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How to Make 4% Paraformaldehyde (4%PF)

Make 4% PF 1000ml:

1. Fill a 2000ml Beaker with about 400ml H₂O, put a thermometer into the beaker. Heat beaker at hot stir plate to reach temperature at 62-64°C.
2. 980ml RNAse free PBS in 1000ml bottle. Put a stir bar into PBS bottle. Add 100ul 10N NaOH (7 drops 10N NaOH) and 40g paraformaldehyde into PBS.
3. Put bottle into beaker and stir the paraformaldehyde.
4. Watch the temperature while stiring the paraformaldehyde. When the temperature reach to 62-64°C, the fixative will be clear gradually.
5. Remove fixative from hot plate, cool down to room temperature.
6. Filte fixative.
   The pH of this fixative should be around 7.3; check pH is not necessary.
7. Aliquot fixative and store at -20 °C.

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