URMC Pathology/Morphology Imaging Core (PMIC)

MOWIOL

Recommended Anti-Fade for Fluorescent Microscopy, especially EGFP

Mowiol goes into solution with difficulty.  It’s best to make a large batch and freeze aliquots at -20·C.

Prepare in clean 250 ml flask or beaker

1) 24 g analytical grade glycerol (Sigma #G-6279)

2)   9.6 g Mowiol 4-88 (Fluka, #81381 (through Sigma))

3) 24 ml distilled water

4) 48 ml 0.2M Tris buffer, pH 8.5

5) 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.

6) Aliquot into 50 ml centrifuge tubes, weigh  and balance

7) Centrifuge at 5000g for 15 min., there will be a pellet at the base, carefully remove the supernatant

8) Aliquot into 15 ml conical tubes – add only 10 mls for expansion.

9) Store at -20 for 12 months. Store at room temperature no more than one month

10) Use Mowiol that has been warmed to room temperature to eliminate bubble formation. For an 18 mm coverslip, use approximately 10 ul Mowiol (small amount only). Dry slides flat overnight at room temperature, well covered.

Note:  There are a number of recipes available for making Mowiol in 50 ml conical tubes – but we have found this method a problem for getting the Mowiol into solution.  Therefore the recipe was changed to the above 200ml volume and our results for getting Mowiol to go into solution have substantially improved. 

URMC In-House Sources:  Michael Mastrangelo, Eric Yehling