For the environmental scanning electron microscopy (ESEM) the samples were fixed by dehydration with ethanol. ESEM operates in a low vacuum mode, and this condition improves the images resolution. In order to prepare the sample for the analysis, the culture medium was removed and cells were incubated overnight at 4°C with a solution composed of 1.5% (v/v) glutaraldehyde 50% (v/v) in water and 0.1 M sodium cacodylate (pH=7.1-7.2). Then, samples were rinsed with 0.1 M sodium cacodylate buffer and incubated for 5 min each with increasing ethanol concentrations (20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 96% and 100% v/v). This passage was repeated twice. The images were acquired using the ESEM ZEISS EVO 50 EP.

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