[3dem] blotting quantifoil from copper side

[Mohamed Chami]

Dear Daniel,
We had also this problem with Quantifoil grids because the paper filter doesn't have contact with the sample when you blot from the back side. The carbon film of Quantifoil's grids is thicker than the carbon film of Tedpella's grids. 2 tricks you can try:
- Glow discharges the both sides of the grids
or
- Put the sample (3 or 4 ul) of the sample on the carbon side and put also a little droplet of sample on the other side (blotting side) just by touching the middle of the grid with the tip of the pipette in order to make a bridge between the sample and the blotting side.
Hope it will help
Best
Mohamed

Mohamed Chami, PhD
Bio-EM lab
Center for Cellular Imaging and NanoAnalytics (C-CINA)
Biozentrum, University of Basel
Mattenstrasse 26, 
CH-4058 Basel
Switzerland
Tel.: +41-61-387 32 24

Fax: +41-61-387 39 86
E-mail: mohamed.chami at unibas.ch<mailto:mohamed.chami at unibas.ch>
http://www.c-cina.unibas.ch


________________________________
From: 3dem [3dem-bounces at ncmir.ucsd.edu] on behalf of Levy Daniel [Daniel.Levy at curie.fr]
Sent: Wednesday, January 20, 2016 4:29 PM
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] blotting quantifoil from copper side

Dear all
We have some difficulties to get homogenous ice when we blot our quantifoil grids with a Leica EM GP. We have low concentrated samples and we want to blot from the side opposite to the drop (the copper side). Our conditions worked well with holey formvar Ted Pella grids but not with quantifoil. We tried different grades of paper (1 or 4)  blotting conditions and glow discharged conditions. Any advice will be helpful.

Best
daniel

Daniel Lévy
UMR CNRS 168 Institut Curie
11 rue P.M.Curie
F-75231 Paris Cedex 05
France
Phone 33 1 56 24 67 82
Fax 33 1 40 51 06 36
http://umr168.curie.fr/en/Levy-group

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