{"id":112,"date":"2020-06-23T15:46:02","date_gmt":"2020-06-23T15:46:02","guid":{"rendered":"https:\/\/www.embl.org\/groups\/electron-microscopy-core-facility\/?page_id=112"},"modified":"2022-12-29T10:06:32","modified_gmt":"2022-12-29T10:06:32","slug":"fib-sem","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/electron-microscopy-core-facility\/services\/fib-sem\/","title":{"rendered":"Focused Ion Beam milling combined with Scanning Electron Microscopy (FIB-SEM)"},"content":{"rendered":"\n
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The EMCF has implemented a Focused Ion Beam-Scanning Electron Microscope (FIB-SEM, see figure above), together with the Schwab team and in collaboration with Carl Zeiss Microscopy. This imaging technology relies on combining serial etching of a resin block with the scanning of the exposed surface. When the cell features (membranes, cytoskeletal elements, protein complexes) are cross-sectioned, they are revealed in an almost equivalent way as with a TEM microscope. The sequential slicing and viewing steps therefore lead to the acquisition of large volumes in an automated way.<\/p>\n\n\n\n

The main advantage compared to serial TEM or serial tomography is the size of the volume that can be acquired; with a close to one thousand-fold increase in favor to the FIB-SEM.<\/p>\n\n\n\n

The fact that this technique allows imaging sub-cellular features at high resolution and in 3D opens to the study of membrane trafficking, organelle architecture and cytoskeletal arrangement in the volume of the cell, but also cell-cell or cell-matrix interactions. The goal is to make this new microscope a versatile tool for several projects. The EMCF strives to become a specialized center for the application of such approaches and will actively contribute to the methodological development necessary to make it a key technique in biology.<\/p>\n\n\n\n

3D datasets such as the one shown that was acquired acquired using the FIB-SEM from a cultured cell, allow an understanding of the volume organization within cells. Mitochondrias are segmented in red, a fraction of the ER (in blue) shows interactions with the plasma membrane specializations (green). (courtesy of Anna Steyer, Schwab team)<\/p>\n\n\n\n

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\"3D<\/a>
3D datasets such as the one shown that was acquired using the FIB-SEM from a cultured cell, allow an understanding of the volume organization within cells. Mitochondrias are segmented in red, a fraction of the ER (in blue) shows interactions with the plasma membrane specializations (green). (courtesy of Anna Steyer, Schwab team)<\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n<\/div>\n\n<\/div>\n<\/div>\n\n\n
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\"diagram\"<\/a>
In FIB-SEM microscopes, the focused ion beam is used to etch the block, exposing a surface that can be imaged by SEM (from Briggman and Bock<\/a>, Curr. Op. in Neurobiol.<\/em> 2012).<\/figcaption><\/figure>\n\n\n\n

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\"poster\"<\/a>
Automated 3DEM. Scale bars: 10\u03bcm, (from Karreman et al.<\/a> Journal of Cell Science<\/em> 2016)<\/figcaption><\/figure>\n\n\n\n

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