[Histonet] Cutting fresh-frozen brains from 1 week old rat pups

[Adam Galle]

Hi all,
Currently I am working with brains from 7 day old rat pups, that undergo an 
hypoxic-ischemic injury (Levine/Vannucci technique). These brains are 
unfixed, frozen in isopentane and cut at 20um on a croystat. These brains 
are not cutting very well compared to an adult brain (potenially due to 
unfinished myelination?), they are very 'crumbly' for want of a better term 
and always have cracks or generally poor preservation of morphology. I have 
tried all the standard tricks of different temperatures, section thickness 
and knife angle to no avail. I am going to perfuse fix my next cohort of 
animals with PFA and then a 30% sucrose step to see if that helps, but I was 
hoping that someone out there would have some tips on cutting these immature 
brains.

Thanks,
Adam.


Adam Galle,

Neuropharmacology and Brain Injury Lab
Department of Pharmacology
School of Medical Sciences
UNSW Sydney