[3dem] specimen preparation

[Kimberley Cheng]

Dear all,

I have difficulties preparing cryoEM specimens of my samples, and I would appreciate any help I can get. Both samples contain lipid nanoparticles, and I use a Vitrobot to prepare the frozen-hydrated specimens. The problems are the following

Sample 1: Majority of the grid has no ice at all. If I am lucky, a small region of the grid (1/4 of a grid square) is covered with ice that is very thick. What can be the problem? Any advice on how to improve the procedure?

Sample 2: Ice thickness and particle distribution is good. The concentration is, however, very low. Any suggestions on how to increase the on-grid particle concentration? I have already tried preparing the sample on continuous carbon instead of perforated carbon film.

I am very interested in hearing your opinions in this.

Thanks in advance,
Kimberley Cheng



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