recipe for 10% PFA: add 25g PFA to 250ml water . Heat at 50 degrees till dissolve.
diluting 10% PFA to 4% PFA: 10ml of 10% PFA + 2.5ml 10X PBS + 12.5ul of 20% triton + 12.375ml H2O.
sometimes i notice that my cells morphology look more rounded than normal. I was wondering if the fixation formula i use contains too much water such that water enters the cell (osmolarity issues), But i did dilute the 10% to 4% PFA using PBS. However, the 10% PFA is made entirely from water.
Could anyone look into this and suggest if i should increase/decrease the PBS concentration during dilution?
Swarnendra Singh
Hi Iwan,
-It must be direct 4% preparation.
-Freshly prepared for better results
-IHC grade chemical must be used
-Try not to heat, try to stir in warm PBS.
Best,
S
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