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Journal of Electron Microscopy 51:113-126 (2002)
© 2002 Oxford University Press

Computer-assisted simulation of high-voltage electron microscopy using serial images recorded by conventional transmission electron microscopy

Ragnar Pascher 1,* , Claes-Henric Berthold2 and Martin Rydmark1,2

1MedNet, The Computer Laboratory of the Faculty of Medicine and 2Section of Neuroanatomy, Department of Anatomy and Cell Biology, Göteborg University, Box 417, SE-405 30 Göteborg, Sweden

*To whom correspondence should be addressed. E-mail: ragnar.pascher{at}mednet.gu.se

Here we describe a computer-assisted method which, based on conventional transmission electron microscopy, renders simulated high-voltage electron micrographs. We perform arithmetic minimum filtering on stacks of aligned serial transmission electron microscopic images. In this way, the structural information of the separate images is fused into one compound image that highlights organization patterns otherwise easily overlooked or impossible to comprehend. Our method, like high-voltage electron microscopy, offers the possibility to build stereo-pairs for high-resolution three-dimensional analysis of tissue layers 1–2 µm thick. The use of background elimination and the development of a depth enhancement routine improved the three-dimensional effect and facilitated the analysis of the interior of objects. As an example, we use the method to display the distribution of axonal organelles at nodes of Ranvier and the shape and contents of a highly branched hippocampal dendritic spine.

Keywords     electron microscopy, image analysis, stereographic, node of Ranvier, dendritic spine


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