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Imaging Spectrometers for Fluorescence and Raman Microscopy: Acousto-Optic and Liquid Crystal Tunable Filters

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Abstract:

Acousto-optic tunable filters (AOTF) and liquid crystal tunable filters (LCTF) are evaluated for their suitability as fluorescence microscopy imaging spectrometers. AOTFs are solid-state birefringent crystals that provide an electronically tunable spectral notch passband in response to an applied acoustic field. LCTFs also provide a notch passband that can be controlled by incorporating liquid crystal waveplate retarders within a Lyot birefringent filter. In this paper, spectroscopic performance and imaging quality are contrasted by evaluation of model systems. Studies include transmission imaging of standard resolution targets, multispectral fluorescence emission imaging of tagged polystyrene microspheres, and immunofluorescence imaging of neurotransmitters within rat-brainstem thin sections. In addition, the first use of LCTFs for Raman microscopy is demonstrated. Raman microscopy is a noninvasive spectral imaging technique that can provide chemically significant image contrast complementary to fluorescence microscopy without the use of stains or tags.

Keywords: Acousto-optic tunable filter (AOTF); Immunofluorescence microscopy; Liquid crystal tunable filter (LCTF); Lyot filter; Microspheres; Neurotransmitters; Polystyrene; Raman microscopy; Reticulo-spinal neurons; Serotonin (5HT); Spectral imaging

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/0003702944029820

Affiliations: 1: Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260 2: Cambridge Research & Instrumentation Inc., 21 Erie Street, Cambridge, Massachusetts 02139

Publication date: July 1, 1994

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