[3dem] Unrecognised CryoEM contaminant 2
rkhayat at ccny.cuny.edu
rkhayat at ccny.cuny.edu
Fri Mar 28 10:00:29 PDT 2014
The hobgolins are the same size as the virus particles!
Perhaps treating the samples with DNase/RNase and seeing if
they disappear will test the possibility that they are DNA/RNA
and not hobgoblins...
Reza Khayat, PhD
Assistant Professor
The City College of New York
Department of Chemistry, MR-1135
160 Convent Avenue
New York, NY 10031
Tel. (212) 650-6070
---- Original message ----
>Date: Fri, 28 Mar 2014 09:46:59 -0700
>From: 3dem-bounces at ncmir.ucsd.edu (on behalf of Gabriel
Lander <glander at gmail.com>)
>Subject: Re: [3dem] Unrecognised CryoEM contaminant 2
>To: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
>
> It would help if you posted some images of the same
> sample in negative stain. Is there a possibility
> that this is condensed RNA? It would be almost
> invisible in negative stain, but show up very nicely
> in cryo.
> Otherwise I'm inclined to say that these are
> microscopic hobgoblins that have infested your
> plunging device.
> On Mar 28, 2014, at 9:42 AM, "Xinghong Dai"
> <bestdz at gmail.com> wrote:
>
> Hi Daniel,
> To me, it looks like phantom caused by too thin
> ice. Especially black patches around the hole in
> up-left panel, they look like almost-dry areas.
> Try to increase the humidity, and wait shorter
> time before, and after, you blot. Also try shorter
> blot time.
> Good luck!
>
> Best regards,
>
>
> Xinghong Dai
>
> *********: 3dem-bounces at ncmir.ucsd.edu [mailto:3dem-
bounces at ncmir.ucsd.edu] ****** Daniel
> Luque
> ************: Friday, March 28, 2014 5:52 AM
> *********: 3dem at ncmir.ucsd.edu
> ******: [3dem] Unrecognised CryoEM contaminant 2
>
> Hello
>
> Before a couple of days waiting for the list
> moderator approval to attach the images in the
> list , I do not have received any response (either
> positive or negative) and I do not have find a
> mail/way for contact. Thus, I have made the images
> available at our server at:
>
> http://halley.cnb.csic.es/~virus/3DEM/Example_images.pdf
>
> Bellow is text of the original question:
>
> We are doing CryoEM of viral particles. When we
> check the sample by negative staining, a field of
> intact homogeneous particles without significant
> contaminants is observed. However, when we
> visualize the same sample by CryoEM, additionally
> to a lower-than expected number of particles, we
> observe an unexpected contaminant mostly located
> in the holes (see attached document with some
> example images). Does anybody see this class of
> contaminants? Any idea about its origin?
>
> Thanks in advance!
>
>
> Daniel Luque
>
>
>
>
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