Wet-surface-enhanced ellipsometric contrast microscopy identifies slime as a major adhesion factor during bacterial surface motility.
In biology, the extracellular matrix (ECM) promotes both cell adhesion and specific recognition, which is essential for central developmental processes in both eukaryotes and prokaryotes. However, live studies of the dynamic interactions between cells and the ECM, for example during motility, have been greatly impaired by imaging limitations: mostly the ability to observe the ECM at high resolution in absence of specific staining by live microscopy. To solve this problem, we developed a unique technique, wet-surface enhanced ellipsometry contrast (Wet-SEEC), which magnifies the contrast of transparent organic materials deposited on a substrate (called Wet-surf) with exquisite sensitivity. We show that Wet-SEEC allows both the observation of unprocessed nanofilms as low as 0.2 nm thick and their accurate 3D topographic reconstructions, directly by standard light microscopy. We next used Wet-SEEC to image slime secretion, a poorly defined property of many prokaryotic and eukaryotic organisms that move across solid surfaces in absence of obvious extracellular appendages (gliding). Using combined Wet-SEEC and fluorescent-staining experiments, we observed slime deposition by gliding Myxococcus xanthus cells at unprecedented resolution. Altogether, the results revealed that in this bacterium, slime associates preferentially with the outermost components of the motility machinery and promotes its adhesion to the substrate on the ventral side of the cell. Strikingly, analogous roles have been proposed for the extracellular proteoglycans of gliding diatoms and apicomplexa, suggesting that slime deposition is a general means for gliding organisms to adhere and move over surfaces.
Laboratoire de Chimie Bactérienne, Centre National de la Recherche Scientifique Unité Mixte de Recherche 7283, Institut de Microbiologie de la Méditerranée, Aix-Marseille University, 13402 Marseille cedex 20, France.
Adrien
A
Ducret
Marie-Pierre
MP
Valignat
Fabrice
F
Mouhamar
Tâm
T
Mignot
Olivier
O
Theodoly
D001422:Bacterial Adhesion; D005109:Extracellular Matrix; D005456:Fluorescent Dyes; D016941:Myxococcus xanthus / Q000502:physiology*; D013499:Surface Properties; D017155:Wettability
D005456:Fluorescent Dyes