Rtespa 300 silicon probe

The suspension of α-1,3-glucan in 30% DMSO or 30% DMSO alone was placed onto a mica disk and dried in a desiccator overnight. Then, the samples were imaged in NanoScope V AFM (Veeco, Plainview, NY, USA) using the PeakForce QNM operation mode and a RTESPA-300 silicon probe with spring constant 20–80 N/m (Bruker Nano Inc., Billerica, USA). Three fields 1 µm × 1 µm were imaged for each sample. The images were analyzed with NanoScope Analysis software ver. 1.40 (Veeco, Plainview, NY, USA). It was found that α-1,3-glucan intended for administration to the larvae by injection was in the form of spherical nanoparticles with a diameter of about 7 nm (±0.5) (Supplementary Materials, Figure S2).

The log-phase C. albicans cells in ten-fold diluted YPD medium (12 µl; OD₆₀₀ = 0.2) were incubated in the presence of 5 µM AP2 at 37 °C for 1 and 3 h. To the control samples 2 µl of sterile water was added. After washing with pyrogen-free water, the cells were centrifuged (7000× g, 10 min, 4 °C) and finally suspended in 5 µL of pyrogen-free water. The suspensions were applied on mica disks and dried at 28 °C, and the cells were subjected to imaging using a Nanoscope V AFM (Veeco, Plainview, NY, USA; Analytical Laboratory, Faculty of Chemistry, UMCS, Lublin, Poland). The measurements were performed in the PeakForce QNM operation mode using a RTESPA-300 silicon probe with spring constant 20–80N/m (Bruker Nano Inc. Billerica, MA, USA). Three fields (1 µm × 1 µm and 300 nm × 300 nm) were imaged for each sample. The average root-mean square (RMS) roughness values, Young modulus, and adhesion forces were calculated from twenty 200 nm × 200 nm fields measured on each 1 µm × 1 µm image. The nanomechanical properties were analyzed with NanoScope Analysis software ver. 1.40 (Veeco). Three-dimensional (3D) images were obtained using WSxM software (Nanotec, Tres Cantos, Spain).