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Handheld endomicroscope using a fiber-optic harmonograph enables real-time and in vivo confocal imaging of living cell morphology and capillary perfusion
Microsystems & Nanoengineering ( IF 7.3 ) Pub Date : 2020-09-21 , DOI: 10.1038/s41378-020-00182-6
Kyungmin Hwang 1, 2 , Yeong-Hyeon Seo 1 , Daniel Y Kim 1 , Jinhyo Ahn 3 , Soyoung Lee 4 , Kyung Hee Han 5 , Koun-Hee Lee 2 , Sangyong Jon 4 , Pilhan Kim 3, 6 , Kate E Yu 2 , Hyungsin Kim 7 , Shin-Hyuk Kang 7 , Ki-Hun Jeong 1
Affiliation  

Confocal laser endomicroscopy provides high potential for noninvasive and in vivo optical biopsy at the cellular level. Here, we report a fully packaged handheld confocal endomicroscopic system for real-time, high-resolution, and in vivo cellular imaging using a Lissajous scanning fiber-optic harmonograph. The endomicroscopic system features an endomicroscopic probe with a fiber-optic harmonograph, a confocal microscope unit, and an image signal processor. The fiber-optic harmonograph contains a single mode fiber coupled with a quadrupole piezoelectric tube, which resonantly scans both axes at ~ 1 kHz to obtain a Lissajous pattern. The fiber-optic harmonograph was fully packaged into an endomicroscopic probe with an objective lens. The endomicroscopic probe was hygienically packaged for waterproofing and disinfection of medical instruments within a 2.6-mm outer diameter stainless tube capable of being inserted through the working channel of a clinical endoscope. The probe was further combined with the confocal microscope unit for indocyanine green imaging and the image signal processor for high frame rate and high density Lissajous scanning. The signal processing unit delivers driving signals for probe actuation and reconstructs confocal images using the auto phase matching process of Lissajous fiber scanners. The confocal endomicroscopic system was used to successfully obtain human in vitro fluorescent images and real-time ex vivo and in vivo fluorescent images of the living cell morphology and capillary perfusion inside a single mouse.



中文翻译:

使用光纤谐波仪的手持式内窥镜可实现活细胞形态和毛细血管灌注的实时和体内共聚焦成像

共聚焦激光内窥镜为细胞水平的无创和体内光学活检提供了很高的潜力。在这里,我们报告了一个完全封装的手持共聚焦内镜系统,用于使用 Lissajous 扫描光纤谐波进行实时、高分辨率和体内细胞成像。内窥镜系统的特点是内窥镜探头带有光纤谐波仪、共聚焦显微镜单元和图像信号处理器。光纤谐振器包含与四极压电管耦合的单模光纤,它以 ~ 1 kHz 的频率对两个轴进行共振扫描,以获得 Lissajous 模式。光纤谐波记录仪完全封装在带有物镜的内窥镜探头中。内窥镜探头采用卫生包装,用于医疗器械的防水和消毒,位于外径为 2.6 毫米的不锈钢管内,能够通过临床内窥镜的工作通道插入。该探针进一步与用于吲哚菁绿成像的共聚焦显微镜单元和用于高帧率和高密度李萨如扫描的图像信号处理器相结合。信号处理单元为探头驱动提供驱动信号,并使用 Lissajous 光纤扫描仪的自动相位匹配过程重建共焦图像。共聚焦显微系统用于成功获得人类体外荧光图像以及活细胞形态和单个小鼠毛细血管灌注的实时离体和体内荧光图像。6 毫米外径不锈钢管,能够通过临床内窥镜的工作通道插入。该探针进一步与用于吲哚菁绿成像的共聚焦显微镜单元和用于高帧率和高密度李萨如扫描的图像信号处理器相结合。信号处理单元为探头驱动提供驱动信号,并使用 Lissajous 光纤扫描仪的自动相位匹配过程重建共焦图像。共聚焦显微系统用于成功获得人类体外荧光图像以及活细胞形态和单个小鼠毛细血管灌注的实时离体和体内荧光图像。6 毫米外径不锈钢管,能够通过临床内窥镜的工作通道插入。该探针进一步与用于吲哚菁绿成像的共聚焦显微镜单元和用于高帧率和高密度李萨如扫描的图像信号处理器相结合。信号处理单元为探头驱动提供驱动信号,并使用 Lissajous 光纤扫描仪的自动相位匹配过程重建共焦图像。共聚焦显微系统用于成功获得人类体外荧光图像以及活细胞形态和单个小鼠毛细血管灌注的实时离体和体内荧光图像。该探针进一步与用于吲哚菁绿成像的共聚焦显微镜单元和用于高帧率和高密度李萨如扫描的图像信号处理器相结合。信号处理单元为探头驱动提供驱动信号,并使用 Lissajous 光纤扫描仪的自动相位匹配过程重建共焦图像。共聚焦显微系统用于成功获得人类体外荧光图像以及活细胞形态和单个小鼠毛细血管灌注的实时离体和体内荧光图像。该探针进一步与用于吲哚菁绿成像的共聚焦显微镜单元和用于高帧率和高密度李萨如扫描的图像信号处理器相结合。信号处理单元为探头驱动提供驱动信号,并使用 Lissajous 光纤扫描仪的自动相位匹配过程重建共焦图像。共聚焦显微系统用于成功获得人类体外荧光图像以及活细胞形态和单个小鼠毛细血管灌注的实时离体和体内荧光图像。

更新日期:2020-09-21
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