Briefly,
rehydrated sections were incubated in 3% acetic acid (in DI water) for
3 min at RT and then placed directly into 1% Alcian blue 8GX (in 3%
acetic acid, pH 2.5) for an additional 30 min at RT. Sections were then
rinsed in running tap water for 5 min, rinsed in DI water for 1 min,
dehydrated, cleared, and mounted.