[3dem] Quantifoil grids
Mariena Silvestry Ramos
msilvestry at nysbc.org
Fri Dec 14 12:13:22 PST 2012
Dear Isabelle:
Greetings from NYC. I hope all is well. When I was in grad school we
used to order those grids. If I recall correctly, we used to do a wash
with acetone, a wash (or two with chloroform), a wash with ethyl acetate
and a wash with diethyl ether (though I'm not sure about that one). We'd
set the grids on top of a filter paper, in a glass petri dish, add a few
drops of each solvent, in that order, one after each round of the
previous solvent had dried. Used to take about a day or so. We also
baked them, which should've helped getting rid of any residual plastic.
But that plastic was indeed a pain to dissolve. Susan Saban at
Vanderbilt showed me that protocol, perhaps she can offer further insight.
Good luck!
Mariena
PS. Now we purchase C-coated grids directly from QF that come pre-coated
with carbon, no plastic. They work just as well and eliminated the
hassle of having to dissolve the plastic and waiting. Alternatively,
protochips has the C-flat grids that, from what I understand, may have a
lighter coat of carbon compared to QF, and also lack the plastic. Hope
this helps. Happy holidays
On 12/14/2012 2:55 PM, 3dem-request at ncmir.ucsd.edu wrote:
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> Today's Topics:
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> 1. Re: request (Steven Ludtke)
> 2. Quantifoil grids (Isabelle Rouiller)
> 3. Re: request (Steven Ludtke)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Fri, 14 Dec 2012 13:31:47 -0600
> From: Steven Ludtke<sludtke at bcm.edu>
> Subject: [3dem] Re: request
> To: "Heller, William T."<hellerwt at ornl.gov>
> Cc: "3dem at ncmir.ucsd.edu"<3dem at ncmir.ucsd.edu>
> Message-ID:<65683094-984C-478D-9C46-A9F3474C1EFE at bcm.edu>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi William. SAXS has been used with Cryo-EM quite a bit, though even that has fallen off in recent years as CryoEM resolutions have gotten down to 5 Å or so. SANS hasn't been used much as far as I can recall (or am aware). One of the benefits of SAXS with CryoEM is that hard X-rays and electrons scatter very similarly, making direct comparisons more straightforward. However, as you say, there may be potential applications where SANS could be quite useful in probing things we can't easily probe with electrons. I don't mind chatting on the phone about it, but I'm not sure how much I can offer personally. What I'd actually suggest is the 3dem mailing list, which reaches virtually the entire cryo-EM community : "3dem at ncmir.ucsd.edu"<3dem at ncmir.ucsd.edu>
>
> People make announcements about this sort of thing fairly frequently, so I don't think anyone is likely to resent your message. You will have to join the list (it's pretty low-bandwidth, usually a few messages a week), before you can send messages, as people may reply to you via the list, rather than directly. Just make sure to explain a little about SANS and its capabilities as part of the message. You can always drop membership after giving people a couple of weeks to respond...
>
> If you want to chat, as a practical matter, we may have to wait until the new year. I'm leaving for x-mas holidays early next week, and tied up in meetings/events almost continuously until then...
>
> cheers
> (and Merry X-mas)
>
> On Dec 14, 2012, at 12:09 PM, "Heller, William T."<hellerwt at ornl.gov> wrote:
>
>> Steve,
>> I hope that all is going well for you at BCM. Huey Huang visited ORNL recently and it certainly sounded like that was the case.
>>
>> The Biology and Soft Matter Division of the Neutron Sciences Directorate at ORNL is seeking to expand its portfolio of for neutron scattering in the biological sciences. To that end, we are contacting research groups that could potentially benefit from the facilities at ORNL. While I realize that there is unlikely to be strong interest from you specifically, I was hoping that you would have time for a chat regarding potential members of the virology and/or EM communities who might benefit from complementary data from neutron scattering experiments, such as SANS for improving models of RNA packing in the core of virions or from non-destructive in situ studies of virion structural changes. I thought that your position as co-director of the NCMI might have put you into contact with a fairly large community of researchers.
>>
>> Would you be willing to speak briefly, and would you have time next week before the holidays?
>>
>> Sincerely,
>> William
>>
>> William T. Heller, Ph.D.
>> Biology and Soft Matter Division
>> Neutron Sciences Directorate
>> EQ-SANS Lead Instrument Scientist
>> Oak Ridge National Laboratory
>> PO Box 2008; MS-6473
>> Oak Ridge, TN 37831
>>
>> phone 865-241-0093
>> fax 865-574-8363
>> email hellerwt at ornl.gov
>>
>> Adjunct Associate Professor
>> Department of Molecular& Structural Biology
>> North Carolina State University
>>
>>
> ----------------------------------------------------------------------------
> Steven Ludtke, Ph.D.
> Professor, Dept of Biochemistry and Mol. Biol. (www.bcm.edu/biochem)
> Co-Director National Center For Macromolecular Imaging (ncmi.bcm.edu)
> Co-Director CIBR Center (www.bcm.edu/research/cibr)
> Baylor College of Medicine
> sludtke at bcm.edu
>
>
>
>
>
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> Message: 2
> Date: Fri, 14 Dec 2012 19:38:41 +0000
> From: Isabelle Rouiller<isabelle.rouiller at mcgill.ca>
> Subject: [3dem] Quantifoil grids
> To: "3dem at ncmir.ucsd.edu"<3dem at ncmir.ucsd.edu>
> Message-ID:
> <0DD5A53003888247AF0EC8A711EE3602018CB97E at exmbx2010-9.campus.MCGILL.CA>
>
> Content-Type: text/plain; charset="us-ascii"
>
> Does anyone have a working protocol to remove the plastic from inside the holes of Quantifoil grids?
>
> Washing with chloroform does not work with the ones we presently have. We also tried flash freezing (it sometimes cracks the plastic, but not always). The distributor (SPI) has been unwilling to exchange them.
>
> Thank you,
>
> Isabelle
>
>
> Isabelle Rouiller, PhD
> Cell Biology Graduate Program Director
> Department of Anatomy and Cell Biology
> Room 115, Strathcona Anatomy and Dentistry Building
> 3640 University Street Montreal, Quebec H3A 2B2
> (514) 398-3244
>
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> Message: 3
> Date: Fri, 14 Dec 2012 13:55:32 -0600
> From: Steven Ludtke<sludtke at bcm.edu>
> Subject: [3dem] Re: request
> To: "3dem at ncmir.ucsd.edu"<3dem at ncmir.ucsd.edu>
> Message-ID:<5CB4A811-7792-4996-B704-8FF938AF1940 at bcm.edu>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi all. I will compound my mistake by sending another message apologizing for accidentally sending a private reply to the 3dem list. Regardless, William is looking for people who might be able to make use of SANS facilities, possibly for coordinated cryo-EM experiments. I'm sure he will forward another message himself with more details if he feels that didn't cover it :^)
>
> Apologies for filling your mailboxes. Happy Holidays
>
> ----------------------------------------------------------------------------
> Steven Ludtke, Ph.D.
> Professor, Dept of Biochemistry and Mol. Biol. (www.bcm.edu/biochem)
> Co-Director National Center For Macromolecular Imaging (ncmi.bcm.edu)
> Co-Director CIBR Center (www.bcm.edu/research/cibr)
> Baylor College of Medicine
> sludtke at bcm.edu
>
>
>
>
>
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> End of 3dem Digest, Vol 64, Issue 9
> ***********************************
--
Mariena Silvestry Ramos, PhD
Scientist - CEM Lab
New York Structural Biology Center
212-939-0660 ext. 9322
msilvestry at nysbc.org
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