Thin-film tunable filters for hyperspectral fluorescence microscopy
Thin-film tunable filters for hyperspectral fluorescence microscopy
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DOI：
10.1117/1.jbo.19.1.011017
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发表时间：
2014-01-01
影响因子：
3.5
通讯作者：
Leavesley, Silas J.
中科院分区：
文献类型：
--
作者：
Favreau, Peter;Hernandez, Clarissa;Leavesley, Silas J.
高光谱成像是一种强大的工具，可以从许多光谱带获取数据，形成连续的光谱。高光谱成像最初是为遥感应用而开发的；然而，高光谱技术已应用于生物荧光成像应用，例如荧光显微镜和小动物荧光成像。光谱滤波方法在很大程度上决定了任何高光谱成像系统的灵敏度和特异性。有多种类型的光谱滤波硬件可用于显微镜系统，最常见的是声光可调谐滤波器 (AOTF) 和液晶可调谐滤波器 (LCTF)。这些过滤技术各有优点和缺点。在这里，我们提出了一种用于高光谱成像的新型可调谐滤波器——薄膜可调谐滤波器（TFTF）。与 AOTF 和 LCTF 相比，TFTF 具有多种优势，最显着的是高百分比透射率和高带外光密度 (OD)。我们对基于 TFTF 的高光谱显微镜系统和市售的基于 AOTF 的系统进行了比较。我们对两个系统的光传输、波长校准和 OD 进行了表征，然后评估了每个系统区分绿色荧光蛋白和高度自发荧光肺组织的能力。我们的结果表明，TFTF 是高光谱滤波的一种替代方法，可提供改进的传输和带外阻塞。这些特性使 TFTF 非常适合其他生物医学成像设备，例如检眼镜或内窥镜。 (C) 2014 年光电仪器工程师学会 (SPIE)
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. (C) 2014 Society of Photo-Optical Instrumentation Engineers (SPIE)