[3dem] bursting cells grown on grids

[Julika Radecke]

Hi Michael,

have you tried using blotforce 0 or higher for blotting cells open with a vitrobot? Or blotting with a different type of blotting paper, or repetitive blotting?
I have always used the Vitrobot for freezing primary neurons, at blotforce -10 they stay intact. My impression is though that cells that were about to die will burst at -10, so I am thinking at 0 or higher blotforces they might burst.

Best,
Julika
________________________________
From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Michael Elbaum <michael.elbaum at weizmann.ac.il>
Sent: 21 October 2024 23:14:18
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] bursting cells grown on grids

Hi all,     the Vitrobot used to have a reputation for bursting cells grown directly on grids. I didn't find any proper reference, though I'm aware of a "fix" to replace one of the filter papers by a teflon disk. In a current project we actually want to burst the cells open to leave organelles and cytoplasm exposed. Standard unroofing with a stream of buffer will wash it away. I wonder if anyone can share experience, even if frustrating, on the burst directly between two filter papers.
thanks,
Michael




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