Cross beam 1540esb electron microscope

To evaluate three-dimensional shape and features of cells grown on Thermanox™ Coverslips and Elastosil^® Film, we analyzed our samples through scanning electron microscopy (SEM). Samples were fixed in 0.5% glutaraldehyde (Sigma Aldrich) in 0.1 M sodium cacodylate (Sigma Aldrich) for 1 h, then post fixed with osmium tetroxide (Società Italiana Chimici, Roma, Italy) and dehydrated through a series of passages in increasing ethanol baths. Then, cells were dried in pure hexamethyldisilane (HMDS, Fluka Chemie AG, Buchs, Switzerland). At the end, samples were mounted on stubs, coated with gold in a sputter coater (Agar Scientific, Stansted, UK) and then examined on a Cross-Beam 1540EsB electron microscope (Carl Zeiss GmbH, Oberkochen, Germany).

Coated specimens were observed through SEM using secondary electron detection (Cross‐Beam 1540EsB electron microscope (Supra55, Carl Zeiss)). Acceleration voltage was set to 0.4 to 1.5 kV, working distance to 4 to 6 mm and enlargement up to 20 kx. Multiple SEM images at various magnifications were acquired for both studied samples.

For scanning electron microscopy (SEM) observation, cells cultured on the different substrates were fixed in 0.5% glutaraldehyde for 1 h at 4 °C, washed in cacodylate buffer, and then post-fixed with 1% OsO4 for an additional hour. Fixed specimens were dehydrated through a series of passages in increasing ethanol baths and dried in pure hexamethyldisilazane (HMDS, Fluka Chemie AG, Buchs, Switzerland). Finally, samples were mounted on stubs, coated with gold in a sputter coater (Agar Scientific Ltd., Stansted, England) and then examined on a Cross-Beam 1540EsB electron microscope (Carl Zeiss Microscopy, Oberkochen, Germany).