Sudan IV Stain, Herxheimer Alcoholic

Product Options:

Part # 1400A 125 ml $31.80 H

SOLUTIONS: 125 ml
Sudan IV Stain, Herxheimer Alcoholic Part 1400A

 

Additionally Needed:

Alcohol, Ethyl Denatured, 70% Part 10844
Hematoxylin Stain, Mayer Modified Part 1202
Lithium Carbonate, Saturated Aqueous

                 OR

Scott Tap Water Substitute

Part 12215

    OR

Part 1380

Mount-Quick Aqueous Mounting Medium Part 6271A
For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Sudan IV Stain, Herxheimer Alcoholic procedure is used for identification of fat/lipid in frozen sections. Herxheimer method refers to an acetone/alcohol solvent mixture; the acetone component of this solution may dissolve out small amounts of lipid.

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.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue

  1. See Procedure Note #1.

​Technique: Frozen sections cut at 8 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:

  1. Fix frozen section slides in Formalin 10%, Phosphate Buffered for 1 minute.
    1. See Procedure Note #2.
  2. Rinse sections carefully in two changes of distilled water.
  3. Rinse in Alcohol, Ethyl Denatured, 70% (Part 10844).
  4. Stain in Sudan IV Stain, Herxheimer Alcoholic for 10 minutes.
    1. Keep tightly capped to avoid evaporation.
  5. Differentiate quickly in Alcohol, Ethyl Denatured, 70% to remove excess stain.
  6. Wash thoroughly in distilled water.
  7. Counterstain with Hematoxylin Stain, Mayer Modified (Part 1202) for 2-3 minutes.
  8. Wash gently in several changes of tap water.
  9. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
    1. The use of a bluing agent is optional.
  10. Wash gently in several changes of tap water.
  11. Blot excess water from slide; coverslip with Mount-Quick Aqueous Mounting Medium.
    1. See Procedure Note #3.

 

RESULTS:

Fat Orange/red
Nuclei Blue

 

PROCEDURE NOTES:

  1. To freeze formalin fixed unprocessed tissue:
    1. Place specimen in tissue cassette; wash in running water for 5 minutes.
    2. Remove tissue from cassette; blot well, removing all excess water from tissue.
    3. Freeze tissue according to laboratory protocol.
  2. Frozen formalin fixed tissue does not require additional formalin fixation.
  3. Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed.  To remove trapped air bubbles or to recoverslip;
    1. Soak slide in warm water until coverslip is easily  removed.
    2.  Blot excess water from slide.
    3. Remount with new coverslip and Mount-Quick Aqueous Mounting Medium.

 

REFERENCES:

  1. Culling, C. F. A. Handbook of Histopathological and Histochemical Techniques: (including Museum Techniques). 3rd ed. London: Butterworth, 1974. 359-362.
  2. Kiernan, J. A. Histological and Histochemical Methods: Theory and Practice. 3rd ed. London, Ontario: Arnold, 2003. 251-254.
  3. Lillie, R. D., and Harold Fullmer. Histopathologic Technic and Practical Histochemistry. 4th ed. New York: McGraw-Hill, 1976. 565-568.
  4. Modifications developed by Newcomer Supply Laboratory.