{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T14:00:19Z","timestamp":1785247219644,"version":"3.55.0"},"reference-count":28,"publisher":"AIP Publishing","issue":"7","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":27,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Schweizerischer Nationalfonds zur F\u00f6rderung der Wissenschaftlichen Forschung","doi-asserted-by":"publisher","award":["TMCG-2_213773"],"award-info":[{"award-number":["TMCG-2_213773"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100015691","name":"Division of Microbiology and Infectious Diseases","doi-asserted-by":"publisher","award":["U54AI170752"],"award-info":[{"award-number":["U54AI170752"]}],"id":[{"id":"10.13039\/100015691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["pubs.aip.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2026,7,1]]},"abstract":"<jats:p>Laser flash melting and revitrification experiments have recently improved the time resolution of cryo-electron microscopy (cryo-EM) to the microsecond timescale, making it fast enough to observe many of the protein motions that are associated with function. The technique has also opened up a new dimension for cryo-EM sample preparation, making it possible to deposit compounds onto a cryo-EM sample while it is frozen, so that upon flash melting, the embedded particles experience an altered environment. For example, we have recently shown that depositing ultrathin silicon dioxide membranes onto a cryo-EM sample causes particles to detach from the interface upon flash melting, removing preferred particle orientation. Here, we describe an apparatus for physical vapor deposition of compounds onto cryo-EM samples, detailing its design and operation. As a demonstration, we determine that the minimum thickness of silicon dioxide sealing membranes in a laser flash melting experiment is just over two monolayers. Moreover, we show that it is possible to deposit reagents onto the cryo-EM and induce mixing during flash melting, which points toward a new strategy for initiating protein dynamics in time resolved experiments. We propose that the design of our instrument can form the basis for an integrated platform for microsecond time-resolved cryo-EM experiments.<\/jats:p>","DOI":"10.1063\/5.0319284","type":"journal-article","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T13:03:02Z","timestamp":1785243782000},"update-policy":"https:\/\/doi.org\/10.1063\/aip-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["An instrument for physical vapor deposition onto cryo-electron microscopy samples for microsecond time-resolved cryo-electron microscopy"],"prefix":"10.1063","volume":"97","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-9452-5285","authenticated-orcid":false,"given":"Wyatt A.","family":"Curtis","sequence":"first","affiliation":[{"name":"Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics , CH-1015 Lausanne,","place":["Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5499-7596","authenticated-orcid":false,"given":"Constantin R.","family":"Kr\u00fcger","sequence":"additional","affiliation":[{"name":"Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics , CH-1015 Lausanne,","place":["Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2116-4178","authenticated-orcid":false,"given":"Axel P.","family":"Tracol Gavard","sequence":"additional","affiliation":[{"name":"Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics , CH-1015 Lausanne,","place":["Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5837-9119","authenticated-orcid":false,"given":"Jakub","family":"Wenz","sequence":"additional","affiliation":[{"name":"Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics , CH-1015 Lausanne,","place":["Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8611-2684","authenticated-orcid":false,"given":"Marcel","family":"Drabbels","sequence":"additional","affiliation":[{"name":"Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics , CH-1015 Lausanne,","place":["Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8869-5999","authenticated-orcid":false,"given":"Ulrich J.","family":"Lorenz","sequence":"additional","affiliation":[{"name":"Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), Laboratory of Molecular Nanodynamics , CH-1015 Lausanne,","place":["Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"317","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"key":"2026072808341710300_c1","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1063\/1.1707630","article-title":"Applications of metallic shadow-casting to microscopy","volume":"17","year":"1946","journal-title":"J.\u00a0Appl. 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