Evo 40 vp sem

Sample processing was performed using glass coverslips precoated with 1 mg/mL poly-l-lysine. Protozoa were fixed for 1 h in 2.5% glutaraldehyde diluted in 0.1 M cacodylate buffer pH 7.2. Cells were subsequently adhered to coverslips, postfixed for 1 h with 1% osmium tetroxide diluted in cacodylate buffer, and dehydrated in a graded alcohol series (50%, 70%, 90%, and two exchanges of 100% ethanol for 10 min each step). Samples were critical-point dried in a Leica EM CPD030 apparatus (Leica, Wetzlar, Germany). Specimens were sputtered with gold in a Balzers FL9496 unit (Postfach 1000 FL-9496 Balzers Liechtenstein) and observed in an EVO 40 VP SEM (Zeiss, Germany). In all assays performed, approximately 500 cells were observed.

Cells were fixed in 2.5% glutaraldehyde diluted in 0.1 M cacodylate buffer (pH 7.2) for 1 h, following washes in the same buffer and then adhered to poly-L-lysine-coated microscope coverslips. After fixation, parasites were post-fixed with 1% osmium tetroxide diluted in cacodylate buffer for 1 h, dehydrated in ethanol (50%, 70%, 90%, and two exchanges of 100%, 10 min in each step), critical point dried in CO₂ by using a Leica EM CPD030 equipament (Leica, Wetzlar, Germany) and ion sputtered in a Balzers FL9496 unit (Postfach 1000 FL-9496 Balzers Liechtenstein). Samples were observed under an EVO 40 VP SEM (Zeiss, Germany).