File talk:Heart histology 001.jpg
From Embryology
Staining solutions
A. Hematoxylin (CI No. 75290), 0.5 gm; ferric ammonium sulfate, 1.0 gm; 0.6 N H$Ol (aq.), 50 ml; 95% ethanol, 50 ml. Filter when solution is complete. The pH should be approximately 1.6. The solution is usable 10 min after preparation but gives best results in 4-8 wk.
B. Biebrich scarlet WS (CI No. 26905), 0.25 gm; orange G (CI No. 16230), 0.25 gm; phosphotungstic acid, 0.5 gm; phosphomolybdic acid, 0.5 gm; distilled water, 50 ml; 95% ethanol, 50 ml. p H 2.6.
C. Fast green FCF (CI No. 42053), 0.5 gm; 95% ethanol, 100 ml. p H 3.6.
Staining procedure
- Remove embedding material and hydrate to 70% ethanol.
- Stain in solution A for 15-30 min.
- Wash in distilled water.
- Destain brie5y in acid alcohol consisting of: 0.6 N H,S04, 10 ml; distilled water, 50 ml; and 95% ethanol, 40 ml.
- Place for 2 min in a solution consisting of: NaHCO,, 0.5 gm; distilled water, 50 ml; and 95% ethanol, 50 ml.
- Stain in solution B for 15-30 min. The staining time may be adjusted to suit the material.
- Wash in 95% ethanol to remove excess stain.
- Stain in solution C for 15 sec.