B-5 Fixative Modified, Zinc Chloride
- Shelf Life is 2 years from date of manufacture.
|SOLUTION:||1 Liter||6 X 1 Liter||1 Gallon|
|B-5 Fixative Modified, Zinc Chloride||Part 1015A||Part 1015A||Part 1015C|
|Formaldehyde 37-40%, ACS||Part 1089|
For storage requirements and expiration date refer to individual bottle labels.
Newcomer Supply B-5 Fixative Modified, Zinc Chloride provides an alternative to the classic B-5 fixative with mercuric chloride. Environmental concerns and costly requirements for disposal of mercury-containing fixatives have led to the modification of the original B-5 formulation by substituting zinc chloride for mercuric chloride. This maintains the benefits of B-5 fixation with no concern for mercury pigment.
B-5 Fixative Modified, Zinc Chloride will demonstrate clear nuclear detail, preserve immunohistochemical (IHC) staining and is a good fixative choice for bone marrow, lymph nodes, spleen and other hematopoietic tissues.
Bone Marrow: Recommended fixation time for bone marrow clot is a minimum of 2 hours; for bone marrow biopsy a minimum of 3 hours.
a. See Procedure Note #1.
Lymph Nodes and Small Biopsies: A minimum of 4 hours is recommended. Small nodes (5 mm or less) should be halved. If larger, dissect in such a way that no piece is thicker than 3 mm.
Solutions: All solutions are manufactured by Newcomer Supply, Inc.
- B-5 Fixative Modified, Zinc Chloride Working Solution:
- B-5 Fixative Modified, Zinc Chloride 40 ml
- Formaldehyde 37-40%, ACS 4 ml
- Combine solutions directly before use. Fix tissue in this fresh 10:1 working solution for a minimum of 2 to 4 hours depending upon type of tissue.
- Hold tissue specimens in B-5 Fixative Modified, Zinc Chloride Working Solution until ready to process or for a maximum of 72 hours.
- See Procedure Note #2.
- Wash fixed tissue thoroughly in running tap water to remove residual zinc chloride.
- Place on tissue processor in Formalin 10%, Phosphate Buffered (Part 1090) fixation step.
- Nitric acid solutions are not recommended as a decalcification agent following B-5 Fixative Modified, Zinc Chloride fixation.
- Extended storage of tissue in B-5 Fixative Modified, Zinc Chloride Working Solution is not recommended. After a maximum fixation time of 72 hours, transfer B-5 fixed wet tissue to 70% Ethyl Alcohol (Part 10844) for long-term storage purposes.
- Alternative uses of B-5 Fixative Modified, Zinc Chloride:
a. Stock solution as working solution: Place tissue directly into undiluted B-5 Fixative Modified, Zinc Chloride for 2 to 4 hours. Proceed to Step #3c.
b. Stock solution with Acetic Acid, Glacial, ACS (Part 10010) for working solution: Combine solutions directly before use:
B-5 Fixative Modified, Zinc Chloride 40 ml
Acetic Acid, Glacial, ACS 4 ml
Place tissue in this fresh 10:1 working solution for 2 to 4 hours.
c. Rinse tissue thoroughly in running tap water. Proceed with Formalin 10%, Phosphate Buffered (Part 1090) fixation on tissue processor.
d. Without the addition of formaldehyde to B-5 Fixative Modified, Zinc Chloride, the quality of tissue fixation may be compromised.
- Neutralize B-5 Fixative Modified, Zinc Chloride, stock or working solutions with magnesium hydroxide/oxide, sodium carbonate or sodium bicarbonate to precipitate zinc at pH 7.0-8.0. Separate solids from liquid and dispose of according to local and state environmental regulations.
- Approximately 100 grams of sodium bicarbonate will neutralize/precipitate zinc from 1 liter of B-5 Fixative Modified, Zinc Chloride.
- Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 69.
- Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 17-19.
- Dapson, Janet Crookham, and Richard Dapson. Hazardous Materials in the Histopathology Laboratory: Regulations, Risks, Handling, and Disposal. 4th ed. Battle Creek, MI: Anatech, 2005. 148-149, 279.
- Modifications developed by Newcomer Supply Laboratory.