SV: [3dem] Sucrose

Kimberley Cheng Kimberley.Cheng at ki.se
Fri May 27 07:58:35 PDT 2011


Thank you all for your feedback! And also for the reference. Your suggestions are gratefully accepted. Some of my samples contain up to 9% (w/v) sucrose, so hopefully using buffer exchange or washing with sucrose-free buffer will do the trick.

Thanks again!

All the best,
Kimberley

________________________________
Från: 3dem-bounces at ncmir.ucsd.edu [3dem-bounces at ncmir.ucsd.edu] för Sacha De Carlo [sachadecarlo at yahoo.com]
Skickat: den 27 maj 2011 11:25
Till: Kimberley Cheng; 3dem at ncmir.ucsd.edu
Ämne: Re: [3dem] Sucrose

Hi Kimberly,

I would avoid having more than 5% (w/v). At 15% you have almost no contrast anymore, that of course depends on the sample you are studying.

I would try to do a buffer exchange with trehalose. Of course, my humble opinion is heavily biased, but documented (De Carlo et al., 1999)

REF: De Carlo, Adrian, Kälin, Mayer and Dubochet (1999), Unexpected property of trehalose as observed by cryo-electron microscopy. Journal of Microscopy, 196: 40–45. doi: 10.1046/j.1365-2818.1999.00591.x

Best of luck,
Sacha


=============================
Dr. Sacha De Carlo
Sr. Res. Scientist
FEI Company
________________________________
From: Kimberley Cheng <Kimberley.Cheng at ki.se>
To: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
Sent: Friday, May 27, 2011 10:24 AM
Subject: [3dem] Sucrose

Dear all,

We are trying to analyze some samples that contain sucrose. Of course the best would be to not have any sucrose at all, but in these cases we have no choice since it's necessary for stability. My question is, does anybody know the acceptable level of sucrose in vitrified specimens? Any literature reference would be great.

All the best,
Kimberley


Dr. Kimberley Cheng
Karolinska Institutet, Department of Biosciences and Nutrition and
The Royal Institute of Technology, School of Technology and Health

Phone: +46-(0)8-608 92 17
Fax: +46-(0)8-608 92 90
E-mail: Kimberley.Cheng at .ki.se<http://ki.se>



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