Fixative Buffer (2% Sucrose, 3.5% Paraformaldehyde - PFA)
- Place a 250mL Duran bottle in the fridge (or on ice) to keep it cold.
- In a 500mL beaker weigh out and add 2% Sucrose (by weight, 4 g), 3.5% PAF (by weight, 7 g).
- Add 200mL PBS and a magnetic stir bar.
- Heat solution to ~65°C on a magnetic stirring hot plate in a fume hood, do not heat over 70°C or PAF will degrade. Cover beaker with tin foil and insert a thermometer through top to keep track of the temperature.
- The solution should turn clear after ~ 1 hour.
- After solution has turned clear, remove the magnetic stir bar and pour the hot liquid directly into the cold bottle in the fume hood and seal the bottle.
- Label and store in fridge.
*** CAUTION: Do not inhale, work under fume hood ***