[Histonet] Eosin stain for determining new bone old bone

Jacquitta Taylor jacquitta <@t> earthlink.net
Wed Feb 2 09:13:40 CST 2011

Robin this is the protocol we use to showing new bone old bone.

Procedure for Eosin Y 0.6 % For Staining Bone Specimen
EosinY, Sigma E-4382,
100% Ethanol,
Phloxine B (Sigma P-4030)
Orange G(sodium salt-Sigma O-1625)
Stock 0.6% Eosin
Add 6g of eosin to 900ml 100% Ethanol and100 ml of H20; stir to dissolve.
Add 50 ml glacial acetic acid adjust PH to 4.6 and 5.0. The color of the solution will change from opaque green to clear red.
Stock 1% Phloxine B Solution:
Dissolve 1g of Phloxine B in 100 ml distilled water and filter to make a 1% solution.
Stock 2% Orange G Solution:
Dissolve 2 g of Orange G in 100ml distilled water and filter to make a 2% solution.
Working Solution Eosin 0.6%
Make a working solution by adding 6 ml of 1% Phloxine and 6 ml of 2% Orange G to 238 ml of 0.6% eosin.
Old bone dark orange.
New bone light orange.
Stain 15 to 30 seconds depending on what intensity you prefer.

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