Correlative Light and Volume Electron Microscopy: Methods and Applications

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Background

Correlative light and electron microscopy was developed in the 1960s to bridge the resolution gap between the two imaging technologies. The purpose of such studies was to link the localization and functional studies that could be easily done at light microscopic resolution to cellular ultrastructure. In the last two decades such studies have been extended from two to three dimensions with the development of volume electron microscopy technologies, allowing the morphological and functional information to be visualized in the context of the complex nano-environment of cells and tissues. Correlative light and volume electron microscopy (vCLEM) now includes multi-modal imaging techniques such as elemental analysis, micro mass spectroscopy, X-Ray imaging and molecular structure cryo-tomography and links structures and function in ways heretofore impossible.

Cells and tissues are three dimensional structures and fully understanding their function in health and disease requires that the complex interactions between proteins and cellular components be visualized within the context of their micro-environment. Advances in 3D imaging in both the light and electron microscope have made correlative light and volume electron microscopy (vCLEM) a practical and efficient technique. When cells are visualized in 3D many assumptions made using 2D data have been modified or even rejected outright. vCLEM studies are frequently complex and multi-modal, linking many imaging and analysis techniques. Breakthroughs in vCLEM imaging techniques alongside cutting edge image reconstruction and analysis can provide a more complete picture of how cells work, and how they react to disease, trauma or infection.

Correlative light and volume electron microscopy (vCLEM) represents a multi-scale, multi-modal approach to imaging cells in ways that link function to structure. Submissions can be original research studies on a specific scientific question using vCLEM, new morphological insights obtained from 3D visualization, a new workflow type (e.g. super-resolution LM, X-Ray and FIB-SEM), a technological advance in vCLEM (e.g. cryo fluorescence LM to Cryo FIB-SEM), new methods for image analysis and visualization or a review of a specific multi-modal technique such as correlative LM and volume EDS.

Keywords: Correlative light and electron microscopy, Volume electron microscopy, Serial Block Face Scanning Electron Microscopy, Focused Ion Beam Scanning Electron, Microscopy, Array Tomography, Image Analysis

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