Thin-film tunable filters for hyperspectral fluorescence microscopy

Peter F. Favreau; Clarissa Hernandez; Ashley Stringfellow Lindsey; Diego F. Alvarez; Thomas C. Rich; Prashant Prabhat; Silas J. Leavesley

doi:10.1117/1.JBO.19.1.011017

26 September 2013 Thin-film tunable filters for hyperspectral fluorescence microscopy

Peter F. Favreau, Clarissa Hernandez, Ashley Stringfellow Lindsey, Diego F. Alvarez, Thomas C. Rich, Prashant Prabhat, Silas J. Leavesley

Author Affiliations +

Journal of Biomedical Optics, Vol. 19, Issue 1, 011017 (September 2013). https://doi.org/10.1117/1.JBO.19.1.011017

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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Peter F. Favreau, Clarissa Hernandez, Ashley Stringfellow Lindsey, Diego F. Alvarez, Thomas C. Rich, Prashant Prabhat, and Silas J. Leavesley "Thin-film tunable filters for hyperspectral fluorescence microscopy," Journal of Biomedical Optics 19(1), 011017 (26 September 2013). https://doi.org/10.1117/1.JBO.19.1.011017

Published: 26 September 2013

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KEYWORDS

Tunable filters

Luminescence

Optical filters

Green fluorescent protein

Microscopy

Thin films

Hyperspectral imaging

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