[3dem] Problems blotting the liquid ethane
dokland at uab.edu
Fri Feb 22 10:34:31 PST 2008
There should be no need to remove the ethane after plunging. Although
it typically does solidify on the grid, it evaporates during the
transfer. I am curious where you got this procedure from -- at least
I have not heard of it before.
On Feb 22, 2008, at 11:50 AM, Eduardo Sanz-Garcia wrote:
> Hi all,
> I am having real problems to blot successfully the liquid ethane
> from my samples after freezing.
> I am using a vitrobot for freezing. This is very neat because makes
> everything automatic except the blotting of the liquid ethane.
> I know that I should see a drop of ethane in the filter paper, but
> this step has been very irregular for me (very few times I see
> filter paper wet).
> Do you have any suggestion for me?
> Does the speed of the process affect (remove the sample from the
> ethane, blot and put in the cryo box) the rate of success?
> Could be that the temperature around the filter paper is too low
> (nitrogen temperature) and that the ethane solidifies before
> touching the filter paper?
> For how long should I precool the filter paper?
> Thank you very much.
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