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

[Merced Leiker]

Definitely do at least that one sucrose cryopreservation step you 
mentioned.  Even step up to it in 10% and 20% sucrose.


--On Wednesday, March 18, 2009 4:04 PM +1100 Adam Galle 
<adam.galle <@t> student.unsw.edu.au> wrote:

> 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
>
>
>
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Merced M Leiker
Research Technician II
354 Biomedical Research Building
School of Medicine and Biomedical Sciences
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214
Ph: (716) 829-6033
Fx: (716) 829-2725

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