Aqueous mounting medium for fluorescence and Stereology. Recommended Anti-Fade for fluorescence microscopy – especially good for EGFP and other low expression reporting molecules.
Mowiol goes into solution with difficulty. It’s best to make a large batch and freeze aliquots at -20ºC.
Takes about 8 hours to prepare correctly.
Note for stereology: if you have sections mounted in DPX – remove the DPX by soaking and washing in zylene, rehydrate in PBS and Azide (for 1-2 nights) then mount in Mowiol
Add 24g glycerol (analytical grade) to clean 250 ml flask or beaker
Add 9.6g Mowiol 4-88 (Fluka, #81381 – through Sigma) and stir thoroughly to mix
Add 24ml UHP distilled water and leave for 2hr at room temperature
Add 48ml 0.2M Tris-HCl buffer, pH8.5
Final volume will be 200 mls
Stir with a clean stir bar on a hot plate on warm – heat gently to 50ºC stirring continually – at least 4-5 hours until the majority of the Mowiol powder goes into solution
Aliquot into 50 ml centrifuge tubes, weigh and balance
Centrifuge at 5000g for 15 minutes, there will be a pellet at the base, carefully remove supernatant
Aliquot into 15 ml conical tubes – add only 10 mls for expansion
Store at -20ºC for 12 months – store at room temperature no more than one month
This method is based, with permission, on an original protocol available here.