3-DIMENSIONAL IMAGING OF NEURONS BY CONFOCAL FLUORESCENCE MICROSCOPY
3-DIMENSIONAL IMAGING OF NEURONS BY CONFOCAL FLUORESCENCE MICROSCOPY
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DOI：
10.1111/j.1365-2818.1989.tb04296.x
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发表时间：
1989-07-01
影响因子：
2
通讯作者：
BRODIN, L
中科院分区：
文献类型：
--
作者：
CARLSSON, K;WALLEN, P;BRODIN, L
本文介绍了用激光共聚焦显微镜研究神经元结构的方法。光学连续切片已进行了整体安装标本，并产生的数字记录图像的堆栈已处理的帮助下，一台计算机。描述了样品制备，以及仪器及其性能。结果表明，测光质量的限制是由光子量子噪声。由于所研究的样品中光吸收和散射都很低，因此最大扫描深度主要受物镜工作距离的限制。与传统方法相比，共焦显微镜结合数字图像处理具有以下优点：（1）获得真正的三维（3-D）重建，（2）样品保持完整，（3）可以获得更高的分辨率，（4）过程自动化，耗时少，（5）各种数据处理是可能的。
The study of neuronal architecture by means of confocal laser microscopy is described. Optical serial sectioning has been performed on whole-mount specimens, and the resulting stacks of digitally recorded images have been processed with the help of a computer. Specimen preparation is described, as well as the instrument and its performance. It is shown that the limits in photometric quality are set by photon quantum noise. As both light absorption and scattering was low in the studied specimens, the maximum scanning depth was limited mainly by the working distance of the objectives. Compared with traditional methods, confocal microscopy in combination with digital image processing has the following advantages: (1) a truly three-dimensional (3-D) reconstruction is obtained, (2) the specimen remains intact, (3) a higher resolution can be obtained, (4) the process is automated and less time-consuming and (5) various kinds of data processing are possible.