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.
Nuclei – blue
Muscle & cytoplasm – red
Collagen – yellow
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 colouring.
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.
Reference Histological Staining Methods of the Armed Forces Institute of Pathology, 3rd ed. (1968) Luna, Lee G.
McGraw-Hill, NY, USA