{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T10:31:32Z","timestamp":1791109892462,"version":"4.1.0"},"reference-count":25,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2013,1,28]],"date-time":"2013-01-28T00:00:00Z","timestamp":1359331200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2013,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time\u2010consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty\u2010two <jats:italic>Candida<\/jats:italic> spp. and four <jats:italic>Cryptococcus neoformans<\/jats:italic> clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. <jats:italic>Candida albicans<\/jats:italic> clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (<jats:italic>chs3\u0394<\/jats:italic>\/<jats:italic>chs3\u0394<\/jats:italic>) and genes encoding predicted GlycosylPhosphatidylInositol (GPI)\u2014anchored proteins (<jats:italic>pga31<\/jats:italic>\u0394\/\u0394 and <jats:italic>pga62<\/jats:italic>\u0394\/\u0394), were used as controls. As expected, the wild\u2010type strain displayed a significant higher chitin content (<jats:italic>P<\/jats:italic> &lt; 0.001) than <jats:italic>chs3\u0394\/chs3\u0394<\/jats:italic> and <jats:italic>pga31<\/jats:italic>\u0394\/\u0394 especially in the presence of caspofungin. <jats:italic>Ca. parapsilosis<\/jats:italic>, <jats:italic>Ca. tropicalis<\/jats:italic>, and <jats:italic>Ca. albicans<\/jats:italic> showed higher cell wall chitin content. Although no relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi. \u00a9 2013 International Society for Advancement of Cytometry<\/jats:p>","DOI":"10.1002\/cyto.a.22250","type":"journal-article","created":{"date-parts":[[2013,1,28]],"date-time":"2013-01-28T19:00:10Z","timestamp":1359399610000},"page":"324-328","source":"Crossref","is-referenced-by-count":58,"title":["Determination of chitin content in fungal cell wall: An alternative flow cytometric method"],"prefix":"10.1002","volume":"83A","author":[{"given":"Sofia","family":"Costa\u2010de\u2010Oliveira","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana P.","family":"Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel M.","family":"Miranda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Salvador","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria M.","family":"Azevedo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carol A.","family":"Munro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ac\u00e1cio G.","family":"Rodrigues","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cid\u00e1lia","family":"Pina\u2010Vaz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,1,28]]},"reference":[{"key":"e_1_2_5_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/yea.320100702"},{"key":"e_1_2_5_3_2","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1128\/EC.3.4.955-965.2004","article-title":"Proteomic analysis of Candida albicans cell walls reveals covalently bound carbohydrate\u2010active enzymes and adhesins","volume":"3","author":"de Groot PW","year":"2004","journal-title":"Eukaryot Cell"},{"key":"e_1_2_5_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/mmy.39.1.41.53"},{"key":"e_1_2_5_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1369-5274(99)80061-7"},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2000.01729.x"},{"key":"e_1_2_5_7_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2007.05588.x"},{"key":"e_1_2_5_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000040"},{"key":"e_1_2_5_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2004.07.001"},{"key":"e_1_2_5_10_2","doi-asserted-by":"publisher","DOI":"10.1080\/15572536.2006.11832788"},{"key":"e_1_2_5_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/EC.2.1.49-61.2003"},{"key":"e_1_2_5_12_2","first-page":"616","article-title":"Rapid detection of fungi in tissues using calcofluor white and fluorescence microscopy","volume":"108","author":"Monheit JE","year":"1984","journal-title":"Arch Pathol Lab Med"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/EC.4.10.1746-1754.2005"},{"key":"e_1_2_5_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00399-06"},{"key":"e_1_2_5_15_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.00085"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.105.054767"},{"key":"e_1_2_5_17_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dki264"},{"key":"e_1_2_5_18_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1198-743x.2001.00307.x"},{"key":"e_1_2_5_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.43.9.4674-4679.2005"},{"key":"e_1_2_5_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/134.3.717"},{"key":"e_1_2_5_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.23.10570"},{"key":"e_1_2_5_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fgb.2008.08.003"},{"key":"e_1_2_5_23_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20092"},{"key":"e_1_2_5_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00095-07"},{"key":"e_1_2_5_25_2","volume-title":"Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts","author":"CLSI. 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