Chazot Cécile A C, Nagelberg Sara, Rowlands Christopher J, Scherer Maik R J, Coropceanu Igor, Broderick Kurt, Kim Yunjo, Bawendi Moungi G, So Peter, Kolle Mathias
Mechanical Engineering Department, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts, 02139, United States of America.
Department of Bioengineering, Imperial College London, South Kensington, SW7 2AZ, UK.
Nat Photonics. 2020 May;14(5):310-315. doi: 10.1038/s41566-020-0593-1. Epub 2020 Feb 24.
Dark-field microscopy is a standard imaging technique widely employed in biology that provides high image contrast for a broad range of unstained specimens. Unlike bright-field microscopy, it accentuates high spatial frequencies and can therefore be used to emphasize and resolve small features. However, the use of dark-field microscopy for reliable analysis of blood cells, bacteria, algae, and other marine organisms often requires specialized, bulky microscope systems, and expensive additional components, such as dark-field-compatible objectives or condensers. Here, we propose to simplify and downsize dark-field microscopy equipment by generating the high-angle illumination cone required for dark field microscopy directly within the sample substrate. We introduce a luminescent photonic substrate with a controlled angular emission profile and demonstrate its ability to generate high-contrast dark-field images of micrometre-sized living organisms using standard optical microscopy equipment. This new type of substrate forms the basis for miniaturized lab-on-chip dark-field imaging devices, compatible with simple and compact light microscopes.
暗场显微镜是生物学中广泛使用的一种标准成像技术,可为各种未染色的标本提供高图像对比度。与明场显微镜不同,它突出高空间频率,因此可用于强调和分辨小特征。然而,将暗场显微镜用于血细胞、细菌、藻类和其他海洋生物的可靠分析通常需要专门的、体积庞大的显微镜系统以及昂贵的附加组件,如与暗场兼容的物镜或聚光镜。在此,我们提议通过直接在样品基板内生成暗场显微镜所需的高角度照明锥来简化和缩小暗场显微镜设备的尺寸。我们引入了一种具有可控角发射轮廓的发光光子基板,并展示了其使用标准光学显微镜设备生成微米级活生物体高对比度暗场图像的能力。这种新型基板构成了与简单紧凑的光学显微镜兼容的微型芯片实验室暗场成像设备的基础。