Sasagawa Kiyotaka, Kimura Ayaka, Haruta Makito, Noda Toshihiko, Tokuda Takashi, Ohta Jun
Division of Materials Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, Japan.
Biomed Opt Express. 2018 Aug 17;9(9):4329-4344. doi: 10.1364/BOE.9.004329. eCollection 2018 Sep 1.
We report a lens-free fluorescence imaging device using a composite filter composed of an interference filter and an absorption filter, each applied to one side of a fiber optic plate (FOP). The transmission of angled excitation light through the interference filter is absorbed by the absorption filter. The auto-fluorescence of the absorption filter is reduced by the reflection from the interference filter of normally incident excitation light. As a result, high-performance rejection of excitation light is achieved in a lens-free device. The FOP provides a flat, hard imaging device surface that does not degrade the spatial resolution. We demonstrate excitation rejection of approximately 10:1 at a wavelength of 450 nm in a fabricated lens-free device. The resolution of fluorescence imaging is approximately 12 m. Time-lapse imaging of cells containing green fluorescent protein was performed in a 5-m thin-film chamber. The small dimensions of the device allow observation of cell culturing in a CO incubator. We also demonstrate that the proposed lens-free filter is compatible with super-resolution bright-field imaging techniques. These features open a way to develop a high-performance, dual-mode, lens-free imaging device that is expected to be a powerful tool for many applications, such as imaging of labeled cells and point-of-care assay.
我们报道了一种无透镜荧光成像设备,该设备使用了一种由干涉滤光片和吸收滤光片组成的复合滤光片,每个滤光片分别应用于光纤板(FOP)的一侧。倾斜的激发光透过干涉滤光片后的透射光被吸收滤光片吸收。吸收滤光片的自发荧光通过正常入射激发光在干涉滤光片上的反射而降低。结果,在无透镜设备中实现了对激发光的高性能抑制。光纤板提供了一个平坦、坚硬的成像设备表面,不会降低空间分辨率。我们在制造的无透镜设备中展示了在450 nm波长下约10:1的激发光抑制率。荧光成像的分辨率约为12μm。在一个5μm的薄膜腔室中对含有绿色荧光蛋白的细胞进行了延时成像。该设备的小尺寸允许在CO₂培养箱中观察细胞培养。我们还证明了所提出的无透镜滤光片与超分辨率明场成像技术兼容。这些特性为开发一种高性能、双模式、无透镜成像设备开辟了道路,该设备有望成为许多应用的强大工具,如标记细胞成像和即时检测。