[3dem] plunge with vitrobot

[mf2944 at columbia.edu]

Dear Peter,

could you give a clarification on your suggestion?
Do you split the volume you normally add (eg 4 ul) onto both sides (eg  
2 ul left and 2 ul right), or do you apply the volume you normally use  
on both side (eg 4 ul left and 4 ul right)?

Cheers
Marcus



Message: 1
Date: Mon, 26 May 2014 22:08:00 +0000
From: Peter Shen <peteratbyu at hotmail.com>
To: Xiaofeng Fu <xiaofeng at brandeis.edu>, "3dem at ncmir.ucsd.edu"
         <3dem at ncmir.ucsd.edu>
Subject: Re: [3dem] plunge with vitrobot
Message-ID: <BAY168-W953631CCB071D9EB91C1E1C23B0 at phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"

Hi Xiaofeng,
I am familiar with this type of ice. It occurs when the droplet is  
poorly adsorbed across the grid surface. As Bob mentioned, depending  
on your particle, this type of suspension can be a great thing. I have  
found that applying the sample to both sides of the grid helps make  
this type of ice more uniform throughout the surface.
My two cents,Peter

Date: Sun, 25 May 2014 17:29:13 -0400
From: xiaofeng at brandeis.edu
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] plunge with vitrobot


Dear all,

Recently we had plunge problem by using vitrobot. Two images are  
attached: there is always a big droplet in the middle of square, and  
the ice is very thick at its neighboring area. We varied the GD from  
10s to 40s at 24mA, but the grids look similar. The buffer doesn't  
contain any salt or sugar.


Look forward to hearing any suggestions or experiences.

Thanks,
Xiaofeng


-- 
Marcus Fislage, PhD

Howard Hughes Medical Institute (HHMI)
Columbia University
Department of Biochemistry and Biophysics
Lab of Joachim Frank
New York, NY

Phone: 212.305.9532
Fax: 212.305.9500