Pelling:Protocols/Paraformaldehyde Buffer

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Fixative Buffer (2% Sucrose, 3.5% Paraformaldehyde - PFA)

  1. Place a 250mL Duran bottle in the fridge (or on ice) to keep it cold.
  2. In a 500mL beaker weigh out and add 2% Sucrose (by weight, 4 g), 3.5% PAF (by weight, 7 g).
  3. Add 200mL PBS and a magnetic stir bar.
  4. 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.
  5. The solution should turn clear after ~ 1 hour.
  6. 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.
  7. Label and store in fridge.

*** CAUTION: Do not inhale, work under fume hood ***