SEM Imaging of Tannerella forsythia

T. forsythia wild-type and T. forsythia ∆ampG were cultivated with free MurNAc or E. coli PGN, as described above, and samples were prepared for SEM following a published protocol [38 (link)]. In short, 1 ml of each culture was harvested by centrifugation (8000 g, 7 min), cell pellets were washed with phosphate buffered saline and applied to increasing concentrations of ethanol from 25 to 100% in PBS, with incubation of 7 min and removal of ethanol by centrifugation after each step. Samples were sputter-coated with gold (EM SDC005 apparatus; Leica, Wetzlar, Germany) and imaged with an Inspect S50 scanning electron microscope (FEI, Eindhoven, The Netherlands).