Atmospheric Scanning Electron Microscopy (ASEM) Realizes Direct EM-OM Linkage in Solution: Aqueous Immuno-Cytochemistry

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JEOLnews Volume 46, Number 1,2011 Chikara Sato

Biomedical Research Institute, National Institute of
Advanced Industrial Science and Technology (AIST)

The JEOL ClairScope™ Atmospheric SEM (ASEM) is the next generation in environmental EM, inverting the traditional SEM column to view the sample from beneath. The open ASEM dish on the top of the column allows directly linked correlative microscopy in open solution: optical microscope (OM) images from above, immediately followed by EM of the same sample area at high resolution, from below. The 35 mm ASEM dish allows neuronal primary culture and efficient antibody labeling-washing cycles. Because the epitopes of cells are preserved in aqueous solution, we tested antibodies, including mouse monoclonal antibodies, used in immuno-fluorescence microscopy. All were successful, enabling observation at SEM resolutions. The resolution of the ASEM is 8 nm, which is advantageous for the observation of fine intracellular structures and bacteria. Observable depth is 2-3 μm at 30 kV, and tissue sections placed on the ASEM dish can be observed immediately. Stained with platinum blue, cell nuclei were especially prominent. This is applicable not only to basic biology, but also to intra-operative cancer diagnosis and detection of pathogens.The ASEM is also highly applicable in non-bioscience fields including materials sciences.
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