Ultra plus field emission microscope

Cells from the control C. albicans culture and those incubated with AAF were centrifuged at 6000×g for 10 min. The supernatant was then discarded and 200 µL of GH solution (glucose, Na-HEPES, sterile water) were added to the dense cell suspension. The samples were re-centrifuged in the same conditions, and the supernatant was discarded again. The fungal cells in a small amount of the GH solution were applied to the transfer and placed in the sublimation chamber. The cooling process was carried out for 12 min at − 92 °C. Then, the samples were transferred to the preparation chamber, where they were broken using a special blade. C. albicans cells were then imaged with a scanning electron microscope (ZEISS Ultra Plus Field Emission microscope). The images were taken at a magnification of 30.000 × and at electron high tension (EHT) 5 kV.

The control culture cells and cells treated with Venetin-1 at the concentration of 100 µg mL⁻¹ were centrifuged (room temperature, 6000×g, 10 min) and the supernatant was discarded. The pellet was suspended in 200 µL of a GH solution (composed of glucose, HEPES, and sterile water) and centrifuged again. Then, the supernatant was removed and 20 µL of the GH solution was added. Next, the cells were transferred into a sublimation chamber with a temperature of − 92 °C for 12 min. After that, the frozen cultures were cut in the preparation chamber and analyzed with a ZEISS Ultra Plus Field Emission Microscope (Carl Zeiss, Germany) with electron high tension (EHT) 5 kV and under magnification 30.000 × ^{39 (link),42 (link)}.