• Spread tissue on glass slide
• Fix in Carnoy's fixative for 2 to 4 hours at r.t.
• Wash in 70 % EtOH for 15 min
• Change gradually to distilled water
• Rinse in distilled water for 5 min
• Stain in carmine alum O/N
• Wash in 70 % EtOH 15 min
• Wash in 95 % EtOH 15 min
• Wash in 100 % EtOH 15 min
• Clear in xylene and mount with Permount

After photographic documentation the tissue can be embedded in paraffin for sectioning and conventional histological staining.

• Immerse slide into xylene to remove mounting medium
• Change xylene 2 times
• Transfer to 1:1 xylene:paraffin (60°C)
• Change xylene 2 times and embed in paraffin

Carnoy's Fix:
6 parts 100 % EtOH
3 parts CHCl3
1 part glacial acetic acid

Carmine Alum Stain:
Place 1 g carmine (Sigma C1022) and 2.5 g aluminum potassium sulfate (Sigma A7167) in 500 ml distilled water and boil for 20 min. Adjust final volume to 500 ml with water. Filter and add a crystal of thymol as preservative. Refrigerate. Can be used for several months. Discard when color becomes weak.