[3dem] getting protein samples into holes in quantifoil

[Natesh Ramanathan]

Dear Sheemei,

These are due to the characteristics of the protein to stick to carbon.
We faced similar problem and one trick to overcome this problem is
use a thin layer are carbon on the holes of holey grid.  Another advantage
is you can reduce the concentration of sample required to a large
extent; you can try almost 1/10th that you use in holey grids.
 Use of plasma treated graphene  is another method that you can try -  see
recent Nat. Methods paper by Lori and group "Controlling protein adsorption
on graphene for cryo-EM using low-energy hydrogen plasmas. Russo CJ,
Passmore LA.  Nat Methods. 2014 Jun;11(6):649-52.

Hope that helps,
Warm Regards,
Natesh

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"Live Simply and do Serious Things .. "
- Dorothy Mary Crowfoot Hodgkin OM, FRS

"In Science truth always wins"
- Max Ferdinand Perutz OM FRS
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Dr. Ramanathan Natesh
Ramalingaswami Fellow-DBT, Assistant Professor
Indian Institute of Science Education and Research Thiruvananthapuram
(IISER-TVM),
1st floor, Dept. of Computer Science & Engg. Building,
CET Campus, Engineering College P.O.,
Trivandrum,  695016, Kerala, India

natesh at iisertvm.ac.in
http://www.researcherid.com/rid/C-4488-2008
http://iisertvm.ac.in/~natesh
Ph. 0091- 471-2599403
Fax.0091-471-2597427

On 31 October 2014 11:28, Sheemei Lok <sheemei.lok at duke-nus.edu.sg> wrote:

>  Dear all,
> Our protein sample tend to stick to the carbon film instead of the hole in
> quantifoil. Are there any tricks that we can do to get them into the holes?
>
>  Regards,
>  Shee-Mei Lok
>
>
>
>
>
>
>
>
>
> *Associate Professor Program in Emerging Infectious Diseases, Duke-NUS
> Graduate Medical School, KTP Building Level 9 Room 19 8 college Road,
> Singapore 169857 Ph: 65-65165840 Fax:65-62212529
> email: sheemei.lok at duke-nus.edu.sg*
>
>
>
>
>
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