It depend on the stains you use. 10 min. maximum for H. then check it under microscope if the stain so dark blue you can use glacial acetic acid to remove the excess of H.
And 4 min. maximum for E. but you should wash slides in tap water very quick.
Here is my protocol. It works in paraffinsections (5-10µm) as well as in mounted frozen sections (10-30 µm). Dewaxed and defatted sections rinse in destilld water.
1. Mayer's Haematoxyline (0.1%) - 5-10 min; 2. rinse well in destilled water; 3. blue in running tape water for 10 min., 4. 1% alcoholic Eosin- solution 1-5 min,; 5. rinse in destiiled water; 6.diffenciate in 70% ETOH (check under the microscope) and 2x 100% ETOH (10 dipps each alcohol); 7. Isopropanol 2 min; 2 x Xylene 5-10 min each; 8. mount in Depex or another xylene soluble mounting medium. Start with a test slide to optimize your stainig times an control after every stainig step the quality under the microscope. Good luck!
it depends on the age of your stain solutions, the older it gets the longer you should stain with. For cryo sections up to only a few minutes, but for paraffin sections normally 10 min for hema, and 4 min for eosin
I think 7-8 mins for haematoxylin and 3-5 min for eosin. To remove excess stain, you can dip the slide into glacial acetic acid mixed ( few ml) in 95% ethanol in dish before mounting.
My routine work, after diparaffinaztion in xylene and rehydrate in different concentate of ethanol,8 min in H, 10 min running tap water and 4 min in eosin