[3dem] Graphene oxide grids for negative stain

Fri May 27 13:23:33 PDT 2016

Dear Sanchaita,

we had really good success with the graphene oxide grids in cryo but only tried them once for negative stain. When we tried them for negative stain it was quite bad, with almost all grid squares broken. I am not sure if it was the graphene oxide or just bad luck. I would say in most cases carbon support is working fine for negative stain. 

Cheers,
Thomas

> On 27 May 2016, at 18:24, Ludovic Renault <llrrenault at GMAIL.COM> wrote:
> 
> Dear Sanchaita,
> 
> You might want to check a nice video made by Thomas Martin from the LMB in Sjors Scheres group.
> https://figshare.com/articles/Graphene_Oxide_Grid_Preparation/3178669 <https://figshare.com/articles/Graphene_Oxide_Grid_Preparation/3178669>
> 
> It's self-explanatory.
> 
> Cheers,
> Ludo
> 
> 
> On Fri, May 27, 2016 at 6:42 PM, Sanchaita Das <sanchaita06 at gmail.com <mailto:sanchaita06 at gmail.com>> wrote:
> Dear All,
> Has anyone tried Graphene oxide grids for negative staining of small proteins(<100KDa).
> If yes any tips and suggestions would be highly appreciated.
> Do you use normal Cu grids or holey carbon ones?
> Do you thin coat your grids with carbon or formvar before adding the Graphene?
> Regards,
> Sanchaita
> 
> On Thu, May 26, 2016 at 3:00 PM, <3dem-request at ncmir.ucsd.edu <mailto:3dem-request at ncmir.ucsd.edu>> wrote:
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> Today's Topics:
> 
>    1. Re: Old camera and scanner to give out (kunihiro uryu)
>    2. Research Scientist Position in the Simons Electron Microscopy
>       Center at the New York Structural Biology Center (William Rice)
> 
> 
> ----------------------------------------------------------------------
> 
> Message: 1
> Date: Thu, 26 May 2016 08:50:59 -0400
> From: kunihiro uryu <kuryu at mail.rockefeller.edu <mailto:kuryu at mail.rockefeller.edu>>
> To: Zongli Li <zongli.li at gmail.com <mailto:zongli.li at gmail.com>>
> Cc: 3DEM Mailing List <3dem at ncmir.ucsd.edu <mailto:3dem at ncmir.ucsd.edu>>
> Subject: Re: [3dem] Old camera and scanner to give out
> Message-ID:
>         <CAFK2HhbzsnV52OrN6wLK7Dv9YhDZKsptQvHz5OYUya+AHx1z1Q at mail.gmail.com <mailto:CAFK2HhbzsnV52OrN6wLK7Dv9YhDZKsptQvHz5OYUya%2BAHx1z1Q at mail.gmail.com>>
> Content-Type: text/plain; charset="utf-8"
> 
> Hi Zongli,
> 
> Thank you for letting me know. It was a great gift. I totally understand.
> 
> Regards,
> Hiro
> ------
> Kunihiro Uryu, Ph.D.,
> Research Assistant Professor
> Director of Electron Microscopy
> Resource Center (EMRC)
> RRB Rm120
> The Rockefeller University
> 1230 York Ave., Box 230
> New York, NY 10065
> 
> On Wed, May 25, 2016 at 7:28 PM, Zongli Li <zongli.li at gmail.com <mailto:zongli.li at gmail.com>> wrote:
> 
> > Hello everyone,
> >
> > Thanks a lot for your response. The two CCD cameras have been taken
> > already. But
> > two scanners are still available.
> >
> > Best regards,
> >
> > Zongli
> >
> > On Wed, May 25, 2016 at 5:08 PM, Zongli Li <zongli.li at gmail.com <mailto:zongli.li at gmail.com>> wrote:
> >
> >> Sorry, I made a mistake when sending out previous email regarding the CCD
> >> camera:
> >> Here is the correct message:
> >>
> >> 1. 1024x1024 Gatan MultiScan CCD, pixel size 24 um, model 794.
> >> 2. 2048x2048 Gatan MegaScan CCD, pixel size 30 um (4k chips). Model
> >> 794/20.
> >> 3. Imaging plate scanner (DITABIS Micron scanner, Germany).
> >> 4. SCAI scanner, used to digitize the negatives.
> >>
> >> Best regards,
> >>
> >> Zongli
> >>
> >> On Wed, May 25, 2016 at 4:09 PM, Zongli Li <zongli.li at gmail.com <mailto:zongli.li at gmail.com>> wrote:
> >>
> >>> Dear colleagues,
> >>>
> >>> We have following items we don't use anymore and would like to give them
> >>> out if someone can still use them:
> >>>
> >>> 1. 1024x1024 Gatan CCD camera, megascan, with 2k chips (pixel size 30 um)
> >>> 2. Imaging plate scanner (DITABIS Micron scanner, Germany).
> >>> 3. SCAI scanner, used to digitize the negatives.
> >>>
> >>> They are all in good working conditions.
> >>>
> >>> If interested, please contact me directly at zongli_li at hms.harvard.edu <mailto:zongli_li at hms.harvard.edu>.
> >>> You will be responsible for arranging the shipment.
> >>>
> >>> Best regards,
> >>>
> >>> Zongli
> >>>
> >>
> >>
> >
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> ------------------------------
> 
> Message: 2
> Date: Thu, 26 May 2016 11:28:51 -0400
> From: William Rice <rice at nysbc.org <mailto:rice at nysbc.org>>
> To: 3dem at ncmir.ucsd.edu <mailto:3dem at ncmir.ucsd.edu>, ccpem at jiscmail.ac.uk <mailto:ccpem at jiscmail.ac.uk>
> Subject: [3dem] Research Scientist Position in the Simons Electron
>         Microscopy Center at the New York Structural Biology Center
> Message-ID: <57471633.8060803 at nysbc.org <mailto:57471633.8060803 at nysbc.org>>
> Content-Type: text/plain; charset="utf-8"; Format="flowed"
> 
> Research Scientist Position in the Simons Electron Microscopy Center at
> the New York Structural Biology Center
> 
> The New York Structural Biology Center (NYSBC) seeks an experienced
> electron microscopist to join the staff of the Simons Electron
> Microscopy Center (http://semc.nysbc.org <http://semc.nysbc.org/>). The NYSBC is a shared center
> that supports state-of-the-art research in EM, NMR, and X-ray
> crystallography. The facilities include five transmission electron
> microscopes (including 2xTitan Krios) and a dual-beam scanning electron
> microscope (with a Quorum cryo stage), which support projects involving
> electron tomography, single particle analysis and electron
> crystallography of both stained and frozen-hydrated samples. Projects
> focus on 3D reconstruction of biological assemblies ranging from the
> atomic structure of membrane proteins, to the subunit organization in
> macromolecular complexes and the cellular anatomy of developing
> organisms, to 3D reconstructions of entire cells and their distributions
> within their native tissue using the dual-beam microscope.
> 
> To assist in these developments, NYSBC seeks an individual with
> postdoctoral experience in biological electron microscopy and image
> reconstruction. This individual will carry out experiments in support of
> collaborative projects with affiliated investigators and will also have
> opportunities to pursue independent research projects. The individual
> should be capable of multitasking, should enjoy working with other
> people, should have a good working knowledge of electron microscopes and
> a strong research background. Good communication skills are essential.
> 
> To apply, please send a c.v. and the names of three references to
> emg at nysbc.org <mailto:emg at nysbc.org> <mailto:emg at nysbc.org <mailto:emg at nysbc.org>>.
> 
> --
> William Rice, Ph.D.
> Senior Scientist
> New York Structural Biology Center
> 89 Convent Ave.
> New York, NY 10027
> (212) 939-0660 x344 <tel:%28212%29%20939-0660%20x344>
> 
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> End of 3dem Digest, Vol 105, Issue 23
> *************************************
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> 
> -- 
> "For one human being to love another! That is perhaps the most difficult of all tasks...The work for which all other work is but preparation"
> Rainer Maria Rilke
> 
> 
> Sanchaita Das
> 
> 
> 
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