Hi all,
I wanted to know if snap freezing the live tissue is helpful when one is aiming for paraffin embedding and sectioning (at room temp) later. Will the tissue gets damaged when it thaws?
My reason to go for the above approach is that putting the tissue directly in fixative which is generally toxic might alter the cell metabolism and lead to cytotoxic death.
I know the method depends on what you want to look at. It will be great if someone can suggest for various scenarios like structural/architectural studies, immuno staining etc
Paraffin embedding will require fixing your tissues even after snap freezing to prevent them from decaying. Therefore, you won't really gain much (if anything) from snap freezing the tissue rather than immersing it immediately in fixative.
If you are concerned about changes in cellular metabolism during the fixation process, you can quickly slice your tissue into thinner sections before fixing.
For analysis of tissue architecture and immunostaining, formaldehyde fixed paraffin embedded tissue (FFPE) is the gold standard for most tissue types.
Hi, if your aim is to perform paraffin embedding, just place your tissue into 10% buffered formalin and this will preserve/fix you tissue in preparation for processing at a latter stage. Snap freezing can cause cellular damage and degrade the benefits of paraffin based Microscopy.
The best way to obtain good histological sections is to fix the fragments in formalin immediately after sampling. In your case freezing will be more useful for the study of enzymatic activity, but after thawing you can obtain good paraffin samples if you work fast but the frozen water can also cause cellular damage!
Thanks all for your responses. I work with Drosophila by the way. not larger tissues like that of mice or human.
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