Outlook on optimizing ultramicroscopy imaging technique through optical characterization
- PMID: 28000307
- DOI: 10.1002/jemt.22815
Outlook on optimizing ultramicroscopy imaging technique through optical characterization
Abstract
Here, we present an optically optimized system for static ultramicroscopy imaging technique. The unit for generating an ultra-thin light sheet employs aspheric and meso-optical elements (meso-aspheric system). An analytical as well as an experimental comparison between the light sheet produced by the standard system (using a rectangular slit aperture and one cylindrical lens) and the one produced by our latest optimized system, which converts a symmetrical Gaussian beam into an ultra-thin light sheet is presented. Using the new light sheet in combination with our objective equipped with a modulator unit to compensate the refractive index mismatch between air and mediums with indices of 1.45-1.56, we present high resolution images of various biological samples that were chemically cleared using different methods. They demonstrate a marked improvement in quality, contrast and resolution.
Keywords: Gaussian beam; light sheet microscopy; optical parameters characterization; uniform beam profile.
© 2016 Wiley Periodicals, Inc.
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