HPS, AFIP Modification
General Oversight stain
Also known as the Hematoxylin Phloxine Saffron stain, AFIP modification.
Materials
- Hemalum
- Saturated aqueous picric acid
- Solution A
Material Amount Phloxine B 1.5 g Distilled water 100 mL - Solution B
Material Amount Saffron 2 g Absolute ethanol 100 mL
Tissue Sample
5 µ paraffin sections of neutral buffered formalin fixed tissue are suitable, as are many other fixatives.
Protocol
- Bring sections to water with xylene and ethanol.
- Place in picric acid solution for 5 minutes.
- Wash with water to remove yellow.
- Stain nuclei with hemalum, differentiate and blue.
- Wash well with water.
- Place in solution A for 2 minutes.
- Wash with tap water for 5 minutes.
- Thoroughly dehydrate with absolute ethanol.
- Place into solution B for 5 minutes.
- Rinse well with absolute ethanol.
- Clear in xylene and mount with a synthetic resinous medium.
Expected Results
- Nuclei – blue
- Muscle & cytoplasm – red
- Collagen – yellow
Notes
- The saffron should be extracted by mixing with anhydrous ethanol in a tightly capped container, then placing in a 56°C oven for a few days. Store and use at room temperature. It has a limited life and is best when freshly made.
- Saffron is expensive. It may be available in East Indian grocery stores or health food stores as its most common use today is as a spice and food coloring.
- It is important that there be no water in the saffron solution and that sections be thoroughly dehydrated before it is applied.
- Usually, whole stigmata are more effective than ground saffron.
- Erythrosine B may be used instead of phloxine B.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Histological Staining Methods of the Armed Forces Institute of Pathology, 3rd ed. (1968)
Luna, Lee G.
McGraw-Hill, NY, USA
