Method

46 steel plates were colonized with biofilm forming S. epidermidis ATCC 35984 in TSB with 1% glucose aerobically at 37°C for 24 hours. Plates were cleansed for non-adherent bacteria before microscopy. Biofilm embedded bacteria were stained with LIVE/DEAD ™ BacLight ™ Bacterial Viability Kit for microscopy and visualized under vital conditions using EVOS™ FL Auto 2 Imaging System (epifluorescence) and an inverse confocal laser scanning microscope LSM510 (CLSM). All steel plates were subjected to epifluorescence microscopy before and after sonication. CLSM and SEM were used to confirm the presence of biofilm embedded bacteria after sonication. Pictures from epifluorescence microscopy were processed for image analysis with help of a macro application (Fiji) and the data was expressed as biofilm coverage rate (BCR). The sonication was performed using a BactoSonic® Bandolin sonicator and the applied effect in each glass test tube (40 kHz, 800W) was measured with a Bruel og Kjær 8103 hydrophone. The amount of bacteria in the sonication fluid was quantified by counting the number of colony forming units (CFU).

Three steel plates acted as negative controls.