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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
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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.