Canaliculi and Lacunae Stain for Hard Tissues

EMS26201

Components:

Cupric Nitrate 1% Aqueous

Protargol Solution 1% Aqueous

Copper Shot

Hydroquinone Reducing Solution

Gold Chloride 1% Aqueous

Oxalic Acid 2% Aqueous

Sodium Thiosulfate 5% Aqueous

Fixation:

Modified Bouin's Solution

Decalcification:

Trichloroacetic Acid – Alcohol

Embedded in:

Paraffin, ester wax, celloidin or gelatin

Procedure:

1. Deparaffinise and hydrate to distilled water.
2. Mordant in Cupric Nitrate, 1% overnight.
3. In a coplin jar, add 4-6 gm Copper Shot per 100ml of Protargol Solution and place slides in the solution. Place in a 60°C oven. Staining is complete when the sections have a golden brown color (4-16 hours). Wash very briefly in distilled water (1 dip).
4. Reduce in Hydroquinone Solution for 3-10 minutes. Wash thoroughly in running water to remove all of the reducing solution.
5. Tone 5-10 minutes in Gold Chloride, 1% to which 3 drops of Glacial Acetic Acid per 100 ml have been added. Rinse in water.
6. Develop in Oxalic Acid, 2% until sections have a faint purple or blue color. Rinse in water.
7. Place in Sodium Thiosulfate, 5% for 5-10 minutes. Wash thoroughly in water.
8. Dehydrate, clear and cover.

Stain Results:

References:

Powers, M.M., Rasmussen, G.L., and Clark, G., Anat.Rec., 111:171-6, 1951
Clark, G. (ed), Staining Procedures, 3rd edition, Williams & Wilkins, Baltimore, c 1973, p 136
Bodian, D., Anat Rec., 65:89-97, 1936
Bodian, D., Anat. Rec., 69:153-62, 1937