Sudan Black B Stain, Propylene Glycol
(use: Stains Neutral Lipids & Phospholipids.)
SOLUTION:
125 ml | 250 ml | |
Sudan Black B Stain, Propylene Glycol | Part 1401A | Part 1401B |
Additionally Needed:
Formalin 10%, Phosphate Buffered | Part 1090 |
Propylene Glycol 100%, ACS | Part 13391 |
Propylene Glycol 85%, Aqueous | Part 133912 |
Nuclear Fast Red Stain, Kernechtrot | Part 1255 |
Mount-Quick Aqueous | Part 6271A |
For storage requirements and expiration date refer to individual bottle labels.
APPLICATION:
Newcomer Supply Sudan Black B Stain, Propylene Glycol procedure is used for identification of fat/lipid in frozen sections. Sudan black B is a lipid soluble solvent dye that readily stains neutral fats and phospholipids.
Sudan dyes are a group of fat/lipid soluble solvent dyes, also known as lysochromes. These solvent dyes readily stain fat/lipid since the dyes are more soluble in lipid than in the solvents from which they are applied.
METHOD:
Fixation: Fresh tissue or formalin fixed unprocessed tissue
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- See Procedure Notes #1and #2.
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Technique: Frozen sections cut at 8-10 microns on adhesive slides
Solutions: All solutions are manufactured by Newcomer Supply, Inc.
All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.
STAINING PROCEDURE:
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- Fix frozen section slides in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
- Rinse sections in two changes of distilled water.
- Blot off excess water and dehydrate slides in Propylene Glycol 100%, ACS (Part 13391) for 10-15 minutes.
- Place directly into Sudan Black B Stain, Propylene Glycol for 30 minutes to 1 hour. Agitate occasionally or place Coplin jar on rotator/shaker with continuous gentle agitation.
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- See Procedure Note #3.
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- Differentiate in Propylene Glycol 85%, Aqueous (Part 133912) for 3 minutes with agitation.
- Rinse in distilled water.
- Counterstain in Nuclear Fast Red Stain, Kernechtrot (Part 1255) for 5 minutes.
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- Shake solution well before use; do not filter.
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- Wash gently in several changes of tap water.
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- See Procedure Note #4.
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- Blot excess water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) mounting medium.
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- See Procedure Notes #5 and #6.
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RESULTS:
Fat | Blue-black |
Nuclei | Red |
PROCEDURE NOTES:
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- Freeze tissue according to laboratory protocol.
- To freeze formalin fixed tissue that has not been processed:
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- Wash tissue in running tap water for 5 minutes.
- Blot excess water from tissue and freeze.
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- To decrease staining time; preheat Sudan Black B Stain, Propylene Glycol in a 60°C oven; stain for 3-10 minutes.
- Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in sections.
- Use minimal pressure with coverslip application to avoid displacement of fat/lipid staining.
- To remove trapped air bubbles or to recoverslip;
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- Soak slide in warm water until coverslip falls off.
- Blot excess water from slide.
- Remount with new coverslip and Mount-Quick Aqueous.
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REFERENCES:
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- Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 192-193.
- Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 185-186.
- Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 464-465.
- Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 204-205.
- Modifications developed by Newcomer Supply Laboratory.