Decalcifying Solution, Formic Acid 5%, Aqueous
SOLUTION:
1 Liter | 1 Gallon | 20 Liter Cube | |
Decalcifying Solution, Formic Acid 5%, Aqueous | Part 1049B | Part 1049C | Part 1049E |
Additionally Needed:
Decalcification End Point Set | Part 1051 |
For storage requirements and expiration date refer to individual bottle labels.
APPLICATION:
Newcomer Supply Decalcifying Solution, Formic Acid 5%, Aqueous, provides a moderate rate of decalcification while maintaining cellular morphology. This solution is a general purpose decalcifier and suitable for all bone specimen types from sternal or iliac crest bone marrow biopsies (light bone) to femoral head and long bone sections (compact bone).
METHOD:
Fixation: Formalin 10%, Phosphate Buffered (Part 1090)
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- See Procedure Note #1.
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Technique: Paraffin sections cut at 4 microns on adhesive slides
Solutions: All solutions are manufactured by Newcomer Supply, Inc.
PROCEDURE:
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- Fix bone for a length of time sufficient for specimen size and type.
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- See Procedure Note #2.
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- Adequate bone fixation is essential before decal solution exposure.
- Wash fixed specimen in running tap water for 10 minutes.
- Submerge fixed bone segment in Decalcifying Solution, Formic Acid 5%, Aqueous, covering specimen at a 20:1 ratio.
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- See Procedure Notes #3 and #4.
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- Check specimen regularly for sufficient solution coverage. Change solution daily and do not add or mix fresh solution with old.
- Decalcification time will vary, dependent on bone size and weight.
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- Check light bone samples every 30 to 60 minutes.
- Check compact bone samples every 1 to 2 hours.
- Bone marrow or light bone biopsies, on average, will decalcify in 4 to 6 hours.
- 3 mm thick section of femoral head, on average, will decalcify in 8 to 24 hours.
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- Check decal completion at regular intervals with Decalcification End Point Set (Part 1051) to deter over-decalcification.
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- See Procedure Note #5.
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- Wash in running tap water when decalcification is complete.
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- Wash small samples 30-60 minutes.
- Wash larger bones 1-4 hours.
- Additional trimming of decaled bone can occur at this point to size and thickness suitable for tissue processing.
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- Proceed with tissue processing procedure for bone specimens.
- Trim block and section bone. If trimming or sectioning is impaired due to bone hardness, surface decalcification is recommended.
- Perform surface decalcification: Soak exposed bone surface side down in Decalcifying Solution, Formic Acid 5%, Aqueous for 15-60 minutes. Rinse block with distilled water to remove corrosive acids and re-section.
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- See Procedure Note #6.
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- Fix bone for a length of time sufficient for specimen size and type.
PROCEDURE NOTES:
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- Other fixatives suitable for bone specimens include: AZF Fixative (Part 1009), B-5 Fixative Modified, Zinc Chloride (Part 1015), Bouin Fluid (Part 1020), Zamboni Fixative (Part 1459) and Zinc Formalin Fixative (Part 1482).
- Reduce size of a large bone by bisecting bone into smaller pieces, removing excess soft tissue for faster fixation. Maximum bone thickness of 3-5 mm is recommended.
- Decal solution should be in contact with all specimen surfaces. For multiple pieces, ensure pieces are separated or suspended and not in direct contact or stacked on each other.
- Enhance decal with low-speed agitation shaker, rotator or stir plate.
- Decalcification end-point testing can also be done with specimen radiography. Physical probing of bone is not recommended.
- Only a few calcium-free sections will be obtained after surface decalcification. Repeat the process for additional sections.
REFERENCES:
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- Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 338-343.
- Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 6-11.
- Urban, Ken. “Routine Decalcification of Bone.” Laboratory Medicine 12.4 (1981): 207-212.
- Villanueva, Anthony. “Experimental Studies in Demineralization and Its Effects on Cytology and Staining of Bone Marrow Cells.” The Journal of Histotechnology 9.3 (1986): 155-161.
- Modifications developed by Newcomer Supply Laboratory.