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<p class="MsoNormal"><span style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US">Dear Jean-Marc,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">one option is to wash the grids.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">See for example the discussion in ccpem last April:<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">“ [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform? “<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">(see answers below, e.g. with ethanol or ethylacetate)<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Bruno<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">###########################################################################<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">Bruno P. Klaholz<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">Centre for Integrative Biology<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">Department of Integrated Structural Biology<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">Institute of Genetics and of Molecular and Cellular Biology<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US">IGBMC - UMR 7104 - U 1258<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US">1, rue Laurent Fries<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">BP 10142
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">67404 ILLKIRCH CEDEX<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">FRANCE<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US">Tel. from abroad: 0033.369.48.52.78<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US">Tel. inside France: 03.69.48.52.78<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US"><a href="https://urldefense.com/v3/__http://www.igbmc.fr/Klaholz__;!!Mih3wA!AWNZ1W8tXR0cqZP0Cc6f2fMC7OmOgyss5KO4rU-tYvAY1UeHOJdoFXGK_Nyz0hLgDc4Sauje4UTj4modSNL9$">http://www.igbmc.fr/Klaholz</a>
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US"><a href="https://urldefense.com/v3/__http://www.igbmc.fr/grandesstructures/cbi__;!!Mih3wA!AWNZ1W8tXR0cqZP0Cc6f2fMC7OmOgyss5KO4rU-tYvAY1UeHOJdoFXGK_Nyz0hLgDc4Sauje4UTj4nnVs9D7$">http://www.igbmc.fr/grandesstructures/cbi</a>
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US"><a href="https://urldefense.com/v3/__https://frisbi.eu__;!!Mih3wA!AWNZ1W8tXR0cqZP0Cc6f2fMC7OmOgyss5KO4rU-tYvAY1UeHOJdoFXGK_Nyz0hLgDc4Sauje4UTj4pOX-dRh$">https://frisbi.eu</a>
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;mso-fareast-language:EN-US"><a href="https://urldefense.com/v3/__http://instruct-eric.com__;!!Mih3wA!AWNZ1W8tXR0cqZP0Cc6f2fMC7OmOgyss5KO4rU-tYvAY1UeHOJdoFXGK_Nyz0hLgDc4Sauje4UTj4p8aFEqy$">http://instruct-eric.com</a>
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="font-size:8.0pt;mso-fareast-language:EN-US"><a href="https://urldefense.com/v3/__https://inext-discovery.eu__;!!Mih3wA!AWNZ1W8tXR0cqZP0Cc6f2fMC7OmOgyss5KO4rU-tYvAY1UeHOJdoFXGK_Nyz0hLgDc4Sauje4UTj4mQSJEyi$">https://inext-discovery.eu</a>
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">-----Original Message-----<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">From: Bruno KLAHOLZ
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Sent: lundi 22 avril 2024 22:13<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">To: 'Sonja Welsch' <sonja.welsch@yahoo.de><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Cc: CCPEM@jiscmail.ac.uk<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Subject: RE: [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform?<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Hi Sonja,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">simply put your grids on a filter paper on a glass petri dish, add some milli-q water (just to about cover the grids), tilt the dish to remove excess (grid will hold on the wet filter
paper by capillarity). Next, add ethylacetate (same, just to about cover the grids), that will help dry the grids to get rid of the water faster and tends to make them more hydrophilic. Let evaporate and dry under a hood; that step is similar to removing formvar.
Cover to protect from dust.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Hope it helps.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Bruno<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">-----Original Message-----<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">From: Sonja Welsch <sonja.welsch@yahoo.de>
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Sent: lundi 22 avril 2024 22:01<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">To: Bruno KLAHOLZ <klaholz@igbmc.fr><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Cc: CCPEM@jiscmail.ac.uk<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Subject: Re: [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform?<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Thanks a lot Terje and Bruno,
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">if ethanol after chloroform works that’s indeed a good sign and very useful to know. Regarding ethyl acetate: I never used that so far - and honestly wasn’t even aware it’s a thing.
Would you perhaps be able to point me to a protocol? <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">Sonja<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> On 22 Apr 2024, at 21:20, Bruno KLAHOLZ <klaholz@igbmc.fr> wrote:<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> Hi Sonja,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> and what about water followed by ethylacetate?<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> Bruno<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> -----Original Message-----<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> From: Collaborative Computational Project in Electron cryo-Microscopy <CCPEM@JISCMAIL.AC.UK> On Behalf Of Dokland, Terje<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US">> Sent: lundi 22 avril 2024 20:19<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US">> To: CCPEM@JISCMAIL.AC.UK<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> Subject: Re: [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform?<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> Hi Sonja,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> If I wash the grids at all, I usually use a wash with chloroform followed by a wash in ethanol, so I’m sure ethanol-stabilized chloroform should work just fine.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> Terje<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> On Apr 22, 2024, at 12:13 PM, Sonja Welsch <00001fc33baed1e2-dmarc-request@JISCMAIL.AC.UK> wrote:<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> [You don't often get email from<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> 00001fc33baed1e2-dmarc-request@jiscmail.ac.uk. Learn why this is<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> important at <a href="https://urldefense.com/v3/__https://aka.ms/LearnAboutSenderIdentification__;!!Mih3wA!AWNZ1W8tXR0cqZP0Cc6f2fMC7OmOgyss5KO4rU-tYvAY1UeHOJdoFXGK_Nyz0hLgDc4Sauje4UTj4md3FGu5$">https://aka.ms/LearnAboutSenderIdentification</a> ]<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> Dear Quantifoil-grid users,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> is anyone out there who can share their experience in washing QF grids<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> (any type) for cryoEM SPA experiments with methanol- or<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> ethanol-stabilised chloroform instead of "pure" chloroform?<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> For some reason our lab chemical suppliers don't seem to be able to<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> ship<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>>> 99% chloroform without stabiliser anymore. I don't really think<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> methanol or ethanol should be harmful to grids, but I'm no chemist and<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> wonder if a stabiliser that potentially changes the polarity of a<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> solvent might change grid "behaviour" in the cryoEM SPA sample prep<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> process... Has anyone tested that and is willing to share their<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> findings? (so our local users don't have to)<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> Best regards,<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> Sonja<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> <o:p>
</o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> --<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> Sonja Welsch<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-GB" style="mso-fareast-language:EN-US">>> Head of Central Electron Microscopy Facility Max Planck Institute of<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US">>> Biophysics Max-von-Laue-Strasse 3<o:p></o:p></span></p>
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<p class="MsoNormal"><b><span lang="EN-US">From:</span></b><span lang="EN-US"> 3dem <3dem-bounces@ncmir.ucsd.edu>
<b>On Behalf Of </b>jean-marc.jeckelmann@unibe.ch<br>
<b>Sent:</b> vendredi 5 juillet 2024 23:32<br>
<b>To:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [3dem] Contaminated Quantifoil® R 1.2/1.3 + 2 nm carbon grids<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.5pt">Dear all,</span><o:p></o:p></p>
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:10.5pt">we have found black-spot-contaminations on several Quantifoil® R 1.2/1.3 + 2 nm carbon grids. The gris originate from four different grid boxes containing
each 100 grids per box (</span>approximate<span style="font-size:10.5pt"> value ~3000 CHF; most severe case see attachment). </span><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:10.5pt">Within the past year we tried several times to reach to Quantifoil and asked for guidance. Unfortunately, Quantifoil never provided us with an answer. </span><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US" style="font-size:10.5pt">Does anybody know how to somehow “clean" these 2 nm carbon layer containing grids ? </span><o:p></o:p></p>
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<p class="MsoNormal">Thanks for your input,<br>
Jean-Marc<o:p></o:p></p>
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<p class="MsoNormal">____________________________<o:p></o:p></p>
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<p class="MsoNormal"><span style="color:black">Jean-Marc Jeckelmann, PhD<br>
Institute of Biochemistry and Molecular Medicine<br>
University of Bern<br>
Bühlstrasse 28<br>
CH-3012 Bern<br>
+41 31 684 3 470<br>
<a href="mailto:jean-marc.jeckelmann@unibe.ch">jean-marc.jeckelmann@unibe.ch</a><o:p></o:p></span></p>
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