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. 2018 Sep;81(9):929-935.
doi: 10.1002/jemt.22815. Epub 2016 Dec 20.

Outlook on optimizing ultramicroscopy imaging technique through optical characterization

Saiedeh Saghafi  1   2 , Klaus Becker  1   2 , Christian Hahn  1   2 , Marko Pende  1   2 , Nina Jährling  1   2 , Martina Wanis  1   2 , Inna Sabdyusheva-Litschauer  1   2 , Massih Foroughipour  1   2 , Axel Niendorf  3 , Hans-Ulrich Dodt  1   2
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Outlook on optimizing ultramicroscopy imaging technique through optical characterization

Saiedeh Saghafi et al. Microsc Res Tech. 2018 Sep.

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.

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