- Catalog order # K7184 - 4oz kit - $56.00
- K7188 - 8oz kit - $97.00
APPLICATION
Mammalian Tissue
DESIGNED TO SHOW
Ionic Iron Deposits
SPECIMEN REQUIREMENTS
Fixation: Tissue: Any well fixed tissue, however, it is suggested to use formalin fixed tissue.
Sections: Cut Paraffin sections at 5 microns.
COMPONENTS AND REAGENTS
1. Stock Hydrochloric Acid Solution - 4 oz / 120 ml
2. Stock Potassium Ferrocyanide - 4 oz / 120 ml
3. Nuclear Fast Red - 4 oz / 120 ml
* Iron Solution (Working)
Combine 25ml of Stock Potassium Ferrocyanide Solution with 25 ml of Stock Hydrochloric Acid Solution. Mix.
Prepare solution just before use.
STAINING PROCEDURE
1. Deparaffinize sections and hydrate to deionized water.
2. Treat section in *Iron Solution (Working) for 30 minutes.
3. Rinse section thoroughly through 6 changes of distilled water.
4. Counterstain section in Nuclear Fast Red for 4 minutes.
5. Rinse sections in deionized water for at least 3-4 minutes.
6. Dehydrate sections through alcohols.
7. Clear section in Xylene or Xylene replacement.
(Xylene alternatives require increased time schedules)
8. Mount with Clearslip.
RESULTS
Ferric salts and Hemosiderin Blue
Nuclei, erythrocytes, connective tissue, cytoplasm-Shades of pink
NOTES
1. Bone decalcifiers will have an impact on staining results. Do not allow bone tissue to soak in Decalcifying solutions for prolonged periods of time. Reduce the size of bone width to decrease decalcification times. Microwaving completely fixed tissue sample using decalcification solutions as Formic/Hydrochloric Decalcifiers can reduce decalcification time to 10 minutes. Fill beaker with decalcification solution and treat bone tissue (Enough to fill a routine cassette). Microwave on Medium to High, pending on type of microwave system, two five minute intervals allowing tissue to soak in heated solution for 10 minutes between each change.
REFERENCES
1. Gomori, G (1936): Am J. Path.; 12:pp. 655-663
2. Perls, M.V. (1967): Virchow,s Arch. Path. Anat; 39:42
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