[Deparaffinization]
(1) Xylene down I: 10 min
(2) Xylene down II: 10 min
(3) 100% ethyl alcohol (= ethanol) down I: 10 min
(4) 100% ethanol down II: 10 min
(5) 90% ethanol down: 10 min
(6) 70% ethanol down: 10 min
(7) Distilled water (= D.W.): 5 min

[Staining]
(1) 3% CH₃COOH solution (1): 2 min
(2) pH 2.5 alcian blue: 1 h
(3) D.W.: 30 sec
(4) 3% CH₃COOH solution (2): 30 sec
(5) D.W.: 30 sec
(6) 0.3% Na₂CO₃ solution: 20 min
(7) D.W.: 30 sec
(8) 0.5% HIO₄ solution: 7 min
(9) Washing with running tap water: 5 min
(10) D.W.: 30 sec
(11) Schiff's reagent: 10 min
(12) Washing with NaHSO₃ solution (1): 3 min
(13) Washing with NaHSO₃ solution (2): 3 min
(14) Washing with NaHSO₃ solution (3): 3 min
(15) Washing with running tap water: 5 min
(16) D.W.: 30 sec

[Dehydration and Immersion]
(1) 70% ethanol up: 10 min
(2) 90% ethanol up: 2 min
(3) 100% ethanol up I: 3 min
(4) 100% ethanol up II: 5 min
(5) Creosote-xylene up: 5 min
(6) Xylene up I: 5 min
(7) Xylene up II: 10 min
(8) Mount of sections in 6-7 drops of a mixed solution of Canada balsam and xylene with a 24 x 45-55 mm micro cover glass

[Components of each solution]
(1) pH 2.5 alcian blue (200 ml): Add glacial acetic acid (6 ml) to D.W. (194 ml), and then dissolve alcian blue 8GX (0.2 g) in it. After adjusting this solution to pH 2.5 with sodium phosphate-buffer, add a small amount (to a pick of earwax) of powdery thymol (i.e., antiseptic) to it
(2) Sodium phosphate-buffer: Dissolve Na₂HPO₄.12H₂O (7.16 g) in D.W. (100 ml)
(3) Schiff's reagent (200 ml): Boil D.W. (200 ml). After being fire off, add "basic fuchsine" (= parafuchsine: 1 g) to this D.W., in which basic fuchsine is gradually dissoluble. Further boil this solution for 5-10 min to dissolve basic fuchsine entirely. After chilling this solution until 50 C at a room temperature, filtrate it. Add 1N-HCl (20 ml) to this solution, and then mix it by stirring to dissolve some drifting matters. After chilling this solution until 25 C with running tap water, dissolve NaHSO₃ (1 g) in it. Reserve this solution in a refrigerator for 2-3 days (solution coloring turns "orange") before using it (this solution needs a screen for light-interception). Need renewal of this solution if its coloring turned a tinge of red
(4) 1N-HCl (100 ml): Add 37.2% HCl (8.35 ml) to some amount of D.W., and then increase D.W. up to 100 ml
(5) NaHSO₃ solution (600 ml): Mix 10% NaHSO₃ (36 ml) with 1N-HCl (30 ml), and then add D.W. to this solution (up to 600 ml). Make the NaHSO₃ solution just before using it
(6) Creosote-xylene (200 ml): Mix creosote (50 ml) with xylene (150 ml)