TY - JOUR
T1 - Extracellular matrix reorganization during cryo preparation for scanning electron microscope imaging of staphylococcus aureus biofilms
AU - Wu, Yong
AU - Liang, Jing
AU - Rensing, Kim
AU - Chou, Tseng Ming
AU - Libera, Matthew
N1 - Publisher Copyright:
© Microscopy Society of America 2014.
PY - 2014/4/11
Y1 - 2014/4/11
N2 - Biofilms are three-dimensional communities of bacteria distributed in a highly hydrated extracellular matrix (ECM). They can be visualized by scanning electron microscopy (SEM), but the requisite SEM sample preparation can modify the biofilm morphology. Here, four different approaches to prepare biofilms of hydrated Staphylococcus aureus for SEM imaging are compared. In order of increasing cooling effectiveness these are: (1) drying in air; (2) plunging in liquid nitrogen; (3) plunging in liquid ethane; and (4) high pressure freezing with liquid nitrogen. These different methods give rise to markedly different biofilm morphologies, which are revealed by cryo-SEM imaging. Significantly, high-pressure frozen biofilms exhibit a rich network of nanoscale ECM fibers surrounding individual bacteria throughout the biofilm thickness. This structure is entirely lost when similar biofilms are dried in air, and it is substantially modified when these biofilms are plunged into liquid nitrogen or liquid ethane.
AB - Biofilms are three-dimensional communities of bacteria distributed in a highly hydrated extracellular matrix (ECM). They can be visualized by scanning electron microscopy (SEM), but the requisite SEM sample preparation can modify the biofilm morphology. Here, four different approaches to prepare biofilms of hydrated Staphylococcus aureus for SEM imaging are compared. In order of increasing cooling effectiveness these are: (1) drying in air; (2) plunging in liquid nitrogen; (3) plunging in liquid ethane; and (4) high pressure freezing with liquid nitrogen. These different methods give rise to markedly different biofilm morphologies, which are revealed by cryo-SEM imaging. Significantly, high-pressure frozen biofilms exhibit a rich network of nanoscale ECM fibers surrounding individual bacteria throughout the biofilm thickness. This structure is entirely lost when similar biofilms are dried in air, and it is substantially modified when these biofilms are plunged into liquid nitrogen or liquid ethane.
KW - S. aureus
KW - biofilm
KW - cryo
KW - extracellular matrix
KW - scanning electron microscopy
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U2 - 10.1017/S143192761401277X
DO - 10.1017/S143192761401277X
M3 - Article
C2 - 24994510
AN - SCOPUS:84911991609
SN - 1431-9276
VL - 20
SP - 1348
EP - 1355
JO - Microscopy and Microanalysis
JF - Microscopy and Microanalysis
IS - 5
ER -