The younger the animal is, the more the brain will "bend" as you
approach it with the blade. Also, the bigger the brain block (taller
in jonesmat's words), the more it will bend.
What kinda of vibratome are you using? You want your vertical
displacement to be as large as possible (the is changeable on the
VT1000). Perhaps you need to advance the blade more slowly? Or
increase the vibration speed. Don't be afraid to change the settings
from what the "manufacturer recommends".
The cutting chamber is at 4 degrees or colder right? The warmer the
buffer, the more wiggly the brain gets.
Preperfusion wont help you with your problem. All perfusion does, in
my experience, is to improve the optics of the slice, i.e. you can see
I would avoid mounting the brain in agar if at all possible. I only
use that for things like brain stem slices, or retinal slices.
On Mar 4, 5:09 pm, Dhruba Pathak <pathakdhr... from gmail.com> wrote:
> Dear all,
> It would be nice if some of the colleagues has performed bolus drop
> method of perfusion and could share their experience. I would like to
> try this method of perfusion protocol. But, I have not find the
> details yet. Would anyone like to suggest where can I find the
> For example, what should be the size of drop of test substance? In
> which angle the test substance should be drop? And, whether it is
> necessary to drop directly above the cell? What the volume of
> physiological saline should be present in experimental chamber while
> dropping substance?
>> Thank you, and hope to get some idea.
> Dhruba Pathak
> PhD student,
> Institute of Pathological Physiology
> Universtiy of Belgrade
> Belgrade 11000