Creating Gelvatol Solution
Daniela Boassa, Mark Ellisman
Abstract
A gel-like mounting media (with an anti-fade ingredient) for fluorescence slides.
Before start
Add a small drop of Gelvatol to specimen.* Carefully place coverslip on the drop(s) avoiding bubbles.
- The mounting media will set overnight at 4 degrees or at room temperature 3-6 hours.
Steps
Add 2.4 g of Polyvinyl Alcohol (Mol. Wt. 30,000-70,000) to 6ml of glycerol.
Stir well to mix.
Add 6ml of double distilled water and leave for at least 2 hours at room temperature (rotate overnight)
Add 12ml of 0.2M Tris (pH 8.5).
2.4 ml Tris 1M
9.6 ml H2O
Heat to 50° C for 10 minutes with occasional mixing.
After polyvinyl alcohol is dissolved, clarify by centrifugation (5000g = ~6800 rpm) for 15 minutes at 20° C.
Use the large tabletop 5810R Eppendorf centrifuge.
Collect supernatant liquid and measure amount collected.
Add DABCO to 2.5% as antifade medium:
2.5g DABCO / 100ml of liquid
pH to 8.6 with pH paper (the paper will disintegrate, so pipette onto paper rather than putting paper into container).
Aliquot in 0.5ml microcentrifuge tubes and store at -20° C.
Stocks of Gelvatol are supposed to be stable at room temperature for several weeks after thawing.
