Fixatives

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In this section:

MEMFA
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
Formaldehyde 3.7%
  • 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.


Bouin’s Fixative

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), 20%

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.


PTw

1X PBS(phosphate buffered saline)

0.1% Tween-20