ST. ALEXIUS COLLEGE, INC.

ACTIVITY NO. 3
DECALCIFICATION

I. INTRODUCTION:
Cutting of thin sections by ordinary method is impossible when the tissue has become partly
calcified. Decalcification is the procedure whereby calcium or lime salts are removed from tissues (most
especially bones and teeth) following fixation. This is usually carried out by the use of chemical agents,
either with acids to form soluble calcium salts, or with chelating agents that bind to calcium ions.
Decalcification should be done after fixation and before impregnation, to ensure and facilitate the
normal cutting of sections and to prevent obscuring the microanatomic detail of such sections by bone
dust and other cellular debris.
Methods of decalcification involve the use of acids, in which the bone salts are dissolved. All
such acid solutions are injurious to the organic ground substances of the tissues, which must therefore be
protected by adequate fixation before decalcification.
A good decalcifying agent must be capable of removing calcium salts from tissues completely
without producing considerable destruction of cells and tissue components and without adversely
affecting the staining capacity of the cell, particularly of the nucleus.

II. OBJECTIVES:
At the end of this laboratory exercise, the student is expected to:
1) discuss the importance of decalcification;
2) determine the various chemicals used for decalcification;
3) perform decalcification appropriately; and
4) troubleshoot decalcification problems.

III. MATERIALS:
Dehydrating agent
Tissue specimen
Timer

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IV. PROCEDURE:

1. Wear a laboratory gown.

2. Wash hands, wear mask, gloves, head cap, goggles and apron.

3. Remove the tissue from fixative and rinse in tap water of PBS.

4. Place the tissue in a 10% formic acid solution until tissue is determined to be decalcified.

5. Use a large volume of solution to tissue. Change the solution daily if the decalcification process
takes more than one day. This could take anywhere from a few hours to several days.

6. Remove tissue from formic acid and rinse in running tap water for at least 10 minutes.

7. Place the tissue in 70% ethanol for storage and delivery.

8. Dispose used and contaminated blood collection materials on appropriate waste container.

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REPORT SHEET
ACTIVITY NO. 3
DECALCIFICATION

NAME: __________________________________________ RATING: ___________

DATE PERFORMED: __________________ DATE SUBMITTED: ______________

ILLUSTRATIONS:

Draw and label the decalcification procedure.

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REVIEW QUESTIONS:

1. Enumerate at least three (3) advantages and disadvantages of at least two decalcifying agents.

DECALCIFYING
ADVANTAGES DISADVANTAGES
AGENTS

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2. What happens during post decalcification?

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3. What is tissue softener? Explain its importance.

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4. What are the usual problems during decalcification? Discuss how to troubleshoot each.

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ST. ALEXIUS COLLEGE, INC.

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 ST. ALEXIUS COLLEGE, INC.

ACTIVITY NO. 3
DECALCIFICATION
Performance Assessment Sheet

Name:_________________________________________ Date:_________________________

PROCEDURE
You must: RATING COMMENTS
1. Wear a laboratory gown.
2. Wash hands, wear mask, gloves, head cap,
goggles and apron.
3. Remove the tissue from fixative and rinse in
tap water of PBS.
4. Place the tissue in a 10% formic acid solution
until tissue is determined to be decalcified.
5. Use a large volume of solution to tissue.
Change the solution daily if the decalcification
process takes more than one day. This could
take anywhere from a few hours to several
days.
6. Remove tissue from formic acid and rinse in
running tap water for at least 10 minutes.
7. Place the tissue in 70% ethanol for storage and
delivery.
8. Dispose used and contaminated blood
collection materials on appropriate waste
container.
RATING:

4 – EXCELLENT
3 – VERY SATISFACTORY
2 – SATISFACTORY
1 – POOR
0 – NOT DONE

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LEVEL OF COMPETENCY:

Evaluator: Evaluation Date:

Student:

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