Crystal Violet Stain, Lieb, Alcoholic
(use: Easy, backup Amyloid Stain.)
SOLUTION:
100 ml | |
Crystal Violet Stain, Lieb, Alcoholic | Part 10421A |
Additionally Needed:
Amyloid, Animal Control Slides | Part 4031 |
Xylene, ACS | Part 1445 |
Alcohol, Ethyl Denatured, 100% | Part 10841 |
Alcohol, Ethyl Denatured, 95% | Part 10842 |
For storage requirements and expiration date refer to individual product labels.
APPLICATION:
Newcomer Supply Crystal Violet Stain, Lieb, Alcoholic is used to provide a rapid screening method for amyloid deposits in tissue sections. This procedure has low sensitivity and should be considered as an amyloid screening technique and not an amyloid specific stain.
METHOD:
Fixation: Formalin 10%, Phosphate Buffered (Part 1090)
Technique: Paraffin sections cut at 10-12 microns
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- See Procedure Note #1.
Solutions: All solutions are manufactured by Newcomer Supply, Inc.
STAINING PROCEDURE:
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- If necessary, heat dry tissue sections/slides in oven.
- Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.
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- See Procedure Note #2.
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- Stain sections in Crystal Violet Stain, Lieb, Alcoholic for 10 minutes.
- Rinse well in tap water.
- Blot water from slides; allow slides to air-dry in a vertical position.
- Coverslip air-dried sections with compatible mounting medium.
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- See Procedure Note #3.
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RESULTS:
Amyloid | Purple/violet |
Background | Purple/blue |
PROCEDURE NOTES:
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- For best results cut sections at 10-12 microns to provide better definition and more intense amyloid staining.
- Drain slides after each step to prevent solution carry over.
- Avoid the use of aqueous based mounting mediums which will cause bleeding/diffusion of the stain from the tissue section.
- If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization step.
REFERENCES:
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- Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 154-155.
- Lieb, E. “Permanent Stain for Amyloid.” American Journal of Clinical Pathology 17 (1947). 413-414.
- Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 154.
- Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 177.
- Modifications developed by Newcomer Supply Laboratory.