Miniaturized selective plane illumination microscopy for high-contrast in vivo fluorescence imaging
Miniaturized selective plane illumination microscopy for high-contrast in vivo fluorescence imaging
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DOI：
10.1364/ol.35.001413
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发表时间：
2010-05-01
期刊：
影响因子：
3.6
通讯作者：
Helmchen, Fritjof
中科院分区：
文献类型：
--
作者：
Engelbrecht, Christoph J.;Voigt, Fabian;Helmchen, Fritjof
基于光片的荧光成像技术依赖于单个光学平面的同时激发，从而允许对扩展的组织样本进行高对比度光学切片成像。在这里，我们介绍了一种选择平面照明显微镜(Mini SPIM)的小型化光纤实现。激发光通过单模光纤传输，并用柱面梯度折射率透镜和直角微棱镜产生光片。荧光发射通过梯度折射率透镜组件和相干光纤束与光片垂直收集。纤维束的端面被成像到电荷耦合设备相机上。微型SPIM的空间分辨率横向为3.2微米，轴向为5.1微米。在微型SPIM模式下，小鼠新皮质中荧光珠和神经元的图像比在表观照明模式下记录的图像显示出更高的轴向分辨率和对比度。因此，微型SPIM可能会使新的体内成像方法成为可能。(C)2010年美国光学学会
Light-sheet-based fluorescence imaging techniques rely on simultaneous excitation of a single optical plane and thus permit high-contrast optically sectioned imaging of extended tissue samples. Here, we introduce a miniaturized fiber-optic implementation of a selective plane-illumination microscope (miniSPIM). The excitation light was delivered through a single-mode optical fiber, and a light-sheet was created with a cylindrical gradient-index lens and a right-angle microprism. Fluorescence emission was collected orthogonally to the light-sheet through a gradient-index lens assembly and a coherent fiber bundle. The end face of the fiber bundle was imaged onto a charge-coupled device camera. The spatial resolutions of the miniSPIM were 3.2 mu m laterally and 5.1 mu m axially. Images of fluorescent beads and neurons in mouse neocortex exhibited superior axial resolution and contrast in the miniSPIM-mode compared to images recorded in epi-illumination mode. The miniSPIM may thus enable novel in vivo imaging approaches. (C) 2010 Optical Society of America