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Journal of Biomedical Optics

Thin-film tunable filters for hyperspectral fluorescence microscopy
Author(s): Peter F. Favreau; Clarissa Hernandez; Ashley Stringfellow Lindsey; Diego F. Alvarez; Thomas C. Rich; Prashant Prabhat; Silas J. Leavesley
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Paper Abstract

Hyperspectral imaging is a powerful tool that acquires data from many spectral bands, forming a contiguous spectrum. Hyperspectral imaging was originally developed for remote sensing applications; however, hyperspectral techniques have since been applied to biological fluorescence imaging applications, such as fluorescence microscopy and small animal fluorescence imaging. The spectral filtering method largely determines the sensitivity and specificity of any hyperspectral imaging system. There are several types of spectral filtering hardware available for microscopy systems, most commonly acousto-optic tunable filters (AOTFs) and liquid crystal tunable filters (LCTFs). These filtering technologies have advantages and disadvantages. Here, we present a novel tunable filter for hyperspectral imaging—the thin-film tunable filter (TFTF). The TFTF presents several advantages over AOTFs and LCTFs, most notably, a high percentage transmission and a high out-of-band optical density (OD). We present a comparison of a TFTF-based hyperspectral microscopy system and a commercially available AOTF-based system. We have characterized the light transmission, wavelength calibration, and OD of both systems, and have then evaluated the capability of each system for discriminating between green fluorescent protein and highly autofluorescent lung tissue. Our results suggest that TFTFs are an alternative approach for hyperspectral filtering that offers improved transmission and out-of-band blocking. These characteristics make TFTFs well suited for other biomedical imaging devices, such as ophthalmoscopes or endoscopes.

Paper Details

Date Published: 26 September 2013
PDF: 11 pages
J. Biomed. Opt. 19(1) 011017 doi: 10.1117/1.JBO.19.1.011017
Published in: Journal of Biomedical Optics Volume 19, Issue 1
Show Author Affiliations
Peter F. Favreau, Univ. of South Alabama (United States)
Clarissa Hernandez, Univ. of South Alabama (United States)
Ashley Stringfellow Lindsey, Univ. of South Alabama (United States)
Diego F. Alvarez, Univ. of South Alabama (United States)
Thomas C. Rich, Univ. of South Alabama (United States)
Prashant Prabhat, Semrock, Inc. (United States)
Silas J. Leavesley, Univ. of South Alabama (United States)

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