Megaview 2 camera

For immunolocalization, the nettle stem sections were collected on gold grids and blocked for 20 min with normal goat serum (NGS) diluted 1:30 in dilution buffer (0.05 M Tris-HCl pH 7.6, 0.9% w/v NaCl, and 0. 2% w/v BSA). Sections were incubated for 4 h at room temperature with primary antibodies. Antibodies were diluted 1:5 with dilution buffer. Sections were then washed for 20 min in the dilution buffer with 0.1% (v/v) Tween 20 and incubated for 45 min at room temperature with either secondary anti-rat antibodies (for LM5, LM16, JIM5, JIM7, LM14, and LM20) or anti-mouse antibodies (for RU1 and RU2) conjugated to 10 nm gold particles and diluted 1:20 in 0.02 M Tris-HCl pH 8.2. All sections were examined with the Philips MORGAGNI 268 80-kV transmission electron microscope, equipped with MEGAview II camera, and were processed with the Analysis software. Controls were carried out by omitting the primary antibodies; in any case, we have never detected signals due to secondary antibodies. Examples of control images are present as Supplementary Fig. S1.

A Philips CM-10 transmission electron microscope supplied with a Megaview-II camera was used to take the TEM micrographs of magnetite nanoparticles loaded with 0.4 mmol/g CSA. The accelerating voltage of 100 kV was applied; the maximum resolution of the instrument is 0.2 nm. One drop of the highly diluted sol was dried on to a Formwar-coated copper. The average size distribution was determined by evaluating 100 particles using the JMicroVision 1.2.7 software.

C. albicans strain CAF2-1 was grown in YNB liquid cultures for 2 h at 37°C (in 15-ml plastic tubes). Next, miconazole (1 mg/ml in DMSO) was added at a concentration of 10 μg/ml, and the cultures were grown for 18 h to evaluate the cytotoxic effect of this commercial product on the yeast strain. The cytotoxic effect of tomatidine was evaluated by use of the same experiment, except that this compound (1 mg/ml in DMSO) was added at a concentration of 20 μM and the cultures were grown for 18 h. Cells were prepared as described in reference 12 (link). Thin sections were observed with a transmission electron microscope (Philips CM10) with a Mega View II camera. Control cells were obtained in the same way but did not receive drug treatment.