Mowiol (Aqueous mounting media)

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

Method

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.

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