Sudan Black B Stain, Propylene Glycol
(use: Stains Neutral Lipids & Phospholipids.)
|SOLUTIONS:||125 ml||250 ml|
|Sudan Black B Stain, Propylene Glycol||Part 1401A||Part 1401B|
|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 Mounting Medium||Part 6271A|
For storage requirements and expiration date refer to individual bottle labels.
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 due to the fact that the dyes are more soluble in lipid than in the solvents from which they are applied.
Fixation: Fresh tissue or formalin fixed unprocessed tissue
- See Procedure Note #1.
â€‹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.
- Fix frozen section slides in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
- See Procedure Note #2.
- Rinse sections carefully 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.
- See Procedure Note #3.
- Differentiate in Propylene Glycol 85%, Aqueous (Part 133912) for 3 minutes with agitation.
- Rinse gently in distilled water.
- Counterstain in Nuclear Fast Red Stain, Kernechtrot (Part 1255) for 5 minutes.
- Shake solution well before use; do not filter.
- Wash gently in several changes of tap water.
- See Procedure Note #4.
- Blot excess water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) Mounting Medium.
- See Procedure Note #5.
- To freeze formalin fixed unprocessed tissue:
- Place specimen in tissue cassette, wash in running tap water for 5 minutes.
- Remove tissue from cassette; blot well, removing all excess water from tissue.
- Freeze tissue according to laboratory protocol.
- Frozen formalin fixed tissue does not require additional formalin fixation.
- To decrease staining time; preheat Sudan Black B Stain, Propylene Glycol in a 60oC oven; stain for 3-10 minutes.
- Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in coverslipped sections.
- Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed. To remove trapped air bubbles or to recoverslip;
- Soak slide(s) in warm water until coverslip is easily removed.
- Blot excess water from slide.
- Remount with new coverslip and Mount-Quick Aqueous Mounting Medium.
- 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.