TISSUE DECALCIFICATION.
DRAFT COPY ONLY.
(Procedure 20).
http://www.netautopsy.org/axsop/axsop020.htm


NEXT PAGE
PREVIOUS PAGE
RETURN TO TABLE OF CONTENTS

United States Government Work, uncopyrighted, public-domain, DRAFT COPY ONLY. This document does not necessarily represent the views or policies of any United States Government agency. This document is provided "as is", without warranty of any kind, express or implied, including but not limited to the warranties of merchantability, fitness for a particular purpose and non-infringement. In no event shall the authors be liable for any claim, damages or other liability, whether in an action of contract, tort or otherwise, arising from, out of, or in connection with the document or the use or other dealings made with the document.

See also: Main objectives: 1.



PRINCIPLE OF THE TEST.


Preparation of calcified tissue for histology on bone and calcified tissue specimens.



SPECIMEN REQUIRED.


All human tissue excised at surgery, outpatient clinics, and postmortems, fresh or in fixative, along with a filled-out Tissue Examination Form (U. S. Standard Form 515, USSF515).



REAGENTS, INSTRUMENTATION.


Decalcifying solution (RDO).



STEP-BY-STEP DESCRIPTION.


1. Submit only what is necessary for decalcification! If possible, large specimens should be trimmed to appropriate size, prior to fixation and decalcification. Specimens should be allowed to fix overnight, then washed in running water. Completely immerse the specimen in 50 - 150 cc. of RDO solution in a plastic or glass container. Changing of solution is not required; however, decalcification can be facilitated by occasional agitation. Cancellous and thin cortical bone should be decalcified in 4 hours or less (larger specimens will, of course, require more time as will thick, dense specimens). After decalcification is complete, thoroughly wash the specimen for 2 - 3 hours in running water. Submit for routine processing.

2. Do NOT overdecalcify!!! Specimens should not remain in RDO for more than 12 hours. Do not leave specimen in RDO overnight.

3. Complete decalcification can be determined by testing with a sharp needle, or by cutting with a blade. The absence of gritty resistance indicates that all mineral has been dissolved.



REFERENCE.


      1. Prophet EB, Mills B, Arrington JB, Sobin LH.
Laboratory Methods in Histotechnology, pp. 71-79.
1992: Armed Forces Institute of Pathology, Washington, DC. 20306-6000.
ISBN: 1-881041-00-X 1992.