[3dem] 3dem Digest, Vol 95, Issue 16

[Nijpels Frank (ID)]

Hi Li,

What kind of plunger do you use?

regards,

Frank


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Van: 3dem [3dem-bounces at ncmir.ucsd.edu] namens 3dem-request at ncmir.ucsd.edu [3dem-request at ncmir.ucsd.edu]
Verzonden: donderdag 23 juli 2015 21:47
Aan: 3dem at ncmir.ucsd.edu
Onderwerp: 3dem Digest, Vol 95, Issue 16

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Today's Topics:

   1. vitrified ice but not continuously, How to make it better?
      (Li, Kunpeng)


----------------------------------------------------------------------

Message: 1
Date: Thu, 23 Jul 2015 10:27:09 +0000
From: "Li, Kunpeng" <li2182 at purdue.edu>
To: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
Subject: [3dem] vitrified ice but not continuously, How to make it
        better?
Message-ID:
        <1160C903B1262A488C0DAECFE601E1AC1C11E1 at WPVEXCMBX03.purdue.lcl>
Content-Type: text/plain; charset="us-ascii"


Dear All,

        Recently I want to optimize the freeze conditions of a protein complex, But No matter how I can change the conditions, I cannot get the thin and continuous vitrified ice. the mass of the protein complex is almost 900 kDa. and the carbon grid is Quantifoil 1/1, glow discharge time is 1.5 minutes, the humidity in the plunger chamber is more than 80%RH,  and the ice thickness I estimated is near to 60 nm, I post the images from search mode to photo mode under CM200 with 1k*1K CCD.  I tried a low salt buffer and longer blot time at this check round. The accurate number post on the attached images.
         Do you confused with these similar problems once before?  Could you give me some clues to improve it?

          Thanks for any help.



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