Institute of Chemical Sciences and Engineering (ISIC), École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, CH-1015, Switzerland.
CrestOptics S.p.A, Rome, 00167, Italy.
Sci Rep. 2018 Sep 13;8(1):13770. doi: 10.1038/s41598-018-31928-y.
Fluorescence microscopy in the second near-infrared optical window (NIR-II, 1000-1350 nm) has become a technique of choice for non-invasive in vivo imaging. The deep penetration of NIR light in living tissue, as well as negligible tissue autofluorescence within this optical range, offers increased resolution and contrast with even greater penetration depths. Here, we present a custom-built spinning-disc confocal laser microscope (SDCLM) that is specific to imaging in the NIR-II. The SDCLM achieves a lateral resolution of 0.5 ± 0.1 µm and an axial resolution of 0.6 ± 0.1 µm, showing a ~17% and ~45% enhancement in lateral and axial resolution, respectively, compared to the corresponding wide-field configuration. We furthermore showcase several applications that demonstrate the use of the SDCLM for in situ, spatiotemporal tracking of NIR particles and bioanalytes within both synthetic and biological systems.
荧光显微镜在近红外二区（NIR-II，1000-1350nm）已经成为一种非侵入式活体成像的首选技术。近红外光在活体组织中的深穿透性，以及在此光学范围内可忽略的组织自发荧光，提供了更高的分辨率和对比度，甚至具有更大的穿透深度。在这里，我们展示了一种专门针对 NIR-II 成像的定制旋转盘共聚焦激光显微镜（SDCLM）。SDCLM 实现了 0.5±0.1μm 的横向分辨率和 0.6±0.1μm 的轴向分辨率，与相应的宽场配置相比，分别提高了约 17%和 45%的横向和轴向分辨率。我们还展示了几个应用，展示了 SDCLM 在原位、时空追踪合成和生物体系中 NIR 粒子和生物分析物的用途。
