Products

Oil Red O Stain, Propylene Glycol

Newcomer Supply Beaker

(use: Stains neutral lipids.)

Product Options:

Part # 12772A 250 ml $48.20
Part # 12772B 500 ml $84.60

SOLUTION:

250 ml 500 ml
Oil Red O Stain, Propylene Glycol Part 12772A Part 12772B

 

Additionally Needed:

Formalin 10%, Phosphate Buffered Part 1090
Propylene Glycol 100%, ACS Part 13391
Propylene Glycol 85%, Aqueous Part 133912
Hematoxylin Stain, Mayer Modified Part 1202
Lithium Carbonate, Saturated Aqueous
               OR
Scott Tap Water Substitute		 Part 12215
    OR
Part 1380
Mount-Quick Aqueous Part 6271A

     

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Oil Red O Stain, Propylene Glycol procedure is classified as a physical staining method. Oil Red O (ORO) staining is used for identification of fat in frozen tissue sections.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue
Technique: Frozen sections cut at 8-10 microns on adhesive slides

        1. See Procedure Notes #1 and #2.

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.

 

PRESTAINING PREPARATION:

    1. Preheat Oil Red O Stain, Propylene Glycol in 60oC oven for a minimum of 30 minutes. Save for Step #7.
    2. If a film or precipitate forms on Oil Red O Stain, Propylene Glycol, skim the surface with filter paper before use.

 

STAINING PROCEDURE:

    1. Fix frozen section in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
    2. Rinse sections in two changes of distilled water.
    3. Blot water and place in Propylene Glycol 100%, ACS (Part 13391) for 2-5 minutes with agitation.
    4. Room Temperature ORO Staining: Place in Oil Red O Stain, Propylene Glycol for 1 hour with agitation.
    5. Heated ORO Staining: Place in preheated Oil Red O Stain, Propylene Glycol (Step #1) for 7-10 minutes. Agitate occasionally.
    6. Differentiate in Propylene Glycol 85%, Aqueous (Part 133912) for 3 minutes with agitation.
    7. Rinse in two changes of distilled water.
    8. Stain in Hematoxylin Stain, Mayer Modified (Part 1202); 2-3 minutes.
    9. Wash in several changes of tap water.
    10. Optional: Blue in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
    11. Wash in several changes of tap water.
    12. Blot water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) mounting medium.
        1. See Procedure Notes #3 and #4.

 

RESULTS:

Fat Bright red
Nuclei Blue to dark blue

 

PROCEDURE NOTES:

    1. Freeze tissue according to laboratory protocol.
    2. To freeze formalin fixed tissue that has not been processed:
        1. Wash tissue in running tap water for 5 minutes.
        2. Blot excess water from tissue and freeze.
    3. Use minimal pressure when applying coverslip to avoid displacement of fat/lipid staining.
    4. To remove trapped air bubbles or to recoverslip;
        1. Soak slide in warm water until coverslip falls off.
        2. Blot excess water from slide.
        3. Remount with new coverslip and Mount-Quick Aqueous.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 184-186.
    2. Prophet, Edna B., Bob Mills, Jacquelyn Arrington and Leslie Sobin. Laboratory Methods in Histotechnology. Washington, D.C.: American Registry of Pathology. 1992.178.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 205.
    4. Modifications developed by Newcomer Supply Laboratory.