I am performing a set of intracerebral injections and I would like to perform an assay to determine neuronal damage around the injection site. Following the injections, mice are perfused and fixed in PFA 4% and brains are sectioned in a freezing microtome (50 um).
I am concerned that the injected solution or the large volume (1 ul) might produce damage leading to, for example, apoptosis, necrosis or even milder cellular damage.
Hi,
Have a look at this article:
Article Osthole confers neuroprotection against cortical stab wound ...
:)
You could just perform a TUNEL IHC. Roche has a great kit for this.
I suggest that, whichever the method you use to target defined mechanisms of cell death (such as TUNEL, directed at DNA breaks), you examine the tissue around the lesion in sections stained with good old conventional methods. It may sound outdated to stain sections with a hematoxylin/eosin protocol, but it will give you important information, often of more value than limited scope methods.
Dear Valeria, Renee and Rafael.
Thanks a lot for your answers! It has been very helpful!
Cheers
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