Preparation of Bacterial Samples for SEM

Petri dish microcultures of the strain ENCB-J15 were inoculated, grown in squares of GAE solid medium covered by a circular coverslip, and incubated at 28 °C for 10 days. The coverslip with a culture sample was placed on a 12-well microplate and fixed with 2% glutaraldehyde for 2 h, dehydrated with 10, 20, 30, 40, 50, 60, 70, 80, and 90% ethanol each for 10 min for each dissolution, and finally dehydrated with absolute alcohol for 20 min [27 (link)]. The sample was critically dried with CO₂ using the Emitech K850 Critical Point Dryer to remove all moisture and subsequently mounted on aluminum sample holders with adhesive carbon tape. The sample was coated with a layer of gold at 20 mA for 2 min using Quorum Q15OR-ES equipment. Finally, the sample was observed in a Hitachi SU1510 SEM at 10–15 kW [28 (link)].