“…dehydrated with series of ethanol to DW, immersed in toluidine blue solution for 30 s, washed 3 times with DW for 3 min each time, differentiated with 95% ethanol for 10 min, dehydrated with 100% ethanol, and cleared in xylene and mounted with neutral gum.PAS staining: the paraffin sections were dewaxed and dehydrated with series of ethanol to DW, oxidized with 0.5% periodic acid solution for 5 min, washed with DW, incubated with Schiff's solution at 37°C for 30 min, washed with sodium sulphite solution 2-3 times for 1 min each time, washed with DW, counterstained with haematoxylin, differentiated by 0.5% hydrochloric ethanol, ascending dehydrated with gradient ethanol, and cleared in xylene and mounted with neutral gum(Kandyel, Abumandour, et al, 2021b).Alcian blue-periodic acid Schiff's staining: the paraffin sections were dewaxed and dehydrated with series of ethanol to DW, immersed in Alcian blue solution for 30 min, washed with DW for 3 min, immersed in 1% periodic acid solution for 5 min, washed with DW for 3 min, incubated with Schiff's solution at 37°C for 30 min, differentiated by 2.5% sodium metabisulphite for 5 s, washed with DW, ascending dehydration with gradient ethanol, and cleared in xylene and mounted with neutral gum.High iron diamine-Alcian blue staining: the paraffin sections were dewaxed and dehydrated with series of ethanol to DW, incubated in high iron diamine solution at room temperature for 24 h, washed with DW, incubated in Alcian blue solution for 20 min, washed with DW, ascending dehydration with gradient ethanol, and cleared in xylene and mounted with neutral gum.…”