[{"publisher":"Elsevier","page":"33-56","volume":161,"date_published":"2021-01-01T00:00:00Z","external_id":{"pmid":["33478696"]},"status":"public","date_created":"2020-06-07T22:00:55Z","month":"01","publication_identifier":{"issn":["0091-679X"],"isbn":["978012820807-6"]},"scopus_import":"1","intvolume":"       161","day":"01","year":"2021","date_updated":"2021-03-11T08:49:08Z","type":"book_chapter","oa_version":"None","article_processing_charge":"No","author":[{"full_name":"Truckenbrodt, Sven M","id":"45812BD4-F248-11E8-B48F-1D18A9856A87","last_name":"Truckenbrodt","first_name":"Sven M"},{"full_name":"Rizzoli, Silvio O.","first_name":"Silvio O.","last_name":"Rizzoli"}],"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","quality_controlled":"1","citation":{"apa":"Truckenbrodt, S. M., &#38; Rizzoli, S. O. (2021). Simple multi-color super-resolution by X10 microscopy. In <i>Methods in Cell Biology</i> (Vol. 161, pp. 33–56). Elsevier. <a href=\"https://doi.org/10.1016/bs.mcb.2020.04.016\">https://doi.org/10.1016/bs.mcb.2020.04.016</a>","mla":"Truckenbrodt, Sven M., and Silvio O. Rizzoli. “Simple Multi-Color Super-Resolution by X10 Microscopy.” <i>Methods in Cell Biology</i>, vol. 161, Elsevier, 2021, pp. 33–56, doi:<a href=\"https://doi.org/10.1016/bs.mcb.2020.04.016\">10.1016/bs.mcb.2020.04.016</a>.","short":"S.M. Truckenbrodt, S.O. Rizzoli, in:, Methods in Cell Biology, Elsevier, 2021, pp. 33–56.","chicago":"Truckenbrodt, Sven M, and Silvio O. Rizzoli. “Simple Multi-Color Super-Resolution by X10 Microscopy.” In <i>Methods in Cell Biology</i>, 161:33–56. Elsevier, 2021. <a href=\"https://doi.org/10.1016/bs.mcb.2020.04.016\">https://doi.org/10.1016/bs.mcb.2020.04.016</a>.","ista":"Truckenbrodt SM, Rizzoli SO. 2021.Simple multi-color super-resolution by X10 microscopy. In: Methods in Cell Biology. vol. 161, 33–56.","ama":"Truckenbrodt SM, Rizzoli SO. Simple multi-color super-resolution by X10 microscopy. In: <i>Methods in Cell Biology</i>. Vol 161. Elsevier; 2021:33-56. doi:<a href=\"https://doi.org/10.1016/bs.mcb.2020.04.016\">10.1016/bs.mcb.2020.04.016</a>","ieee":"S. M. Truckenbrodt and S. O. Rizzoli, “Simple multi-color super-resolution by X10 microscopy,” in <i>Methods in Cell Biology</i>, vol. 161, Elsevier, 2021, pp. 33–56."},"publication_status":"published","abstract":[{"text":"Expansion microscopy is a recently developed super-resolution imaging technique, which provides an alternative to optics-based methods such as deterministic approaches (e.g. STED) or stochastic approaches (e.g. PALM/STORM). The idea behind expansion microscopy is to embed the biological sample in a swellable gel, and then to expand it isotropically, thereby increasing the distance between the fluorophores. This approach breaks the diffraction barrier by simply separating the emission point-spread-functions of the fluorophores. The resolution attainable in expansion microscopy is thus directly dependent on the separation that can be achieved, i.e. on the expansion factor. The original implementation of the technique achieved an expansion factor of fourfold, for a resolution of 70–80 nm. The subsequently developed X10 method achieves an expansion factor of 10-fold, for a resolution of 25–30 nm. This technique can be implemented with minimal technical requirements on any standard fluorescence microscope, and is more easily applied for multi-color imaging than either deterministic or stochastic super-resolution approaches. This renders X10 expansion microscopy a highly promising tool for new biological discoveries, as discussed here, and as demonstrated by several recent applications.","lang":"eng"}],"doi":"10.1016/bs.mcb.2020.04.016","language":[{"iso":"eng"}],"department":[{"_id":"JoDa"}],"_id":"7941","title":"Simple multi-color super-resolution by X10 microscopy","publication":"Methods in Cell Biology","pmid":1},{"scopus_import":"1","date_published":"2019-03-01T00:00:00Z","external_id":{"pmid":["30778205"],"isi":["000459890700008"]},"file":[{"success":1,"file_id":"9619","date_updated":"2021-06-29T14:41:46Z","file_name":"181031_Truckenbrodt_ExM_NatProtoc.docx","checksum":"7efb9951e7ddf3e3dcc2fb92b859c623","access_level":"open_access","relation":"main_file","date_created":"2021-06-29T14:41:46Z","file_size":84478958,"creator":"kschuh","content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document"}],"page":"832–863","ec_funded":1,"doi":"10.1038/s41596-018-0117-3","language":[{"iso":"eng"}],"has_accepted_license":"1","department":[{"_id":"JoDa"},{"_id":"Bio"}],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","article_processing_charge":"No","date_updated":"2023-08-24T14:48:33Z","type":"journal_article","oa_version":"Submitted Version","day":"01","intvolume":"        14","isi":1,"date_created":"2019-02-24T22:59:20Z","month":"03","status":"public","volume":14,"issue":"3","file_date_updated":"2021-06-29T14:41:46Z","publisher":"Nature Publishing Group","article_type":"original","title":"A practical guide to optimization in X10 expansion microscopy","publication":"Nature Protocols","pmid":1,"oa":1,"ddc":["570"],"_id":"6052","project":[{"grant_number":"754411","name":"ISTplus - Postdoctoral Fellowships","call_identifier":"H2020","_id":"260C2330-B435-11E9-9278-68D0E5697425"},{"call_identifier":"FWF","_id":"265CB4D0-B435-11E9-9278-68D0E5697425","grant_number":"I03600","name":"Optical control of synaptic function via adhesion molecules"}],"publication_status":"published","abstract":[{"text":"Expansion microscopy is a relatively new approach to super-resolution imaging that uses expandable hydrogels to isotropically increase the physical distance between fluorophores in biological samples such as cell cultures or tissue slices. The classic gel recipe results in an expansion factor of ~4×, with a resolution of 60–80 nm. We have recently developed X10 microscopy, which uses a gel that achieves an expansion factor of ~10×, with a resolution of ~25 nm. Here, we provide a step-by-step protocol for X10 expansion microscopy. A typical experiment consists of seven sequential stages: (i) immunostaining, (ii) anchoring, (iii) polymerization, (iv) homogenization, (v) expansion, (vi) imaging, and (vii) validation. The protocol presented here includes recommendations for optimization, pitfalls and their solutions, and detailed guidelines that should increase reproducibility. Although our protocol focuses on X10 expansion microscopy, we detail which of these suggestions are also applicable to classic fourfold expansion microscopy. We exemplify our protocol using primary hippocampal neurons from rats, but our approach can be used with other primary cells or cultured cell lines of interest. This protocol will enable any researcher with basic experience in immunostainings and access to an epifluorescence microscope to perform super-resolution microscopy with X10. The procedure takes 3 d and requires ~5 h of actively handling the sample for labeling and expansion, and another ~3 h for imaging and analysis.","lang":"eng"}],"citation":{"ieee":"S. M. Truckenbrodt, C. M. Sommer, S. O. Rizzoli, and J. G. Danzl, “A practical guide to optimization in X10 expansion microscopy,” <i>Nature Protocols</i>, vol. 14, no. 3. Nature Publishing Group, pp. 832–863, 2019.","ama":"Truckenbrodt SM, Sommer CM, Rizzoli SO, Danzl JG. A practical guide to optimization in X10 expansion microscopy. <i>Nature Protocols</i>. 2019;14(3):832–863. doi:<a href=\"https://doi.org/10.1038/s41596-018-0117-3\">10.1038/s41596-018-0117-3</a>","chicago":"Truckenbrodt, Sven M, Christoph M Sommer, Silvio O Rizzoli, and Johann G Danzl. “A Practical Guide to Optimization in X10 Expansion Microscopy.” <i>Nature Protocols</i>. Nature Publishing Group, 2019. <a href=\"https://doi.org/10.1038/s41596-018-0117-3\">https://doi.org/10.1038/s41596-018-0117-3</a>.","short":"S.M. Truckenbrodt, C.M. Sommer, S.O. Rizzoli, J.G. Danzl, Nature Protocols 14 (2019) 832–863.","ista":"Truckenbrodt SM, Sommer CM, Rizzoli SO, Danzl JG. 2019. A practical guide to optimization in X10 expansion microscopy. Nature Protocols. 14(3), 832–863.","mla":"Truckenbrodt, Sven M., et al. “A Practical Guide to Optimization in X10 Expansion Microscopy.” <i>Nature Protocols</i>, vol. 14, no. 3, Nature Publishing Group, 2019, pp. 832–863, doi:<a href=\"https://doi.org/10.1038/s41596-018-0117-3\">10.1038/s41596-018-0117-3</a>.","apa":"Truckenbrodt, S. M., Sommer, C. M., Rizzoli, S. O., &#38; Danzl, J. G. (2019). A practical guide to optimization in X10 expansion microscopy. <i>Nature Protocols</i>. Nature Publishing Group. <a href=\"https://doi.org/10.1038/s41596-018-0117-3\">https://doi.org/10.1038/s41596-018-0117-3</a>"},"quality_controlled":"1","author":[{"full_name":"Truckenbrodt, Sven M","id":"45812BD4-F248-11E8-B48F-1D18A9856A87","first_name":"Sven M","last_name":"Truckenbrodt"},{"first_name":"Christoph M","last_name":"Sommer","full_name":"Sommer, Christoph M","orcid":"0000-0003-1216-9105","id":"4DF26D8C-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Rizzoli","first_name":"Silvio O","full_name":"Rizzoli, Silvio O"},{"first_name":"Johann G","last_name":"Danzl","full_name":"Danzl, Johann G","orcid":"0000-0001-8559-3973","id":"42EFD3B6-F248-11E8-B48F-1D18A9856A87"}],"year":"2019"},{"pmid":1,"publication":"The EMBO Journal","title":"Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission","_id":"145","oa":1,"ddc":["570"],"abstract":[{"text":"Aged proteins can become hazardous to cellular function, by accumulating molecular damage. This implies that cells should preferentially rely on newly produced ones. We tested this hypothesis in cultured hippocampal neurons, focusing on synaptic transmission. We found that newly synthesized vesicle proteins were incorporated in the actively recycling pool of vesicles responsible for all neurotransmitter release during physiological activity. We observed this for the calcium sensor Synaptotagmin 1, for the neurotransmitter transporter VGAT, and for the fusion protein VAMP2 (Synaptobrevin 2). Metabolic labeling of proteins and visualization by secondary ion mass spectrometry enabled us to query the entire protein makeup of the actively recycling vesicles, which we found to be younger than that of non-recycling vesicles. The young vesicle proteins remained in use for up to ~ 24 h, during which they participated in recycling a few hundred times. They were afterward reluctant to release and were degraded after an additional ~ 24–48 h. We suggest that the recycling pool of synaptic vesicles relies on newly synthesized proteins, while the inactive reserve pool contains older proteins.","lang":"eng"}],"publication_status":"published","citation":{"mla":"Truckenbrodt, Sven M., et al. “Newly Produced Synaptic Vesicle Proteins Are Preferentially Used in Synaptic Transmission.” <i>The EMBO Journal</i>, vol. 37, no. 15, e98044, Wiley, 2018, doi:<a href=\"https://doi.org/10.15252/embj.201798044\">10.15252/embj.201798044</a>.","apa":"Truckenbrodt, S. M., Viplav, A., Jähne, S., Vogts, A., Denker, A., Wildhagen, H., … Rizzoli, S. (2018). Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission. <i>The EMBO Journal</i>. Wiley. <a href=\"https://doi.org/10.15252/embj.201798044\">https://doi.org/10.15252/embj.201798044</a>","ieee":"S. M. Truckenbrodt <i>et al.</i>, “Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission,” <i>The EMBO Journal</i>, vol. 37, no. 15. Wiley, 2018.","ama":"Truckenbrodt SM, Viplav A, Jähne S, et al. Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission. <i>The EMBO Journal</i>. 2018;37(15). doi:<a href=\"https://doi.org/10.15252/embj.201798044\">10.15252/embj.201798044</a>","ista":"Truckenbrodt SM, Viplav A, Jähne S, Vogts A, Denker A, Wildhagen H, Fornasiero E, Rizzoli S. 2018. Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission. The EMBO Journal. 37(15), e98044.","short":"S.M. Truckenbrodt, A. Viplav, S. Jähne, A. Vogts, A. Denker, H. Wildhagen, E. Fornasiero, S. Rizzoli, The EMBO Journal 37 (2018).","chicago":"Truckenbrodt, Sven M, Abhiyan Viplav, Sebsatian Jähne, Angela Vogts, Annette Denker, Hanna Wildhagen, Eugenio Fornasiero, and Silvio Rizzoli. “Newly Produced Synaptic Vesicle Proteins Are Preferentially Used in Synaptic Transmission.” <i>The EMBO Journal</i>. Wiley, 2018. <a href=\"https://doi.org/10.15252/embj.201798044\">https://doi.org/10.15252/embj.201798044</a>."},"quality_controlled":"1","author":[{"full_name":"Truckenbrodt, Sven M","id":"45812BD4-F248-11E8-B48F-1D18A9856A87","first_name":"Sven M","last_name":"Truckenbrodt"},{"full_name":"Viplav, Abhiyan","first_name":"Abhiyan","last_name":"Viplav"},{"last_name":"Jähne","first_name":"Sebsatian","full_name":"Jähne, Sebsatian"},{"last_name":"Vogts","first_name":"Angela","full_name":"Vogts, Angela"},{"last_name":"Denker","first_name":"Annette","full_name":"Denker, Annette"},{"last_name":"Wildhagen","first_name":"Hanna","full_name":"Wildhagen, Hanna"},{"full_name":"Fornasiero, Eugenio","first_name":"Eugenio","last_name":"Fornasiero"},{"last_name":"Rizzoli","first_name":"Silvio","full_name":"Rizzoli, Silvio"}],"tmp":{"image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"year":"2018","intvolume":"        37","isi":1,"publist_id":"7778","month":"08","date_created":"2018-12-11T11:44:52Z","status":"public","article_number":"e98044","volume":37,"issue":"15","file_date_updated":"2020-07-14T12:44:56Z","publisher":"Wiley","article_type":"original","has_accepted_license":"1","department":[{"_id":"JoDa"}],"doi":"10.15252/embj.201798044","language":[{"iso":"eng"}],"acknowledgement":"We thank Reinhard Jahn for providing a plasmid for YFP-SNAP25. We thank Erwin Neher for help with the development of the mathematical model of the synaptic vesicle life cycle. We thank Martin Meschkat, Andreas Höbartner, Annedore Punge, and Peer Hoopmann for help with the experiments. We thank Burkhard Rammner for providing the illustrations of synaptic vesicle and protein dynamics. We thank Manuel Maidorn, Martin Helm, and Katharina N. Richter for critically reading the manuscript. S.T. was supported by an Excellence Stipend of the Göttingen Graduate School for Neurosciences, Biophysics, and Molecular Biosciences (GGNB). E.F.F. is a recipient of long-term fellowships from the European Molecular Biology Organization (ALTF_797-2012) and from the Human Frontier Science Program (HFSP_LT000830/2013). The work was supported by grants to S.O.R. from the European Research Council (ERC-2013-CoG NeuroMolAnatomy) and from the Deutsche Forschungsgemeinschaft (Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, SFB1190/P09, SFB889/A05, and SFB1286/A03, and DFG RI 1967 7/1). The nanoSIMS instrument was funded by the German Federal Ministry of Education and Research (03F0626A).","article_processing_charge":"No","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","oa_version":"Published Version","date_updated":"2023-09-13T09:02:48Z","type":"journal_article","day":"01","scopus_import":"1","publication_identifier":{"issn":["0261-4189"]},"external_id":{"pmid":["29950309"],"isi":["000440416900005"]},"file":[{"file_size":2846470,"creator":"dernst","content_type":"application/pdf","relation":"main_file","date_created":"2018-12-17T14:17:29Z","file_name":"2018_EMBO_Truckenbrodt.pdf","checksum":"a540feb6c9af6aefc78de531461a8835","access_level":"open_access","file_id":"5710","date_updated":"2020-07-14T12:44:56Z"}],"date_published":"2018-08-01T00:00:00Z"},{"issue":"9","volume":19,"publisher":"EMBO","file_date_updated":"2020-07-14T12:47:32Z","isi":1,"intvolume":"        19","article_number":"e45836","status":"public","month":"09","date_created":"2019-05-28T13:16:08Z","quality_controlled":"1","author":[{"id":"45812BD4-F248-11E8-B48F-1D18A9856A87","full_name":"Truckenbrodt, Sven M","first_name":"Sven M","last_name":"Truckenbrodt"},{"last_name":"Maidorn","first_name":"Manuel","full_name":"Maidorn, Manuel"},{"last_name":"Crzan","first_name":"Dagmar","full_name":"Crzan, Dagmar"},{"full_name":"Wildhagen, Hanna","last_name":"Wildhagen","first_name":"Hanna"},{"full_name":"Kabatas, Selda","last_name":"Kabatas","first_name":"Selda"},{"full_name":"Rizzoli, Silvio O","last_name":"Rizzoli","first_name":"Silvio O"}],"citation":{"ista":"Truckenbrodt SM, Maidorn M, Crzan D, Wildhagen H, Kabatas S, Rizzoli SO. 2018. X10 expansion microscopy enables 25‐nm resolution on conventional microscopes. EMBO reports. 19(9), e45836.","chicago":"Truckenbrodt, Sven M, Manuel Maidorn, Dagmar Crzan, Hanna Wildhagen, Selda Kabatas, and Silvio O Rizzoli. “X10 Expansion Microscopy Enables 25‐nm Resolution on Conventional Microscopes.” <i>EMBO Reports</i>. EMBO, 2018. <a href=\"https://doi.org/10.15252/embr.201845836\">https://doi.org/10.15252/embr.201845836</a>.","short":"S.M. Truckenbrodt, M. Maidorn, D. Crzan, H. Wildhagen, S. Kabatas, S.O. Rizzoli, EMBO Reports 19 (2018).","ieee":"S. M. Truckenbrodt, M. Maidorn, D. Crzan, H. Wildhagen, S. Kabatas, and S. O. Rizzoli, “X10 expansion microscopy enables 25‐nm resolution on conventional microscopes,” <i>EMBO reports</i>, vol. 19, no. 9. EMBO, 2018.","ama":"Truckenbrodt SM, Maidorn M, Crzan D, Wildhagen H, Kabatas S, Rizzoli SO. X10 expansion microscopy enables 25‐nm resolution on conventional microscopes. <i>EMBO reports</i>. 2018;19(9). doi:<a href=\"https://doi.org/10.15252/embr.201845836\">10.15252/embr.201845836</a>","apa":"Truckenbrodt, S. M., Maidorn, M., Crzan, D., Wildhagen, H., Kabatas, S., &#38; Rizzoli, S. O. (2018). X10 expansion microscopy enables 25‐nm resolution on conventional microscopes. <i>EMBO Reports</i>. EMBO. <a href=\"https://doi.org/10.15252/embr.201845836\">https://doi.org/10.15252/embr.201845836</a>","mla":"Truckenbrodt, Sven M., et al. “X10 Expansion Microscopy Enables 25‐nm Resolution on Conventional Microscopes.” <i>EMBO Reports</i>, vol. 19, no. 9, e45836, EMBO, 2018, doi:<a href=\"https://doi.org/10.15252/embr.201845836\">10.15252/embr.201845836</a>."},"year":"2018","tmp":{"image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"_id":"6499","oa":1,"ddc":["580"],"title":"X10 expansion microscopy enables 25‐nm resolution on conventional microscopes","publication":"EMBO reports","abstract":[{"text":"Expansion microscopy is a recently introduced imaging technique that achieves super‐resolution through physically expanding the specimen by ~4×, after embedding into a swellable gel. The resolution attained is, correspondingly, approximately fourfold better than the diffraction limit, or ~70 nm. This is a major improvement over conventional microscopy, but still lags behind modern STED or STORM setups, whose resolution can reach 20–30 nm. We addressed this issue here by introducing an improved gel recipe that enables an expansion factor of ~10× in each dimension, which corresponds to an expansion of the sample volume by more than 1,000‐fold. Our protocol, which we termed X10 microscopy, achieves a resolution of 25–30 nm on conventional epifluorescence microscopes. X10 provides multi‐color images similar or even superior to those produced with more challenging methods, such as STED, STORM, and iterative expansion microscopy (iExM). X10 is therefore the cheapest and easiest option for high‐quality super‐resolution imaging currently available. X10 should be usable in any laboratory, irrespective of the machinery owned or of the technical knowledge.","lang":"eng"}],"publication_status":"published","external_id":{"isi":["000443682200009"]},"file":[{"file_id":"6500","date_updated":"2020-07-14T12:47:32Z","checksum":"6ec90abc637f09cca3a7b6424d7e7a26","file_name":"2018_embo_Truckenbrodt.pdf","access_level":"open_access","relation":"main_file","date_created":"2019-05-28T13:17:19Z","creator":"kschuh","content_type":"application/pdf","file_size":2005572}],"date_published":"2018-09-01T00:00:00Z","scopus_import":"1","publication_identifier":{"issn":["1469-221X"],"eissn":["1469-3178"]},"article_processing_charge":"No","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","day":"01","oa_version":"Published Version","type":"journal_article","date_updated":"2023-09-19T14:52:32Z","department":[{"_id":"JoDa"}],"has_accepted_license":"1","doi":"10.15252/embr.201845836","language":[{"iso":"eng"}]}]