In vivo label-free confocal imaging of the deep mouse brain with long-wavelength illumination.
In vivo label-free confocal imaging of the deep mouse brain with long-wavelength illumination.
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DOI：
10.1364/boe.9.006545
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发表时间：
2018-11
影响因子：
3.4
通讯作者：
Fei Xia;Chunyan Wu;D. Sinefeld;Bo Li;Yifan Qin;Chris Xu
中科院分区：
文献类型：
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作者：
Fei Xia;Chunyan Wu;D. Sinefeld;Bo Li;Yifan Qin;Chris Xu
光学显微镜是用于体内监测生物结构和功能的有价值工具，因为其无侵入性。然而，由于光的散射和吸收，深入生物组织的成像是具有挑战性的。先前的研究表明，1300 nm和1700 nm是深脑成像的两个最佳波长窗口。在这里，我们将〜1700 nm的长波长照明与反射率共聚焦显微镜相结合，并在成年小鼠大脑中获得了〜1.3 mm的成像深度，并在成年小鼠大脑中的空间分辨率约为1.3 mm，该空间分辨率比使用可见波长的常规共焦显微镜深3-4倍。我们表明，可以将该方法添加到任何具有简单和低成本源和检测器的激光扫描显微镜中，例如连续波二极管激光器和Ingaas光电二极管。我们证明的长波长，反射率共焦成像不含标签，需要低照明能力。此外，成像系统是简单且低成本的，有可能为生物医学研究和临床应用创造新的机会。
Optical microscopy is a valuable tool for in vivo monitoring of biological structures and functions because of its non-invasiveness. However, imaging deep into biological tissues is challenging due to the scattering and absorption of light. Previous research has shown that 1300 nm and 1700 nm are the two best wavelength windows for deep brain imaging. Here, we combined long-wavelength illumination of ~1700 nm with reflectance confocal microscopy and achieved an imaging depth of ~1.3 mm with ~1-micrometer spatial resolution in adult mouse brains, which is 3-4 times deeper than that of conventional confocal microscopy using visible wavelength. We showed that the method can be added to any laser-scanning microscopy with simple and low-cost sources and detectors, such as continuous-wave diode lasers and InGaAs photodiodes. The long-wavelength, reflectance confocal imaging we demonstrated is label-free, and requires low illumination power. Furthermore, the imaging system is simple and low-cost, potentially creating new opportunities for biomedical research and clinical applications.