B-5 Fixative Modified, Zinc Chloride

 

  • Shelf Life is 2 years from date of manufacture.

 

Product Options:

1 - 5:
Part # 1015A 1 L $48.80
6 or more:
Part # 1015A 1 L $42.80
Each:
Part # 1015C 1 gal $97.40

 

SOLUTION: 1 Liter 6 X 1 Liter 1 Gallon
B-5 Fixative Modified, Zinc Chloride Part 1015A Part 1015A Part 1015C

 

Additionally Needed:

Formaldehyde 37-40%, ACS Part 1089

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

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.

 

METHOD:

Fixation:

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.

 

FIXATION PROCEDURE:

  1. B-5 Fixative Modified, Zinc Chloride Working Solution:
  1. B-5 Fixative Modified, Zinc Chloride            40 ml
  2. Formaldehyde 37-40%, ACS                            4 ml
  1. 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.
  2. Hold tissue specimens in B-5 Fixative Modified, Zinc Chloride Working Solution until ready to process or for a maximum of 72 hours.
  1. See Procedure Note #2.
  1. Wash fixed tissue thoroughly in running tap water to remove residual zinc chloride.
  2. Place on tissue processor in Formalin 10%, Phosphate Buffered (Part 1090) fixation step.

PROCEDURE NOTES:

  1. Nitric acid solutions are not recommended as a decalcification agent following B-5 Fixative Modified, Zinc Chloride fixation.
  2. 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.
  3. 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.

  1. 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.
  1. Approximately 100 grams of sodium bicarbonate will neutralize/precipitate zinc from 1 liter of B-5 Fixative Modified, Zinc Chloride.

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 69.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 17-19.
  3. 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.
  4. Modifications developed by Newcomer Supply Laboratory.