Comparison of SEM and VPSEM imaging techniques with respect to Streptococcus mutans biofilm topography. Issue 2 (2nd December 2013)

• Record Type: Journal Article
• Authors: Weber, Kathryn; Delben, Juliana; Bromage, Timothy G.; Duarte, Simone
• Is Part Of: FEMS microbiology letters. Volume 350:Issue 2(2014:Jan.)
• Page Start: 175
• Page End: 179
• Publication Date: 2013-12-02
• Subjects: Microbiology -- Periodicals; 579
• DOI: 10.1111/1574-6968.12334
• Languages: English
• ISSNs: 0378-1097

Abstract

<abstract abstract-type="main" id="fml12334-abs-0001"> <title>Abstract</title> <p>The study compared images of mature <italic>Streptococcus mutans</italic> biofilms captured at increasing magnification to determine which microscopy method is most acceptable for imaging the biofilm topography and the extracellular polymeric substance (EPS). <italic>In vitro S. mutans</italic> biofilms were imaged using (1) scanning electron microscopy (SEM), which requires a dehydration process; (2) SEM and ruthenium red (SEM‐RR), which has been shown to support the EPS of biofilms during the SEM dehydration; and (3) variable pressure scanning electron microscopy (VPSEM), which does not require the intensive dehydration process of SEM. The dehydration process and high chamber vacuum of both SEM techniques devastated the biofilm EPS, removed supporting structures, and caused cracking on the biofilm surface. The VPSEM offered the most comprehensive representation of the <italic>S. mutans</italic> biofilm morphology. VPSEM provides similar contrast and focus as the SEM, but the procedure is far less time‐consuming, and the use of hazardous chemicals associated with SEM dehydration protocol is avoided with the VPSEM. The inaccurate representations of the biofilm EPS in SEM experimentation is a possible source of inaccurate data and impediments in the study of <italic>S. mutans</italic> biofilms.</p> </abstract>