[3dem] tomography of plastic sections at N2 temperature

[Schwartz, Cindi (NIH/NIAID) [E]]

Hi Daniel,

>From experience garnered in the Boulder lab, this can work. The biggest problem you face is that the section still shrinks, just at a much slower pace. So you have to make sure you 'cook' the sample long enough so that the shrinkage does not occur in the first part of your tilt-series. You probably have to figure this out empirically.

In Boulder, we had 2 Tecnai machines and we did cryo and plastic interchangeably (sometimes even on the same day) with no visible effects on the scope or the samples. We always looked at the plastic sections at room temperature. However, now that I am at RML and we have a Krios, we keep plastic sections to our room temp scope and cryo to the Krios.

I have used a plastic section in the Krios to complete SerialEM calibrations. For that, I loaded the grid warm into the autoloader rings, loaded the cassette into the Krios warm (with warm autoloader) and then cooled everything down. I would worry that the flimsy carbon/formvar (especially across a slot grid) would break. However, someone might have better experience than me on that front.

Good luck!
Cindi



From: Levy Daniel [mailto:Daniel.Levy at curie.fr]
Sent: Wednesday, August 30, 2017 5:03 AM
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] tomography of plastic sections at N2 temperature

Dear All,
we will be interested by your advice, experience, opinion on the possibility to perform tomography on plastic sections (resin embedded, tokayasu) cooled at N2 temperature, e.g. within an cooled autoloader: -time for temperature stabilization, possible shrinkage of sections, contamination of the sample .. or if plastic section should be definitively imaged at RT ?
best
daniel
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