In this section:
- Bouin’s Fixative
- Dent's Fix (Methanol/DMSO)
- Paraformaldehyde(PFA), 20%
- PTw Containing 4% Paraformaldehyde
MEMFA (MOPS/EGTA/Magnesium Sulfate/Formaldehyde Buffer)
|1X||10X||for 1 L 10X|
|MOPS (pH 7.4)||0.1 M||1.0 M||209.27 g|
|EGTA||2 mM||20 mM||7.608 g|
|MgSO4||1 mM||10 mM||2.46 g|
- Prepare a 10 X solution without the formaldehyde for storage. Add fresh formaldehyde(1/10 volume of standard 30% stock) prior to use. The solution will turn yellow with age and when autoclaved.
For 50 ml: 35 ml Piric acid 12.5 ml Formaldehyde 2.5 ml glacial acetic acid Adjust volume, store at RT.
Dent's Fix (Methanol/DMSO)
- 4 volumes methanol
- 1 volume DMSO
Paraformaldehyde (PFA), 100mls of 4%
Measure 4g of Paraformaldehyde in the hood then add to 100 mls of pre-heated PBS.
Heat to around 62-64C.
Once dissolved aliquot 20 mls per tube and store at -20C.
PTw Containing 4% Paraformaldehyde
- 1 volume of 20% paraformaldehyde
- 4 volumes of PTw
Note: To made 20% paraformaldehyde in water, neutralize the cloudy solution with NaOH and then heat at 65 deg C, with shaking, until clear, Store at -20 dec C.
1X PBS(phosphate buffered saline)