Development of a versatile two-photon endoscope for biological imaging.
Development of a versatile two-photon endoscope for biological imaging.
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DOI：
10.1364/boe.1.001159
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发表时间：
2010-10-13
影响因子：
3.4
通讯作者：
Gordon RJ
中科院分区：
文献类型：
--
作者：
Zhao Y;Nakamura H;Gordon RJ
我们描述了一种多功能的，导管型双光子探针，设计用于在体内和体外成像的水流出途径的眼睛。该装置由用于激光传输和荧光收集的石英双包层光纤、由微型压电管驱动的螺旋光纤扫描器和用于聚焦激光和收集双光子激发信号的三个微尺寸双合消色差透镜组成。所有组件的最大直径为2 mm，并封装在外径为2.8 mm的12号不锈钢皮下注射管中。测得探头的横向和轴向分辨率分别为1.5 μm和9.2 μm。不同的透镜配置和光纤进行评估，通过比较它们的空间分辨率和荧光信号收集效率。双消色差透镜和具有高内包层数值孔径的双包层光纤被发现产生高的信号收集效率，这对于活体组织成像是必要的。减少图像失真的简单方法进行了演示。利用该微型双光子显微镜成功地获得了人眼小梁组织的图像。
We describe a versatile, catheter-type two-photon probe, designed for in vivo and ex vivo imaging of the aqueous outflow pathway in the eye. The device consists of a silica double cladding fiber used for laser delivery and fluorescence collection, a spiral fiber scanner driven by a miniature piezoelectric tube, and an assembly of three micro-size doublet achromatic lenses used for focusing the laser and collecting the two-photon excitation signal. All the components have a maximum diameter of 2 mm and are enclosed in a length of 12-gauge stainless steel hypodermic tubing having an outer diameter of 2.8 mm. The lateral and axial resolutions of the probe are measured to be 1.5 μm and 9.2 μm, respectively. Different lens configurations and fibers are evaluated by comparing their spatial resolutions and fluorescence signal collection efficiencies. Doublet achromatic lenses and a double cladding fiber with a high inner cladding numerical aperture are found to produce a high signal collection efficiency, which is essential for imaging live tissues. Simple methods for reducing image distortions are demonstrated. Images of human trabecular meshwork tissue are successfully obtained with this miniature two-photon microscope.