A Hitachi S-4800 field emission scanning electron microscope with the operating voltage of 3 kV was used to obtain cryogenic SEM images of emulsions and bacterial biofilm. Samples were loaded into rivets mounted onto the cryo-SEM sample holder. The samples were then plunged into slushed liquid nitrogen to freeze the sample. This was followed by fracturing at −130 °C using a flat-edge cold knife and sublimation of the solvent at −95 °C for 15 min to etch the sample. The temperature was lowered back to −130 °C, and the sample was then sputtered with a gold–palladium composite at 10 mA for 132 s before imaging.

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