[3dem] Thinner ice after several microscope uses
Eduardo Sanz Garcia
esanzgar at gmail.com
Mon Nov 24 18:29:49 PST 2008
I was analyzing the success rates of my cryoEM sessions and I noticed a
trend: When I am the first microscope user of the day I always have
thick ice and the chances to obtain better ice improve after several
consecutive trials.
First user of the microscope (No previous users before me)
Number of days with good ice
Number of days with thick ice
First trial
*0*
*11*
Second trial
5
3
Third
1
1
It could be either:
* I don't pre-cool well the cryoholder and the first use is not
totally ready (I cool off the holder inside the microscope, and it
is always at -180 C)
* Microscope related issues (the vacuum is better after sometime,
*we start a cryocycle every night*).
I believe that it is a microscope related issue because, when I used the
microscope after another user I get better chances to obtain thin ice:
*After* another microscope user
Number of days with good ice
Number of days with thick ice
First trial
7
3
Have anyone noticed a similar trend?
Additionally, I would like to ask you how consistent your sample
preparation are.
I always prepare the samples in the same way, and I get very
inconsistent results, sometimes thick ice, sometimes thin ice,
If you prepare several sample under the same conditions (vitrobot,
handling, etc...) , do you obtain
* 100% consistent results
* 80-90% consistent results
* less than 50% consistent results?
Thank you very much.
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