Does anyone know that how long can fixed mouse brain slices be stored in 4C in PBS? Or can these samples still be used for immunostainning? We perfused the mouse with PBS and 4%PFA, post-fixed in 4%PFA overnight, then frozen cut the slices into 40um. the longest one has been put there for around 1 YEAR.
It should depend on what you are looking for.
If you find no fungal/bacterial growth and no visible degradation, then you can perform your immunostaining where appropriate negative and positive controls are included. You should find your answer there.
Hi Yubo,
Storing the tissue in PBS for a long time can increase the chance of bacterial growth in your buffer. Bacteria produce proteases that can break down the proteins in your tissues. Also, during that long storage, the water can evaporate, and the salt concentrations increased. The best way to store them is: after fixation switch to 50% and then 70% ethanol (at 4C). You can store the tissue in ethanol for a few months. However, long-term storage in ethanol can also be problematic, as, by the time, the cells lose more water and become more hydrophobic. This makes the process of antigen retrieval more difficult.
Best wishes and good luck,
Reza
If you want to store for long term for immunohistochemistry, better not to store in PBS as the chances of bacterial growth are very high. I recommend to store them in 70% ethanol for long term but keep on changing the solution after each month or if you see it turbid.
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