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. 2022 Dec 1;12:1033698. doi: 10.3389/fcimb.2022.1033698

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Copyright © 2022 Morais, Santos, Costa, Martins, Leitão, de Melo Pacífico, Quesada-Gómez, Castelo Branco, Ferreira and Brito

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Confocal laser scanning microscopy analysis (CLSM) of 72h biofilms formed by Clostridioides difficile. Biofilm’s matrix formation by strain ATCC 700057, ICC-45, and epidemic strains LIBA 5756, R20291, was analyzed by FilmTracer™ SYPRO™ Ruby Biofilm Matrix Stain. (A) 2 D and 2.5 D Representative CLSM images of C. difficile biofilms (B) Biofilm thickness in micrometers (C) Mean fluorescence relative units of biofilm matrix formation * represents significant differences among strains (P < 0.05). Scale 20 µm. (Kruskal–Wallis test followed by Dunn’s multiple comparisons test).