Supra 50 vp fesem

Surface structure topography of the scaffolds was examined by a scanning electron microscope at magnifications of × 500 at the Center of Microscopy and Image Analysis, University of Zurich (scanning electron microscopy (SEM); Carl Zeiss Supra 50 VP FESEM, Carl Zeiss). For this purpose, the samples were fixed for 24 h in 2.5% glutaraldehyde solution. Afterward, the scaffolds were rinsed with PBS and dehydrated in ascending concentrations of alcohol (50, 70, 80, and 90%) twice for 15 min. Finally, the scaffolds were immersed three times for 15 min in 94% and 60 min in 100% ethanol. Samples were then subjected to critical point drying (Bal-Tec CPD030), mounted on SEM mounts (Bal-Tec AG, Blazers, Liechtenstein), and were gold sputtered (Balzers SCD 030, Balzers Union, Balzers, Liechtenstein) for 60 s in an argon gas atmosphere at a target distance of 50 mm, at pressure of 5 Pascal (Pa) at 45 mA. SEM images were taken at a working distance of an acceleration voltage of 10 kV.

Finally, the degraded and perfused membranes were visually assessed using scanning electron microscopy (SEM), in order to evaluate the surface structure alterations due to collagen degradation and sperm cell penetration at the different time periods.
Samples were fixed for 24 h in 2.5% glutaraldehyde solution, then rinsed with PBS and dehydrated twice for 15 min in ascending concentrations of ethanol (50%, 70%, 80%, and 90%, respectively). The membranes were then immersed three times for 15 min in 94% and 60 min in 100% ethanol, respectively. Finally, samples were subjected to critical point drying (Bal-Tec CPD030, Balzers, Liechtenstein). All the samples were then cut in the middle and mounted on SEM mounts (Bal-Tec AG, Blazers, Liechtenstein), with each cut shown on the side of the membrane (i.e., upside-down and vice versa). Samples were gold sputtered (Balzers SCD 030, Balzers Union, Balzers, Liechtenstein) for 60 s in an argon gas atmosphere at a target distance of 50 mm and pressure of 5 Pascal (Pa) at 45 mA. SEM images (Supra 50 VP FESEM, Carl Zeiss, Oberkochen, Germany) were taken at a working distance of 9.2 mm, an acceleration voltage of 10 kV, and a magnification of 5000×.