<div dir="auto">Hi Cindy, <div dir="auto">Try to dissolve plastic with THF (Tetrahydrofuran). The only important tip is to move fast because it evaporates very quickly</div><div dir="auto">Good look</div><div dir="auto">Pedro</div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">El lun, jul 1, 2019 11:37, <<a href="mailto:3dem-request@ncmir.ucsd.edu">3dem-request@ncmir.ucsd.edu</a>> escribió:<br></div><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex">Send 3dem mailing list submissions to<br>
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Today's Topics:<br>
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1. Re: Help! Plastic films on my holey grids are KILLING ME<br>
(Lars-Anders Carlson)<br>
2. Re: Help! Plastic films on my holey grids are KILLING ME<br>
(Mariena Silvestry Ramos)<br>
<br>
<br>
----------------------------------------------------------------------<br>
<br>
Message: 1<br>
Date: Mon, 1 Jul 2019 19:23:10 +0200<br>
From: Lars-Anders Carlson <<a href="mailto:lars-anders.carlson@umu.se" target="_blank" rel="noreferrer">lars-anders.carlson@umu.se</a>><br>
To: <<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem@ncmir.ucsd.edu</a>><br>
Subject: Re: [3dem] Help! Plastic films on my holey grids are KILLING<br>
ME<br>
Message-ID: <<a href="mailto:7aedd66e-c28f-40a6-769c-d8db322c0ab4@umu.se" target="_blank" rel="noreferrer">7aedd66e-c28f-40a6-769c-d8db322c0ab4@umu.se</a>><br>
Content-Type: text/plain; charset="windows-1252"; format=flowed<br>
<br>
Hi Cindi,<br>
Can you remove it by just plasma cleaning longer? That could be the <br>
reason why some of us typically don't see this film in the first place <br>
(another reason could of course be that we don't look as closely as you).<br>
<br>
QF grids used to have plastic deposits around the edge of the holes, <br>
which is where they would start bubbling at higher exposures (80-100 <br>
e/?2), but I haven't seen much of this lately.<br>
<br>
I hope you find a solution since you seem really frustrated about it.<br>
<br>
Cheers,<br>
Lars<br>
<br>
On 2019-07-01 18:38, Schwartz, Cindi (NIH/NIAID) [E] wrote:<br>
> Hi all,<br>
> <br>
> We?ve been suffering with trying to get the plastic films off our <br>
> Quantifoil grids with very inconsistent results. We?ve also had film <br>
> issues on the c-flats, but my understanding is that the Quantifoils use <br>
> a plastic background and the c-flats use a carbon background. Please <br>
> correct me if I?m wrong on that one. We use both copper and gold grids <br>
> with the carbon substrate of various different patterns from R1/4 to <br>
> Multi-A with grids that we purchased 2 years ago to ones we?ve received <br>
> last month.<br>
> <br>
> The list of solvents we?ve tried that should dissolve plastic films:<br>
> <br>
> Ethyl acetate<br>
> <br>
> Chloroform<br>
> <br>
> Ethylene dichloride<br>
> <br>
> Trichlorobenzene<br>
> <br>
> The list of solvents to clean them up a bit:<br>
> <br>
> Acetone<br>
> <br>
> Ethanol<br>
> <br>
> 5% acetic acid<br>
> <br>
> We?ve tried sonication (a disaster)<br>
> <br>
> Submerging in drops<br>
> <br>
> Submerging in vast quantities of liquid<br>
> <br>
> With/without little stir bars while the grids are on a metal platform in <br>
> the solvent<br>
> <br>
> Time ranges from 15 min to overnight<br>
> <br>
> With/without a light glow discharge before cleaning<br>
> <br>
> We?ve put 10 nm colloidal gold fiducials on both sides of the grids <br>
> (like you?d use for plastic section tomography?where they stick to <br>
> Formvar just fine) after cleaning with various permutations from the <br>
> above lists and we see that the carbon has gold, and the ?holes? are <br>
> clean with no gold, only they aren?t clean! The colloidal gold does not <br>
> seem to stick to the plastic film because we see the film if we negative <br>
> stain apoferritin on these same grids or we see the films in the big <br>
> scope based on the ctf or the way the ice has blotted or how the films <br>
> have broken away and flipped onto the carbon. It?s rather obvious when <br>
> you also see true holes next to ?filmed? holes.<br>
> <br>
> Lastly, something will work for a subset of grids in a box, but later <br>
> on, grids from the same box (even the same SIDE of the box) don?t get <br>
> cleaned using the same protocol.<br>
> <br>
> All I can conclude is that whatever this plastic is, we should get <br>
> Quantifoil to stop using it because nothing seems to dissolve it and <br>
> it?s probably killing the planet somehow. If not, it?s certainly <br>
> detrimental to my mental health. Or better yet, Quantifoil should do the <br>
> research and either clean them better themselves or at least give us a <br>
> protocol to do so in the lab.<br>
> <br>
> Any tips, tricks, or surefire ways to clean grids would be great. And <br>
> please publish it because nothing pops up on Pubmed when I search for it.<br>
> <br>
> Thanks,<br>
> <br>
> Cindi L. Schwartz<br>
> <br>
> Electron Microscopist<br>
> <br>
> Rocky Mountain Labs/NIAID/NIH<br>
> <br>
> 903 South 4th Street<br>
> <br>
> Hamilton, MT ?59840<br>
> <br>
> 406-363-9228<br>
> <br>
> Cindi.Schwartz@NIH.gov <mailto:<a href="mailto:Cindi.Schwartz@NIH.gov" target="_blank" rel="noreferrer">Cindi.Schwartz@NIH.gov</a>><br>
> <br>
> Description: Macintosh <br>
> HD:Users:schwartzcl:Desktop:large-green-recycle-symbol-hi.png Please <br>
> remember our Earth before printing this email Description: Macintosh <br>
> HD:Users:schwartzcl:Desktop:large-green-recycle-symbol-hi.png<br>
> <br>
> *************************************************************************<br>
> <br>
> The information in this e-mail and any of its attachments is <br>
> confidential and may contain sensitive information. It should not be <br>
> used by anyone who is not the original intended recipient. If you have <br>
> received this e-mail in error please inform the sender and delete it <br>
> from your mailbox or any other storage devices. National Institute of <br>
> Allergy and Infectious Diseases shall not accept liability for any <br>
> statements made that are sender's own and not expressly made on behalf <br>
> of the NIAID by one of its representatives.<br>
> <br>
> *************************************************************************<br>
> <br>
> <br>
> _______________________________________________<br>
> 3dem mailing list<br>
> <a href="mailto:3dem@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem@ncmir.ucsd.edu</a><br>
> <a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" rel="noreferrer noreferrer" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><br>
> <br>
<br>
-- <br>
Lars-Anders Carlson<br>
Assistant Professor<br>
Dept of Medical Biochemistry and Biophysics<br>
Wallenberg Centre for Molecular Medicine<br>
Ume? University<br>
901 87 Ume?, Sweden<br>
<a href="http://www.carlsonlab.se/" rel="noreferrer noreferrer" target="_blank">http://www.carlsonlab.se/</a><br>
<br>
<br>
------------------------------<br>
<br>
Message: 2<br>
Date: Mon, 1 Jul 2019 17:37:35 +0000<br>
From: Mariena Silvestry Ramos <<a href="mailto:ms3289@cornell.edu" target="_blank" rel="noreferrer">ms3289@cornell.edu</a>><br>
To: "<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem@ncmir.ucsd.edu</a>" <<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem@ncmir.ucsd.edu</a>><br>
Subject: Re: [3dem] Help! Plastic films on my holey grids are KILLING<br>
ME<br>
Message-ID:<br>
<<a href="mailto:BN3PR04MB2260CA5C45471010C90F810ECBF90@BN3PR04MB2260.namprd04.prod.outlook.com" target="_blank" rel="noreferrer">BN3PR04MB2260CA5C45471010C90F810ECBF90@BN3PR04MB2260.namprd04.prod.outlook.com</a>><br>
<br>
Content-Type: text/plain; charset="windows-1252"<br>
<br>
Hi Cindi,<br>
<br>
<br>
I'm so sorry. Years ago, in Phoebe's lab, we used to get the grids with just plastic, then C-coat them ourselves, then dissolve the plastic. And indeed, that plastic was a nightmare to remove.<br>
<br>
<br>
A few yrs ago I shared this email with the list: <a href="https://mail.ncmir.ucsd.edu/pipermail/3dem/2012-December/002760.html" rel="noreferrer noreferrer" target="_blank">https://mail.ncmir.ucsd.edu/pipermail/3dem/2012-December/002760.html</a><br>
<br>
<br>
>From your email, it looks like everything you're doing is pretty much what we did too, but in our experience (if I remember correctly), the plastic did come off.<br>
<br>
<br>
The only thing I didn't see in your email was baking the grids. In Phoebe's lab at Vanderbilt, we put our grids on top of an aluminum disk, took them to a CM12, cranked up the power and left them overnight inside the microscope (our conditions were, a beam spread slightly inside the 4cm mark, spot size 1, all apertures out). I don't think this is done anymore, but in our case, it seemed to work. Dewight used to also mention how obnoxious and stubborn that plastic was, but I don't remember if he had an extra step to completely get rid of the plastic. We did 2x wash with each of the 4 solvents I mention in that email, and waited until the filter paper was dry before adding the next solvent down the list.<br>
<br>
<br>
Finally, IDK if it would be advisable, but for hydrocarbons left behind (this is on holders), we've used an orange peel cleaner (we use Zep, but any other orange-based compound should work similarly). If all else fails, that stuff should eat through anything nasty.<br>
<br>
<br>
Good luck,<br>
<br>
<br>
Mariena<br>
<br>
________________________________<br>
From: 3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem-bounces@ncmir.ucsd.edu</a>> on behalf of Lars-Anders Carlson <<a href="mailto:lars-anders.carlson@umu.se" target="_blank" rel="noreferrer">lars-anders.carlson@umu.se</a>><br>
Sent: Monday, July 1, 2019 1:23:10 PM<br>
To: <a href="mailto:3dem@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem@ncmir.ucsd.edu</a><br>
Subject: Re: [3dem] Help! Plastic films on my holey grids are KILLING ME<br>
<br>
Hi Cindi,<br>
Can you remove it by just plasma cleaning longer? That could be the<br>
reason why some of us typically don't see this film in the first place<br>
(another reason could of course be that we don't look as closely as you).<br>
<br>
QF grids used to have plastic deposits around the edge of the holes,<br>
which is where they would start bubbling at higher exposures (80-100<br>
e/?2), but I haven't seen much of this lately.<br>
<br>
I hope you find a solution since you seem really frustrated about it.<br>
<br>
Cheers,<br>
Lars<br>
<br>
On 2019-07-01 18:38, Schwartz, Cindi (NIH/NIAID) [E] wrote:<br>
> Hi all,<br>
><br>
> We?ve been suffering with trying to get the plastic films off our<br>
> Quantifoil grids with very inconsistent results. We?ve also had film<br>
> issues on the c-flats, but my understanding is that the Quantifoils use<br>
> a plastic background and the c-flats use a carbon background. Please<br>
> correct me if I?m wrong on that one. We use both copper and gold grids<br>
> with the carbon substrate of various different patterns from R1/4 to<br>
> Multi-A with grids that we purchased 2 years ago to ones we?ve received<br>
> last month.<br>
><br>
> The list of solvents we?ve tried that should dissolve plastic films:<br>
><br>
> Ethyl acetate<br>
><br>
> Chloroform<br>
><br>
> Ethylene dichloride<br>
><br>
> Trichlorobenzene<br>
><br>
> The list of solvents to clean them up a bit:<br>
><br>
> Acetone<br>
><br>
> Ethanol<br>
><br>
> 5% acetic acid<br>
><br>
> We?ve tried sonication (a disaster)<br>
><br>
> Submerging in drops<br>
><br>
> Submerging in vast quantities of liquid<br>
><br>
> With/without little stir bars while the grids are on a metal platform in<br>
> the solvent<br>
><br>
> Time ranges from 15 min to overnight<br>
><br>
> With/without a light glow discharge before cleaning<br>
><br>
> We?ve put 10 nm colloidal gold fiducials on both sides of the grids<br>
> (like you?d use for plastic section tomography?where they stick to<br>
> Formvar just fine) after cleaning with various permutations from the<br>
> above lists and we see that the carbon has gold, and the ?holes? are<br>
> clean with no gold, only they aren?t clean! The colloidal gold does not<br>
> seem to stick to the plastic film because we see the film if we negative<br>
> stain apoferritin on these same grids or we see the films in the big<br>
> scope based on the ctf or the way the ice has blotted or how the films<br>
> have broken away and flipped onto the carbon. It?s rather obvious when<br>
> you also see true holes next to ?filmed? holes.<br>
><br>
> Lastly, something will work for a subset of grids in a box, but later<br>
> on, grids from the same box (even the same SIDE of the box) don?t get<br>
> cleaned using the same protocol.<br>
><br>
> All I can conclude is that whatever this plastic is, we should get<br>
> Quantifoil to stop using it because nothing seems to dissolve it and<br>
> it?s probably killing the planet somehow. If not, it?s certainly<br>
> detrimental to my mental health. Or better yet, Quantifoil should do the<br>
> research and either clean them better themselves or at least give us a<br>
> protocol to do so in the lab.<br>
><br>
> Any tips, tricks, or surefire ways to clean grids would be great. And<br>
> please publish it because nothing pops up on Pubmed when I search for it.<br>
><br>
> Thanks,<br>
><br>
> Cindi L. Schwartz<br>
><br>
> Electron Microscopist<br>
><br>
> Rocky Mountain Labs/NIAID/NIH<br>
><br>
> 903 South 4th Street<br>
><br>
> Hamilton, MT 59840<br>
><br>
> 406-363-9228<br>
><br>
> Cindi.Schwartz@NIH.gov <mailto:<a href="mailto:Cindi.Schwartz@NIH.gov" target="_blank" rel="noreferrer">Cindi.Schwartz@NIH.gov</a>><br>
><br>
> Description: Macintosh<br>
> HD:Users:schwartzcl:Desktop:large-green-recycle-symbol-hi.png Please<br>
> remember our Earth before printing this email Description: Macintosh<br>
> HD:Users:schwartzcl:Desktop:large-green-recycle-symbol-hi.png<br>
><br>
> *************************************************************************<br>
><br>
> The information in this e-mail and any of its attachments is<br>
> confidential and may contain sensitive information. It should not be<br>
> used by anyone who is not the original intended recipient. If you have<br>
> received this e-mail in error please inform the sender and delete it<br>
> from your mailbox or any other storage devices. National Institute of<br>
> Allergy and Infectious Diseases shall not accept liability for any<br>
> statements made that are sender's own and not expressly made on behalf<br>
> of the NIAID by one of its representatives.<br>
><br>
> *************************************************************************<br>
><br>
><br>
> _______________________________________________<br>
> 3dem mailing list<br>
> <a href="mailto:3dem@ncmir.ucsd.edu" target="_blank" rel="noreferrer">3dem@ncmir.ucsd.edu</a><br>
> <a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" rel="noreferrer noreferrer" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><br>
><br>
<br>
--<br>
Lars-Anders Carlson<br>
Assistant Professor<br>
Dept of Medical Biochemistry and Biophysics<br>
Wallenberg Centre for Molecular Medicine<br>
Ume? University<br>
901 87 Ume?, Sweden<br>
<a href="http://www.carlsonlab.se/" rel="noreferrer noreferrer" target="_blank">http://www.carlsonlab.se/</a><br>
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</blockquote></div>