title: Mapping optical path length and image enhancement using quantitative orientation-independent differential interference contrast microscopy.
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.
authors.affiliation: Marine Biological Laboratory, 7 MBL Street, Woods Hole, Massachusetts 02543, Unites States.
authors.forename: Michael
authors.initials: M
authors.lastname: Shribak
authors.affiliation: Nontrivialzeros Research, 22 Mitchell Street, Putney, New South Wales 2112, Australia.
authors.forename: Kieran G
authors.initials: KG
authors.lastname: Larkin
authors.affiliation: KB Imaging Solutions, 3849 Val Verde Road, Loomis, California 95650, Unites States.
authors.forename: David
authors.initials: D
authors.lastname: Biggs
mesh_terms: D007089:Image Enhancement / Q000379:methods*; D008853:Microscopy / Q000295:instrumentation; D008857:Microscopy, Interference / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*