%0 Book Section %1 markert2017subcellular %A Markert, Sebastian M. %A Bauer, Vivien %A Muenz, Thomas S. %A Jones, Nicola G. %A Helmprobst, Frederik %A Britz, Sebastian %A Sauer, Markus %A Rössler, Wolfgang %A Engstler, Markus %A Stigloher, Christian %B Correlative Light and Electron Microscopy III %D 2017 %E Müller-Reichert, Thomas %E Verkade, Paul %I Academic Press %K ag_roessler muenz_high zoo_2 %P 21-47 %R https://doi.org/10.1016/bs.mcb.2017.03.004 %T 3D subcellular localization with superresolution array tomography on ultrathin sections of various species %U http://www.sciencedirect.com/science/article/pii/S0091679X17300481 %V 140 %X Abstract Array Tomography (AT) is a relatively easy-to-use and yet powerful method to put molecular identity in its full ultrastructural context. Ultrathin sections are stained with fluorophores and then imaged by light and afterward by electron microscopy to obtain a correlated view of a region of interest: its ultrastructure and specific staining. By combining \AT\ with high-pressure freezing for superior structural preservation and superresolution light microscopy, even small subcellular structures can be mapped in 3D. We established protocols for the application of superresolution \AT\ on ultrathin plastic sections of Caenorhabditis elegans, Trypanosoma brucei, and brain tissue of Cataglyphis fortis and Apis mellifera. All steps are described in detail from sample preparation to 3D reconstruction, including species-specific modifications. We thus showcase the versatility of our protocol and give some examples for biological questions that can be answered with this technique. We offer a step-by-step recipe for superresolution \AT\ that can be easily applied for C. elegans, T. brucei, C. fortis, and A. mellifera and adapted for other model systems.

@incollection{markert2017subcellular, abstract = {Abstract Array Tomography (AT) is a relatively easy-to-use and yet powerful method to put molecular identity in its full ultrastructural context. Ultrathin sections are stained with fluorophores and then imaged by light and afterward by electron microscopy to obtain a correlated view of a region of interest: its ultrastructure and specific staining. By combining \{AT\} with high-pressure freezing for superior structural preservation and superresolution light microscopy, even small subcellular structures can be mapped in 3D. We established protocols for the application of superresolution \{AT\} on ultrathin plastic sections of Caenorhabditis elegans, Trypanosoma brucei, and brain tissue of Cataglyphis fortis and Apis mellifera. All steps are described in detail from sample preparation to 3D reconstruction, including species-specific modifications. We thus showcase the versatility of our protocol and give some examples for biological questions that can be answered with this technique. We offer a step-by-step recipe for superresolution \{AT\} that can be easily applied for C. elegans, T. brucei, C. fortis, and A. mellifera and adapted for other model systems. }, added-at = {2017-05-03T10:58:09.000+0200}, author = {Markert, Sebastian M. and Bauer, Vivien and Muenz, Thomas S. and Jones, Nicola G. and Helmprobst, Frederik and Britz, Sebastian and Sauer, Markus and Rössler, Wolfgang and Engstler, Markus and Stigloher, Christian}, biburl = {https://www.bibsonomy.org/bibtex/2b5f69dcc8a93ffd3651af91301eb3ec4/zoologieii}, booktitle = {Correlative Light and Electron Microscopy III}, doi = {https://doi.org/10.1016/bs.mcb.2017.03.004}, editor = {Müller-Reichert, Thomas and Verkade, Paul}, interhash = {16a573944347601703937684f729c7f4}, intrahash = {b5f69dcc8a93ffd3651af91301eb3ec4}, issn = {0091-679X}, keywords = {ag_roessler muenz_high zoo_2}, pages = {21-47}, publisher = {Academic Press}, series = {Methods in Cell Biology }, timestamp = {2017-07-20T13:16:48.000+0200}, title = {3D subcellular localization with superresolution array tomography on ultrathin sections of various species }, url = {http://www.sciencedirect.com/science/article/pii/S0091679X17300481}, volume = 140, year = 2017 }