GBL® ROSA-STAIN

5021-Mallory-Azan Staining Set

Title Description
REF No 5021
Purpose It is a trichrome staining method used in histological examination of tissues such as connective tissue, cartilage and bone. It is especially preferred to clearly distinguish collagen fibers, muscle fibers and cell nuclei.
Ingredients A) Azocarmine G Solution: Azocarmine G, glacial acetic acid.
B) Aniline-Alcohol Solution: Aniline, Ethanol 96
C) Acetic Alcohol: Glacial acetic acid, Ethanol 100%
D) Phosphotungstic Acid Solution (PTA) Phosphotungstic Acid,
E) Aniline Blue-Orange G Solution Aniline blue, Orange G, Glacial acetic acid
Working Principle The Mallory-Azan Staining Set is based on the principle of selective binding of acidic dyes to tissue components. Different structures within the tissue; nucleus, cytoplasm, collagen, muscle tissue hold different dyes according to their chemical composition (especially protein types and their sensitivity to pH). To obtain quality staining results, it is necessary to stain the section using Azocarmine G Solution and then gradually differentiate using Aniline-Alcohol Solution to ensure that certain structures of the section (such as collagen) are counterstained.
- Azocarmine G Solution: It is an acidic dye that binds to intracellular proteins, mainly nuclei and cytoplasm.
- Phosphotungstic Acid Solution: Removes excess Azocarmine G from tissue, leaving it only in strongly bound areas.
- Aniline Blue: Shows strong affinity for collagen fibers, staining these structures blue.
- Orange G: Stains muscle tissue and erythrocytes, giving orange-yellow tones.
Warnings, Precautions, Recommendations Caution Danger Highly flammable liquid and vapor. Keep away from heat, hot sparks, open flames and other ignition sources. Do not smoke. Causes skin irritation. Causes severe eye irritation. Wear protective gloves/protective clothing/eye protection/face protection. IN CASE OF EYE CONTACT: Rinse thoroughly with water for several minutes. Remove contact lenses if present and easy to remove. Continue rinsing. IF ON SKIN: Wash with plenty of soap and water. If eye irritation persists: Seek medical advice/attention. Keep container tightly closed.
Application 1. Fixation; tissue is fixed with 10% neutral buffered formalin or Bouin's fixative.
2. 4-5 micron thick sections are taken.
3. Deparaffinization and hydration procedures are performed.
4. Rinsed with distilled water for 5 minutes.
5. A-A Azocarmine G Solution is heated to 50-55 °C and the tissue sample is placed in it. Wait for 45-60 minutes.
6. B- Aniline-Alcohol Solution is applied for 1-3 minutes to clarify the nuclei. This is preferably done. This step can be skipped.
7. C- Acetic Alcohol is applied for 1-2 min if step 6 (B- Aniline-Alcohol Solution) will be used. If step 6 is skipped, this step is also skipped.
8. Rinse with distilled water.
9. D) Phosphotungstic Acid Solution (PTA) is applied for 10-15 minutes.
10. Rinse with distilled water.
11. E) Aniline Blue-Orange G Solution is applied for 20-30 minutes.
12. Differentiation can be done with ethanol series. Examined under microscope.
Theoretical Results
Collagen fibers Blue/Green
Muscle fibers Red/Orange
Nucleus Red
Erythrocytes Orange/Yellowish
Stoplasm Pink/Red

Related Media
Shelf Life 2 years
Shelf Life After Opening the Lid
Storage Temperature 15-25 °C
pH (at 25°C)
Transportation Information
Ordering and Packaging Information
Order No Unit Volume Packaging Type Quantity in Box Quantity per Carton
445021050050 Piece 50 ml 50 ml, HDPE White Bottle with Dropper 1 set 20 set

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