A semi-permanent mounting media for immunofluorescence microscopy.

Method

1. Add the following reagents to a 250 ml flask or beaker:

• 24 g analytical grade glycerol (Sigma #G-6279)
• 9.6 g Mowiol 4-88 (Fluka, #81381, can be purchased through Sigma-Aldrich)
• 24 ml distilled water
• 48 ml 0.2M Tris buffer, pH 8.5

2. Stir with a clean stir bar on a hot plate on warm (not boiling)at least 4-5 hours until the majority of the Mowiol powder goes into solution.
3. Aliquot into 50 ml centrifuge tubes, weigh and balance
4. Centrifuge at 5000g for 15 minutes. Carefully remove the supernatant without disturbing the pellet at the bottom of the flask.
5. Aliquot into 15 ml conical tubes – add only 10 mls to each tube to allow for expansion with freezing.
6. Aliquots may be stored at -20C for 12 months. Store at room temperature no more than one month.
7. To use, warm solution to room temperature to eliminate bubble formation. use approximately 10 ul of mounting media for an 18 mm coverslip.
8. Allow slides to dry overnight at room temperature in a light-tight box.