I have been preparing 10% EDTA solution for decalcification of rat knee joints that I want to use for IHC, and have been using it for 2.5 weeks. However, even though the pH may only change by 0.2 units, I am wondering if it would be better to complete the process using 10% EDTA in PBS to provide a buffer. Would this affect the cell morphology of the tissue if I switch from water to PBS?
PBS has the osmotic pressure of normal cells. If your tissue was in H2O this means at the beginning the buffer will be hypertonic, but in my experience it has no effect on whole mount embryos unlike hypotonic buffer which can lead to rips in the epithelium due to osmotic pressure. But if you want to be save you could always do an extra step with half volume old buffer, half new.
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