Figure 1. Equine cutaneous amyloidosis, Congo red, non-polarized left, polarized right.
Brown – Hopps (Gram) Staining Procedure for Bacteria
Purpose:
This standard operating procedure (SOP) establishes the standard for staining
amyloid in using the Bennhold’s Congo Red staining procedure.
Reagents and Materials:
Reagents
1%Congo Red
Congo Red 1.0gm
Distilled water 100ml
Saturated Lithium Carbonate
Lithium Carbonate 1.3gm
Distilled water 100ml
Hematoxylin
Commercially prepared by Anatech Co.
Expiration date and lot number on bottle.
Store at room temperature.
Materials
Tissue slides and control slides for special stain requested.
Microwave to bake slides.
Staining rack, 20 slide capacity.
Coverslips and synthetic mounting media.
Hazards
and Precautions:
See Material Safety Data Sheets.
Sample
Preparations:
Paraffin tissue section slides cut at 4 microns.
Calibration
and Standardization:
A tissue section slide containing amyloid is stained with each procedure.
Procedure:
1. Deparaffinize
and hydrate to distilled water.
2. Congo Red - 30minutes.
3. Saturated Lithium Carbonate - 10 seconds.
4. Tap water rinse
5. Hematoxylin - 1 minute.
6. Proceed as routine staining omitting Eosin.
7. Distilled,clear and coverslip.
Results:
Amyloid - pale pink to red.
Nuclei - blue
Figure
1. Equine cutaneous amyloidosis, Congo red, non-polarized left, polarized
right.
Effective: July
1, 2004
Cherie Chapman
James R. Turk
References:
Manual of Histologic and Special Staining Technics, New York, The Blakiston Division, McGraw- Hill Book Company, Inc., 1960, page 146.Lillie, R.D. Histopathologic Technic, and Practical Histochemistry, New York, The Blakiston Company, Inc., 1954, page 293.