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Observation of the mucin beads by scanning electron microscopy (SEM)

RG Raphaële Gresse
FC Frédérique Chaucheyras-Durand
SD Sylvain Denis
MB Martin Beaumont
TW Tom Van de Wiele
EF Evelyne Forano
SB Stéphanie Blanquet-Diot
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To control that microorganisms were able to adhere to the bead surface during the fermentation process, some beads were sampled after a 48-h stay in the mucin bead compartment during a control assay. Beads were gently washed 3 times in sterile PBS 1×. Then, they were fixated by immersion at 4 °C overnight in a 0.2-mol/L sodium cacodylate (pH 7.4) buffer containing 2% glutaraldehyde. After fixation, mucin beads were dehydrated using successive 20 min incubations in ethanol 70%, 95% and 100%. A final dehydration step included 20 min incubation in a mix of ethanol 100% and hexamethyldisilazane. Samples were dried overnight under a chamber and stained with gold-palladium deposition. SEM images were taken with a JEOL 6060 Low vacuum microscope (Jeol Europe SAS, Croissy sur Seine, France) at the Centre Imagerie Cellulaire Santé (CICS) from Clermont-Auvergne University.

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