Multimode 8 nanoscope afm equipment

A. baumannii bacteria and vesicles were imaged by AFM as described previously (26 (link)) with slight modifications. Briefly, 10 µL of bacterial samples collected from the logarithmic phase of growth with different serial dilutions were placed onto freshly cleaved mica (Goodfellow Cambridge Ltd., Cambridge, United Kingdom). The samples were blot dried and desiccated prior to imaging. Imaging was done on a Multimode 8 Nanoscope AFM equipment (Digital Instruments, Santa Barbara) using tapping mode TM. A silicon probe was oscillated at its resonant frequency of approximately 300 kHz, selected by the Nanoscope software. Images were collected in the air at a scan rate of 0.8–1.5 Hz, depending on the size of the scan and the number of samples (256 or 512 samples/image). The final images were plane fitted in both axes and presented in a surface plot of the height mode.

The OMVs were imaged by Atomic Force Microscopy (AFM) as described previously [18 (link)]. Briefly, 10 μl of the OMV samples were placed onto freshly cleaved mica (Goodfellow Cambridge Ltd., Cambridge, United Kingdom). The samples were blot dried and desiccated prior to imaging. Imaging was done on a Multimode 8 Nanoscope AFM equipment (Digital Instruments, Santa Barbara) using Tapping ModeTM. A silicon probe was oscillated at its resonant frequency of approximately 300 kHz, selected by the Nanoscope software. Images were collected in air at a scan rate of 0.8–1.5 Hz, depending on the size of the scan and the number of samples (256 or 512 samples/image). The final images were plane fitted in both axes and presented in a surface plot of the height mode.