[Histonet] Optimizing H&E Stains for undecalcified bone

[M.O.]

Hello Histonetters!  I am new to staining and can't seem to get good H&E
stains from deplacticized undecalcified bone with an implant.  It seems as
though the hematoxylin is staining too dark. Any feedback would be much
appreciated!

The protocol I am using is for after the deplasticization step and says:

100% EtOH for 3 mins, 70% EtOH for 3 mins

Hematoxylin for 6mins, Rinse, .25% Acid Ethanol dip 8-10 times, Rinse

95% EtOH 1min

Eosin for 2mins

Quick Dip 95%, 95%, 100% 100% EtOH

Soak in ProPar for 15+mins


Sincerely,
Merissa