Ethylenediaminetetraacetic acid in ammonium hydroxide for reducing decalcification time.
Ethylenediaminetetraacetic acid (EDTA) solution is used to decalcify bone specimens for histological examination. Sodium hydroxide (NaOH) has been used to dissolve EDTA and to bring EDTA solutions to neutral pH. This solution, however, requires several weeks to decalcify bone specimens. We investigated a new decalcification fluid using concentrated ammonium hydroxide (NH4OH) to dissolve EDTA and to adjust the pH to neutral. Decalcification was performed using a magnetic stirrer with and without vacuum, or with a sonic cleaner. Decalcification end point was confirmed using both the weight loss and X-ray methods. After decalcification, specimens were processed through paraffin and sections were stained with hematoxylin and eosin. Decalcification employing NH4OH required an average of six days. Light microscopy indicated good retention of cellular detail.
Sanderson
C
C
Bone and Joint Research Laboratory, Veteran Affairs Medical Center, Salt Lake City, Utah 84148, USA.
Radley
K
K
Mayton
L
L
D064753:Ammonium Hydroxide; D001842:Bone and Bones / Q000737:chemistry* / Q000166:cytology; D002356:Cartilage / Q000737:chemistry* / Q000166:cytology; D003648:Decalcification Technique; D004121:Dimethyl Sulfoxide; D004492:Edetic Acid; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006878:Hydroxides; D013194:Staining and Labeling
D006878:Hydroxides; D064753:Ammonium Hydroxide; D004492:Edetic Acid; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D004121:Dimethyl Sulfoxide