Special Section on Advanced Biomedical Imaging and Sensing

Thin-film tunable filters for hyperspectral fluorescence microscopy

[+] Author Affiliations
Peter Favreau

University of South Alabama, Department of Chemical and Biomolecular Engineering, Mobile, Alabama 36688

University of South Alabama, Center for Lung Biology, Mobile, Alabama 36688

Clarissa Hernandez

University of South Alabama, Department of Chemical and Biomolecular Engineering, Mobile, Alabama 36688

Ashley Stringfellow Lindsey

University of South Alabama, Department of Pharmacology, Mobile, Alabama 36688

Diego F. Alvarez

University of South Alabama, Department of Pharmacology, Mobile, Alabama 36688

University of South Alabama, Department of Internal Medicine, Mobile, Alabama 36688

University of South Alabama, Center for Lung Biology, Mobile, Alabama 36688

Thomas Rich

University of South Alabama, Department of Pharmacology, Mobile, Alabama 36688

University of South Alabama, Center for Lung Biology, Mobile, Alabama 36688

Prashant Prabhat

Semrock, Inc. (A Unit of IDEX Corporation), 3625 Buffalo Road, Rochester, New York 14624

Silas J. Leavesley

University of South Alabama, Department of Chemical and Biomolecular Engineering, Mobile, Alabama 36688

University of South Alabama, Department of Pharmacology, Mobile, Alabama 36688

University of South Alabama, Center for Lung Biology, Mobile, Alabama 36688

J. Biomed. Opt. 19(1), 011017 (Sep 26, 2013). doi:10.1117/1.JBO.19.1.011017
History: Received March 7, 2013; Revised August 27, 2013; Accepted August 28, 2013
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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.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Peter Favreau ; Clarissa Hernandez ; Ashley Stringfellow Lindsey ; Diego F. Alvarez ; Thomas Rich, et al.
"Thin-film tunable filters for hyperspectral fluorescence microscopy", J. Biomed. Opt. 19(1), 011017 (Sep 26, 2013). ; http://dx.doi.org/10.1117/1.JBO.19.1.011017


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