{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,14]],"date-time":"2026-06-14T15:52:28Z","timestamp":1781452348235,"version":"3.54.1"},"reference-count":33,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T00:00:00Z","timestamp":1779321600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T00:00:00Z","timestamp":1779321600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100000266","name":"Engineering and Physical Sciences Research Council","doi-asserted-by":"publisher","award":["EP\/X022609\/1"],"award-info":[{"award-number":["EP\/X022609\/1"]}],"id":[{"id":"10.13039\/501100000266","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Dragonfly\u2010 and cicada\u2010wing\u2010inspired titanium nanostructured surfaces exhibit promising bactericidal properties. However, direct visualization of the bacteria\u2013material interface under hydrated conditions remains limited, restricting experimental interrogation of how bacteria interact with nanostructured surfaces. This limitation reflects a long\u2010standing methodological gap, as visualization of bacteria\u2013nanostructure interfaces has largely relied upon indirect or dehydrated imaging approaches. Cryo\u2010electron tomography (cryo\u2010ET) enables 3D visualization of cellular ultrastructure in a native hydrated environment, but imaging bacteria attached to metallic nanostructures by cryo\u2010ET requires preparation of thin electron\u2010transparent lamellae. Obtaining such lamellae from vitrified bacteria on titanium substrates is technically challenging because cryo\u2010focused ion beam (cryo\u2010FIB) milling must simultaneously section soft biological material and hard metal whilst bacteria embedded in vitreous ice are not directly visible. Here, we establish a correlative cryogenic workflow enabling targeted extraction and cryo\u2010ET imaging of defined bacterium\u2013nanopillar interfaces. Individual bacteria interacting with titanium nanopillars are identified by correlative cryo\u2010fluorescence microscopy, followed by targeted cryo\u2010FIB lift\u2010out, transfer to a receiver grid, and thinning into electron\u2010transparent lamellae. The data presented demonstrate that bacteria\u2013nanostructure interfaces can be targeted, extracted, and structurally analysed in situ under fully hydrated conditions. This workflow provides a methodological framework for future cryo\u2010ET studies of bacteria\u2013nanotopography interactions.<\/jats:p>","DOI":"10.1002\/smtd.70715","type":"journal-article","created":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T04:52:39Z","timestamp":1779425559000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Cryo\u2010FIB Lift\u2010Out and Electron Tomography Workflow for Bacteria\u2013Nanopillar Interface Imaging Under Native Conditions: Investigating Dragonfly Inspired Bactericidal Titanium Surfaces"],"prefix":"10.1002","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0688-4260","authenticated-orcid":false,"given":"Chaturanga D.","family":"Bandara","sequence":"first","affiliation":[{"name":"Bristol Dental School Research Laboratories University of Bristol Dorothy Hodgkin Building  Whitson Street Bristol BS1 3NY UK"},{"name":"School of Biochemistry University of Bristol  Bristol UK"},{"name":"Potsdam Imaging and Spectral Analysis Facility (PISA) Helmholtz Centre for Geosciences (GFZ)  Potsdam DE Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6103-4314","authenticated-orcid":false,"given":"Dominik","family":"Pinkas","sequence":"additional","affiliation":[{"name":"Electron Microscopy Core Facility Institute of Molecular Genetics ASCR  Prague Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6347-6167","authenticated-orcid":false,"given":"Martina","family":"Zanova","sequence":"additional","affiliation":[{"name":"Tescan Group a.s.  Brno Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-2646-1765","authenticated-orcid":false,"given":"Martin","family":"Uher","sequence":"additional","affiliation":[{"name":"Tescan Group a.s.  Brno Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9061-8933","authenticated-orcid":false,"given":"Judith","family":"Mantell","sequence":"additional","affiliation":[{"name":"School of Biochemistry University of Bristol  Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1063-8182","authenticated-orcid":false,"given":"Bo","family":"Su","sequence":"additional","affiliation":[{"name":"Bristol Dental School Research Laboratories University of Bristol Dorothy Hodgkin Building  Whitson Street Bristol BS1 3NY UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3813-4410","authenticated-orcid":false,"given":"Angela H.","family":"Nobbs","sequence":"additional","affiliation":[{"name":"Bristol Dental School Research Laboratories University of Bristol Dorothy Hodgkin Building  Whitson Street Bristol BS1 3NY UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2497-1026","authenticated-orcid":false,"given":"Paul","family":"Verkade","sequence":"additional","affiliation":[{"name":"School of Biochemistry University of Bristol  Bristol UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,5,21]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.4c15671"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep16817"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcis.2017.07.021"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2013.01.017"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsami.6b13666"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsbiomaterials.9b01973"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/smll.201200528"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsabm.1c00839"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev\u2010food\u2010060721\u2010022330"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsomega.2c08068"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/adhm.202503902"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.colcom.2025.100863"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.actbio.2018.09.059"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2021.102818"},{"key":"e_1_2_10_16_1","unstructured":"I.MudiyanselageandC. C.Devapriya Bandara \u201cCharacterisation of the Bactericidal Efficacy of Natural Nano\u2010topography Using Dragonfly Wing as a Model \u201dPhD thesis(Queensland University of Technology 2017)."},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/admi.202200608"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/smll.202310149"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/smll.202400784"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1042\/bst20240173"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2018.06.007"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010020\u201015471\u2010x"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.52286"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2005.07.007"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2016.06.007"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102818.2004.01260.x"},{"key":"e_1_2_10_27_1","first-page":"671","volume-title":"Methods in Cell Biology","author":"McDonald K.","year":"2010"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00418\u2010008\u20100500\u20101"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.185.20.6112\u20106118.2003"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010023\u201001861\u20108"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1155\/2021\/8842777"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ultramic.2015.11.001"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1101\/2023.07.11.548578"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010023\u201002113\u20105"}],"container-title":["Small Methods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/smtd.70715","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/smtd.70715","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/smtd.70715","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,14]],"date-time":"2026-06-14T15:01:13Z","timestamp":1781449273000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/smtd.70715"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,21]]},"references-count":33,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2026,6]]}},"alternative-id":["10.1002\/smtd.70715"],"URL":"https:\/\/doi.org\/10.1002\/smtd.70715","archive":["Portico"],"relation":{},"ISSN":["2366-9608","2366-9608"],"issn-type":[{"value":"2366-9608","type":"print"},{"value":"2366-9608","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,21]]},"assertion":[{"value":"2026-02-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-07","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70715"}}