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Proceedings Paper

Adaptive optimisation of illumination beam profiles in fluorescence microscopy
Author(s): T. J. Mitchell; C. D. Saunter; W. O'Nions; J. M. Girkin; Gordon D. Love
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Paper Abstract

Wide-field fluorescence microscope techniques such as single/selective plane illumination microscope (SPIM) are typically configured to image large regions of a sample at once. Here the illumination beam provides uniform excitation of several biological features across the region, `sliced' to a thickness of between 5-10 microns. In this paper we propose a simple alteration to the optical configuration of a SPIM by switching the light-sheet- forming cylindrical lens with a spatial light modulator. This has the potential to adaptively reconfigure the light sheet geometry to improve the optical sectioning of specific biological features, rather than the thicker sectioning of several features at once across a larger observation field-of-view. We present a prototype version of such a system, referred to as an Adaptive-SPIM (A-SPIM) system. We then suggest that the direct recording of illumination beam shapes within the working microscope system can better facilitate the analysis and subsequent re-configuration of the illumination beam to a specific geometry, and summarise the design and operation of a device that we have developed specifically for this purpose. We finally present reconstructed quantitative three dimensional flux maps of illumination beams from three microscope configurations taken using this miniature high-dynamic range beam profiling device, comparing the beam geometry of a regular SPIM system with our prototype A-SPIM system, and suggesting future improvements.

Paper Details

Date Published: 10 March 2015
PDF: 6 pages
Proc. SPIE 9335, Adaptive Optics and Wavefront Control for Biological Systems, 93350B (10 March 2015); doi: 10.1117/12.2080310
Show Author Affiliations
T. J. Mitchell, Durham Univ. (United Kingdom)
C. D. Saunter, Durham Univ. (United Kingdom)
W. O'Nions, Durham Univ. (United Kingdom)
J. M. Girkin, Durham Univ. (United Kingdom)
Gordon D. Love, Durham Univ. (United Kingdom)

Published in SPIE Proceedings Vol. 9335:
Adaptive Optics and Wavefront Control for Biological Systems
Thomas G. Bifano; Joel Kubby; Sylvain Gigan, Editor(s)

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