Preparation and use of the O.T.E.
The preparation of the OTE solution is similar to the infusion of tea water 70°C or Buffer 0,1M.(*)
– Using a heating magnetic stirrer set at 70°C, dissolve your OTE powder at a concentration of 0.2% (because saturation is reached at 0.3%)
– After complete dissolution (15 to 30 minutes with stirring at 70 ° C),
- Filter your OTE extract solution at 0.45µ.
- Store in a small Eppendorf tube filled to the maximum to reduce air containing oxygen or in an AirLess bottle.
- Store in the darkness at 4 °C (when handling always let your solutions return to RT)
- Contrast your grids with OTE for 15 to 25 minutes, darkness, RT
- Drain simply not rinse
- Post-stain 1 to 2 minutes with lead citrate according to Reynolds (Ref # 11300 from Delta Microscopies)
- Drain, short rinse, dry and ready to watch.
*note: the solution can be kept for no more than a week, but more time in a cacodylate buffer solution, AirLess bottle (no bacterial growth).