I would like to know if anyone knows what happens if the timing of fixation with p-formaldehyde exceed 48 h and if is reversible the excessive hardening which is obtained mainly at the level of the wall of gram-negative.
Thank you in advance!
Gudrun Lang
For FFPE tissue we do a permeabilization step before digestion. If you can immobilizise your cells on an adhesive slide, you can try this.The sections are immersed in hot citrate-buffer pH 6 for 60 min at 85°C. Maybe cells don't need as long as this.
Permeabilization and digestion have a relation. the more of the first, the less of the second.
This hot-buffer step is like the antigen-retrieval in IHC.
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