Testicular Radiation Damage Analysis

After collection, testicular samples, divided into 4–5 pieces from each group (control, low-, intermediate-, and high-level LDR radiation), were sectioned and immediately fixed in 2.5% glutaraldehyde (Sigma-Aldrich, St. Louis, MO, USA) in PBS, at 4 °C for at least 48 h before processing for LM and TEM as previously described [31 (link),32 (link)]. Semithin sections (1 µm thick) were stained with methylene blue and images were taken using a digital camera (Leica DFC230) mounted on a LM (Zeiss Axioscope, Carl Zeiss Microscopy GmbH, Jena, Germany). Ultrathin sections (60–80 nm) were cut with a diamond knife, mounted on copper grids, and contrasted with UranyLess and Lead citrate (SIC, Rome, Italy). Images were captured using Philips TEM CM100 electron microscopes operating at 80 kV [33 (link),34 (link),35 (link)].