I am investigating on mitral valve by Movat pentachrome staining, but the results were unsatisfied. Blue (for mucin) and yellow (for collagen) was absent.

Q1: I found at least four companies sold mechanical kits online, including Solarbio (a Chinese company, from which i bought the kit), Abcam, Morphisto (inaccessible protocol) and Newcomer supply. However, protocols varies from company to company. So, which one should I choose? OR, is there any better protocol?

Q2: Solarbio's protocol was instructed as :

1. Routinely deparaffinizing sections with xylene and hydrate through a series of ethanol.

2. Bouin solution, room temperature, overnight.

3. Rinse with running water for 10min.

4. Hyposulfite solution for 5min, and rinse with distilled water for 2 ~ 3 times.

5. Alcian blue for 20min and washed under running water for 2 ~ 5min.

6. Preheated (45~60℃) alkaline alcohol solution for 10 min.

7. Rinse with running water for 2 to 5 minutes.

8. Prepare Weigert hematoxylin stain solution: Solution A: B: C=3:2:1, unknown component.

9. Weigert hematoxylin stain solution for 60min, in the dark place.

10. Rinse with running water and distilled water 2 to 3 times, 3 to 5 minutes each time.

11. Saffron-fuchsia stain solution (prepared by A: B=4:1, unknown component) for 1 min, in the dark place.

12. Rinse with distilled water 2 to 3 times, 3 to 5 minutes each time.

13. Phosphotungstic acid solution for 5min and transferred directly into the weak acid differentiation solution for 5min.

14. Rinse with distilled water 2 to 3 times, 3 to 5 minutes each time.

15. Saffron stain for 5min.

After Step 9, the sections background was stained dark and hard to wash out with distilled water, causing the following staining procedures failed. If I wash it with hydrochloric acid ethanol , the background was clear, yet sections were eventually stained without blue and yellow, but merely red or intense red. So, should I use hydrochloric acid ethanol or just use distilled water with a longer time?