Automated microscopy system for mosaic acquisition and processing
Automated microscopy system for mosaic acquisition and processing
复制标题
DOI：
10.1111/j.1365-2818.2006.01577.x
复制
发表时间：
2006-05-01
影响因子：
2
通讯作者：
Ellisman, M. H.
中科院分区：
文献类型：
--
作者：
Chow, S. K.;Hakozaki, H.;Ellisman, M. H.
描述了一种用于多光子显微镜的自动镶嵌采集和处理系统，用于在光学显微镜的分辨率极限处或接近光学显微镜的分辨率极限处对大面积的生物样品进行成像。在马赛克中，较大的图像是从一系列较小的图像中创建的，这些图像是在样本上系统地单独采集的。马赛克允许在不牺牲分辨率的情况下获得生物标本的宽视场视图，提供背景下生物标本的详细视图。该系统是由一个快速扫描，多光子，共聚焦显微镜配有一个机动，高精度的阶段和定制开发的软件程序，自动图像采集，图像归一化，图像对齐和拼接。我们目前的能力使我们能够获得由每个马赛克数千到数万个单独图像组成的数据集。创建单个马赛克所涉及的大量单个图像需要软件开发来自动化马赛克采集和处理步骤。在这份报告中，我们描述的方法和挑战，涉及常规创建非常大规模的马赛克从脑组织标记有多种荧光探针。
An automatic mosaic acquisition and processing system for a multiphoton microscope is described for imaging large expanses of biological specimens at or near the resolution limit of light microscopy. In a mosaic, a larger image is created from a series of smaller images individually acquired systematically across a specimen. Mosaics allow wide-field views of biological specimens to be acquired without sacrificing resolution, providing detailed views of biological specimens within context. The system is composed of a fast-scanning, multiphoton, confocal microscope fitted with a motorized, high-precision stage and custom-developed software programs for automatic image acquisition, image normalization, image alignment and stitching. Our current capabilities allow us to acquire data sets comprised of thousands to tens of thousands of individual images per mosaic. The large number of individual images involved in creating a single mosaic necessitated software development to automate both the mosaic acquisition and processing steps. In this report, we describe the methods and challenges involved in the routine creation of very large scale mosaics from brain tissue labelled with multiple fluorescent probes.