Integrated fluorescence and transmission electron microscopy
Integrated fluorescence and transmission electron microscopy
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DOI：
10.1016/j.jsb.2008.07.003
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发表时间：
2008-11-01
影响因子：
3
通讯作者：
Gerritsen, Hans C.
中科院分区：
文献类型：
--
作者：
Agronskaia, Alexandra V.;Valentijn, Jack A.;Gerritsen, Hans C.
相关显微镜是一种结合了荧光显微镜和电子显微镜优点的强大技术。前者可以在大视场中快速搜索感兴趣的区域，而后者在窄视场中表现出更高的分辨率。相关显微镜的常规使用受到繁琐和复杂的实验程序的严重阻碍。这部分是由于荧光显微镜和电子显微镜使用了两种不同的显微镜。本文提出了一种基于激光扫描荧光显微镜与透射电子显微镜集成的相关显微镜集成方法。使用这种方法，大大简化了对样品特征的搜索，并大大减少了进行实验的时间。在室温下，用结合小麦胚芽凝集素的AlexaFluor488标记的大鼠肠细胞，以及用抗过氧化氢酶抗体和AlexaFluor488抗体和10 nm蛋白a金免疫标记的大鼠肝脏过氧化物酶体，证明了这种综合方法的潜力。(C) 2008爱思唯尔公司版权所有。
Correlative microscopy is a powerful technique that combines the strengths of fluorescence microscopy and electron microscopy. The first enables rapid searching for regions of interest in large fields of view while the latter exhibits superior resolution over a narrow field of view. Routine use of correlative microscopy is seriously hampered by the cumbersome and elaborate experimental procedures. This is partly due to the use of two separate microscopes for fluorescence and electron microscopy. Here, an integrated approach to correlative microscopy is presented based on a laser scanning fluorescence microscope integrated in a transmission electron microscope. Using this approach the search for features in the specimen is greatly simplified and the time to carry out the experiment is strongly reduced. The potential of the integrated approach is demonstrated at room temperature on specimens of rat intestine cells labeled with AlexaFluor488 conjugated to wheat germ agglutinin and on rat liver peroxisomes immuno-labeled with anti-catalase antibodies and secondary AlexaFluor488 antibodies and 10 nm protein A-gold. (C) 2008 Elsevier Inc. All rights reserved.