Automated imaging of extended tissue volumes using confocal microscopy
Automated imaging of extended tissue volumes using confocal microscopy
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DOI：
10.1002/jemt.20200
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发表时间：
2005-08-01
影响因子：
2.5
通讯作者：
Legrice, IJ
中科院分区：
文献类型：
--
作者：
Sands, GB;Gerneke, DA;Legrice, IJ
共聚焦显微镜能够以高分辨率观察细胞和组织的组成元素，并在三维空间中重建，但受限于可以成像的体积的有限范围。我们已经开发了一种自动化技术，可以在大的组织区域和体积上获得连续共聚焦图像。计算机控制的系统集成了共聚焦显微镜和使用高精度平移台的超磨机，固有地保留了标本的配准，并且用户控制界面可以在广泛的尺度和分辨率范围内灵活地规范成像协议。有了这个系统，就可以在三维重建指定的形态特征，并在整个组织样本中准确地定位它们。我们已经成功地在4毫米(3)的体积和75毫米的面积上以1 μ m体素分辨率对各种样品进行了成像。与适当的包埋介质和免疫组织化学探针结合使用，本文中描述的技术使得在更大的组织域上常规绘制关键细胞内结构的分布成为可能，这在过去是很容易实现的。
Confocal microscopy enables constitutive elements of cells and tissues to be viewed at high resolution and reconstructed in three dimensions, but is constrained by the limited extent of the volumes that can be imaged. We have developed an automated technique that enables serial confocal images to be acquired over large tissue areas and volumes. The computer-controlled system, which integrates a confocal microscope and an ultramill using a high-precision translation stage, inherently preserves specimen registration, and the user control interface enables flexible specification of imaging protocols over a wide range of scales and resolutions. With this system it is possible to reconstruct specified morphological features in three dimensions and locate them accurately throughout a tissue sample. We have successfully imaged various samples at 1-mu m voxel resolution on volumes up to 4 mm(3) and on areas up to 75 mm. Used in conjunction with appropriate embedding media and immuno-histochemical probes, the techniques described in this paper make it possible to routinely map the distributions of key intracellular structures over much larger tissue domains than has been easily achievable in the past.