2.6.6. Histological Analysis

Samples were collected at the end of the experiment and processed for histology. Samples were then fixed in 10% neutral-buffered formalin and then kept at 4 ◦C until the staining procedures. Alcian Blue staining was performed by rinsing the samples in 3% acetic acid, keeping them in 1% alcian blue solution (Sigma, A-3157) for 1 h. After that, the stain was poured off and sections were washed with water, let to dry, and then rinsed in absolute alcohol, cleared in xylene, and mounted in Entellan rapid (Merck Millipore, Darmstadt, Germany, 1.07960.0500).
