A semi-permanent mounting medium for immunofluorescence microscopy

A semi-permanent mounting medium for immunofluorescence microscopy

Materials

• 6gm glycerol

• 2.4gm mowiol

• 6ml distilled water

• 12ml Tris buffer 0.2M at 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 53¡C until the mowiol dissolves, stirring occasionally. 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. Small amounts of DABCO or other fluorescence stabilizer (anti-fade agent) added to each aliquot immediately before use will reduce signal fading under UV light.