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Biomedical Imaging: Principles and Applications by Reiner Salzer

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Chapter 6

Toward A 3D View of Cellular Architecture: Correlative Light Microscopy and Electron Tomography

Jack A. Valentijn, Linda F. van Driel, Karen A. Jansen, Karine M. Valentijn, and Abraham J. Koster

Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands

6.1 Introduction

The terms “multimodality imaging” and “correlative microscopy” are employed in the biomedical literature to designate any combination of two or more microscopic techniques applied to the same region in a biological specimen. “Correlative microscopy” should not be confused with “fluorescence correlation microscopy,” which is a method to measure diffusion of fluorescent molecules in cells (Brock and Jovin, 1998). The purpose of multimodality imaging is to obtain complementary data, each imaging modality providing different information on the specimen that is under investigation. Correlative Light and Electron Microscopy (CLEM) is by far the most widespread form of multimodality imaging.

CLEM makes use of the fact that imaging with photons on one hand, and electrons on the other, offers specific advantages over one another. For instance, the low magnification range inherent to Light Microscopy (LM) is particularly well suited for the rapid scanning of large and heterogeneous sample areas, while the high resolution that can be achieved by Electron Microscopy (EM) allows for the subsequent zooming in on selected areas of interest to obtain ultrastructural detail. A further ...

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