
APPLICATION
Mammalian Tissue
DESIGNED TO SHOW
Detection of Amyloidosis
SPECIMEN REQUIREMENTS
Fixation: Tissue: Any well fixed tissue , however, it is suggested to use formalin fixed tissue.
Sections: Cut Paraffin sections at 5 microns. Thicker sections illustrate brighter staining and minute amyloid deposits
COMPONENTS AND REAGENTS
1. Nuclear One Hematoxylin - 4oz / 120ml
2. Congo Red Stain Solution - 4oz / 120ml
3. Alcohol Solution - 4oz / 120ml
4. Stock Sodium Hydroxide - 4oz / 120ml
5. 1 Stained and 3 unstained control slides
6. Pap Jars
* Alkaline Alcohol Solution
Add 0.5 ml of Stock Sodium Hydroxide to every 60ml of Alcohol Solution.
STAINING PROCEDURE
1. Deparaffinize sections and hydrate to deionized water.
2. Stain sections in Congo Red Stain Solution for 1 hour.
3. Rinse off excess stain in deionized water.
4. *Differentiate sections in * Alkaline Alcohol Solution by dipping 2-3 times with agitation. Background should appear clear.
5. Wash in deionized water for 5 minutes with agitation.
6. Counterstain in Nuclear One Hematoxylin for 5 minutes.
7. Wash excess stain and blue sections in deionized water for 5 minutes.
8. Dehydrate sections in one change of 95% Reagent Alcohol and three changes of Absolute Alcohol.
9. Clear sections through two changes of Xylene.
10. Mount using Clearslip.
RESULTS
Amyloid - Red to pink
Nuclei- Blue
NOTES
1. *Alkaline Alcohol Solution should be made fresh before use. Solution should be discarded after each use.
REFERENCES
1. AFIP, Manual of Histologic Staining Methods, McGraw Hill Book Co., 1968; pp. 168-169
Home Page