Open AccessComparison of SEM and VPSEM imaging techniques with respect to S treptococcus mutans biofilm topography
Author(s) -
Weber Kathryn,
Delben Juliana,
Bromage Timothy G.,
Duarte Simone
Publication year - 2014
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12334
Subject(s) - biofilm , scanning electron microscope , extracellular polymeric substance , dehydration , streptococcus mutans , environmental scanning electron microscope , chemistry , microbiology and biotechnology , materials science , biomedical engineering , chemical engineering , bacteria , biology , composite material , biochemistry , medicine , genetics , engineering
The study compared images of mature S treptococcus mutans biofilms captured at increasing magnification to determine which microscopy method is most acceptable for imaging the biofilm topography and the extracellular polymeric substance ( EPS ). In vitro S . mutans 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 S . mutans 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 S . mutans biofilms.