In vivo imaging of unstained tissues using a compact and flexible multiphoton microendoscope
In vivo imaging of unstained tissues using a compact and flexible multiphoton microendoscope
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DOI：
10.1117/1.jbo.17.4.040505
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发表时间：
2012-04-01
影响因子：
3.5
通讯作者：
Xu, Chris
中科院分区：
文献类型：
--
作者：
Brown, Christopher M.;Rivera, David R.;Xu, Chris
我们使用紧凑且灵活的多光子显微内窥镜 (MPME) 来获取麻醉大鼠未染色的肝脏、肾脏和结肠的体内图像。该器件从光栅扫描双包层光纤 (DCF) 的纤芯发出飞秒脉冲 800 nm 光，该光通过微型梯度折射率透镜组件聚焦到组织中。来自组织的本征荧光和二次谐波生成信号通过同一 DCF 的纤芯和内包层进行外延收集。 MPME 具有外径 3 毫米、长度 4 厘米的刚性远端。图像视场尺寸为 115 μm x 115 μm，以 4.1 帧/秒、75 mW 照明功率在样品上采集。用异氟烷气体麻醉 Sprague-Dawley 大鼠，通过腹侧中线腹部切口进入组织，并用压舌板隔离器官，然后对器官进行成像。使用该设备从肝脏、肾脏和结肠采集的体内多光子图像在 75% 的成像帧中显示出与传统组织学载玻片相似的特征，没有运动伪影。据我们所知，这是使用紧凑灵活的 MPME 设备对活体未染色组织进行多光子成像的首次演示。 (C) 2012 年光电仪器工程师协会 (SPIE)。 [DOI：10 1117/1.JBO.17.4.040505]
We use a compact and flexible multiphoton microendoscope (MPME) to acquire in vivo images of unstained liver, kidney, and colon from an anesthetized rat. The device delivers femtosecond pulsed 800 nm light from the core of a raster-scanned dual-clad fiber (DCF), which is focused by a miniaturized gradient-index lens assembly into tissue. Intrinsic fluorescence and second-harmonic generation signal from the tissue is epi-collected through the core and inner clad of the same DCF. The MPME has a rigid distal tip of 3 mm in outer diameter and 4 cm in length. The image field-of-view measures 115 mu m by 115 mu m and was acquired at 4.1 frames/s with 75 mW illumination power at the sample. Organs were imaged after anesthetizing Sprague-Dawley rats with isofluorane gas, accessing tissues via a ventral-midline abdominal incision, and isolating the organs with tongue depressors. In vivo multiphoton images acquired from liver, kidney, and colon using this device show features similar to that of conventional histology slides, without motion artifact, in 75% of imaged frames. To the best of our knowledge, this is the first demonstration of multiphoton imaging of unstained tissue from a live subject using a compact and flexible MPME device. (C) 2012 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI: 10 1117/1.JBO.17.4.040505]