A water-based semi-permanent mounting medium for immunofluorescence microscopy

 

Materials

• 6g glycerol
• 2.4g mowiol 4-88
• 6 ml distilled water
• 12 ml 0.2M Tris buffer·Cl pH 8.5

Method

Weigh out the glycerol in a 50ml plastic tube and add the mowiol. Stir thoroughly. Add water and leave for 2hr at room temperature. Add the Tris buffer and incubate at approximately 50°C until the mowiol dissolves, stirring occasionally (this might take several hours). Make sure all mowiol has dissolved, other wise the refractive index (RI) of the medium will be lower. Clarify by centrifugation at 4000-5000 RPM for 20 minutes and aliquot into small tubes. Stored frozen it should last for up to12 months. The solution is stable at room temperature for 1 month. 2.5% of DABCO or other fluorescence stabilizer (anti-fade agent) added to each aliquot immediately before use will reduce photobleaching.

We strongly recommend to measure the actual RI of a test sample of the set medium and use this value for any further planning of your experiments or the data analysis.