{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T02:07:02Z","timestamp":1791252422839,"version":"4.1.0"},"reference-count":38,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2017,6,12]],"date-time":"2017-06-12T00:00:00Z","timestamp":1497225600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004586","name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Microscopy"],"published-print":{"date-parts":[[2017,9]]},"abstract":"<jats:title>Summary<\/jats:title>\n                  <jats:p>\n                    Biofilms are frequently related to invasive fungal infections and are reported to be more resistant to antifungal drugs than planktonic cells. The structural complexity of the biofilm as well as the presence of a polymeric extracellular matrix (ECM) is thought to be associated with this resistant behavior. Scanning electron microscopy (SEM) after room temperature glutaraldehyde\u2010based fixation, have been used to study fungal biofilm structure and drug susceptibility but they usually fail to preserve the ECM and, therefore, are not an optimised methodology to understand the complexity of the fungal biofilm. Thus, in this work, we propose a comparative analysis of room\u2010temperature and cryofixation\/freeze substitution of\n                    <jats:italic>Candida albicans<\/jats:italic>\n                    biofilms for SEM observation. Our experiments showed that room\u2010temperature fixative protocols using glutaraldehyde and osmium tetroxide prior to alcohol dehydration led to a complete extraction of the polymeric ECM of biofilms. ECM from fixative and alcohol solutions were recovered after all processing steps and these structures were characterised by biochemistry assays, transmission electron microscopy and mass spectrometry. Cryofixation techniques followed by freeze\u2010substitution lead to a great preservation of both ECM structure and\n                    <jats:italic>C. albicans<\/jats:italic>\n                    biofilm cells, allowing the visualisation of a more reliable biofilm structure. These findings reinforce that cryofixation should be the indicated method for SEM sample preparation to study fungal biofilms as it allows the visualisation of the EMC and the exploration of the biofilm structure to its fullest, as its structural\/functional role in interaction with host cells, other pathogens and for drug resistance assays.\n                  <\/jats:p>","DOI":"10.1111\/jmi.12580","type":"journal-article","created":{"date-parts":[[2017,6,12]],"date-time":"2017-06-12T08:56:47Z","timestamp":1497257807000},"page":"409-419","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["<i>Candida albicans<\/i>\n                    biofilms: comparative analysis of room\u2010temperature and cryofixation for scanning electron microscopy"],"prefix":"10.1111","volume":"267","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5600-4367","authenticated-orcid":false,"given":"T.","family":"VILA","sequence":"first","affiliation":[{"name":"Instituto de Biof\u00edsica Carlos Chagas Filho Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B.B.","family":"FONSECA","sequence":"additional","affiliation":[{"name":"Instituto de Biof\u00edsica Carlos Chagas Filho Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.M.L.","family":"DA CUNHA","sequence":"additional","affiliation":[{"name":"N\u00facleo Multidisciplinar de Pesquisa em Biologia UFRJ\u2010Xer\u00e9m\u2010NUMPEX\u2010BIO Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.R.C.","family":"DOS SANTOS","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica M\u00e9dica Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"ISHIDA","sequence":"additional","affiliation":[{"name":"Instituto de Ci\u00eancias Biom\u00e9dicas Universidade de S\u00e3o Paulo  S\u00e3o Paulo Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"BARRETO\u2010BERGTER","sequence":"additional","affiliation":[{"name":"Instituto de Microbiologia Prof. Paulo de G\u00f3es Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W.","family":"DE SOUZA","sequence":"additional","affiliation":[{"name":"Instituto de Biof\u00edsica Carlos Chagas Filho Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"},{"name":"Instituto Nacional de Ci\u00eancia e Tecnologia de Biologia Estrutural e Bioimagem Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"ROZENTAL","sequence":"additional","affiliation":[{"name":"Instituto de Biof\u00edsica Carlos Chagas Filho Universidade Federal do Rio de Janeiro  Rio de Janeiro Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2017,6,12]]},"reference":[{"key":"e_1_2_9_2_1","first-page":"36","article-title":"Image processing with ImageJ","volume":"11","author":"Abramoff M.","year":"2004","journal-title":"Biophoton. 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