

6 January 2017 Mapping optical path length and image enhancement using quantitative orientation-independent differential interference contrast microscopy

Michael Shribak, Kieran G. Larkin, David Biggs

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Journal of Biomedical Optics, Vol. 22, Issue 1, 016006 (January 2017). https://doi.org/10.1117/1.JBO.22.1.016006

Abstract

We describe the principles of using orientation-independent differential interference contrast (OI-DIC) microscopy for mapping optical path length (OPL). Computation of the scalar two-dimensional OPL map is based on an experimentally received map of the OPL gradient vector field. Two methods of contrast enhancement for the OPL image, which reveal hardly visible structures and organelles, are presented. The results obtained can be used for reconstruction of a volume image. We have confirmed that a standard research grade light microscope equipped with the OI-DIC and 100×/1.3 NA objective lens, which was not specially selected for minimum wavefront and polarization aberrations, provides OPL noise level of ∼0.5 nm and lateral resolution if ∼300 nm at a wavelength of 546 nm. The new technology is the next step in the development of the DIC microscopy. It can replace standard DIC prisms on existing commercial microscope systems without modification. This will allow biological researchers that already have microscopy setups to expand the performance of their systems.

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Michael Shribak, Kieran G. Larkin, and David Biggs "Mapping optical path length and image enhancement using quantitative orientation-independent differential interference contrast microscopy," Journal of Biomedical Optics 22(1), 016006 (6 January 2017). https://doi.org/10.1117/1.JBO.22.1.016006

Received: 24 September 2016; Accepted: 12 December 2016; Published: 6 January 2017

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