<div dir="ltr">Dear Colleagues, <div>Talking about Quantifoil, has any of you experienced the same thing as we do that they just simply ignore your request for quote for weeks, even if you email them several times? Have never had such a bad shopping experience before. </div><div>Thank you!</div><div><br></div><div>Best regards,</div><div><p class="MsoNormal" style="color:rgb(80,0,80)">_____________________________________________________</p><p class="MsoNormal" style="color:rgb(80,0,80)">Xinghong Dai, Ph.D.</p><p class="MsoNormal" style="color:rgb(80,0,80)">Visiting Assistant Professor</p><p class="MsoNormal" style="color:rgb(80,0,80)">Department of Physiology and Biophysics</p><p class="MsoNormal" style="color:rgb(80,0,80)">Case Western Reserve University, School of Medicine</p><p class="MsoNormal" style="color:rgb(80,0,80)">Email: <a href="mailto:xinghong.dai@case.edu" target="_blank">xinghong.dai@case.edu</a><br></p><p class="MsoNormal" style="color:rgb(80,0,80)">Office: +1 216-368 5131 Cell: +1 310-447 4330 </p><p class="MsoNormal" style="color:rgb(80,0,80)">Courier delivery: 2109 Adelbert Rd, E610 Robbins Building, Cleveland, OH 44106</p></div><div><br></div><br><div class="gmail_quote"><div dir="ltr">On Fri, Dec 7, 2018 at 1:23 PM Bowman, Valorie D <<a href="mailto:vdb@purdue.edu">vdb@purdue.edu</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex">
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<p>Folks,</p>
<p><br>
</p>
<p>I experienced this as well a couple of years ago- even having some grids BAD within a box and others ok! It's especially bad with peeling off &/or breakage of the film if you are using 200 mesh grids: all that open space. </p>
<p><br>
</p>
<p>It seemed to correlate with the color of the carbon side. If it was black = the carbon film was strong, if it was light or nearly invisible, i.e. gold or copper colored, -> broken films everywhere! I even tried the "thick" films. They were just as inconsistent. Some
were black and strong, some were shiny and weak.</p>
<p><br>
</p>
<p>To save heart break, I started screening them and only use the black ones "as is". The shiny one I evaporate a thick layer of carbon over the supposed C side, then they work fine too.</p>
<p><br>
</p>
<p>I know, it's a pain to have to re-work something you <em>bought to save time, </em>
but it's better than sending them back and maybe waiting 6 months for a replacement! </p>
<p><br>
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<p>Let me know if this works for you ! :) <br>
</p>
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<div style="font-family:Tahoma;font-size:13px"><b>Cheers,</b></div>
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<div style="font-family:Tahoma;font-size:13px"><i>Valorie<br>
</i></div>
<div style="font-family:Tahoma;font-size:13px"><i><br>
</i></div>
<div style="font-family:Tahoma;font-size:13px"><i>Valorie Bowman</i><br>
<br>
<a href="mailto:vdb@purdue.edu" target="_blank">vdb@purdue.edu</a><br>
765-494-5643<br>
EM Facility Laboratory Manager/<br>
Senior Research Electron Microscopist<br>
Purdue Cryo EM Facility<br>
Purdue University</div>
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<div id="m_-1666393536808151870divRplyFwdMsg" dir="ltr"><font style="font-size:11pt" face="Calibri, sans-serif" color="#000000"><b>From:</b> 3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu" target="_blank">3dem-bounces@ncmir.ucsd.edu</a>> on behalf of Briegel, A. <<a href="mailto:a.briegel@biology.leidenuniv.nl" target="_blank">a.briegel@biology.leidenuniv.nl</a>><br>
<b>Sent:</b> Friday, December 7, 2018 12:55 PM<br>
<b>To:</b> <a href="mailto:anna.sartori.rupp@gmail.com" target="_blank">anna.sartori.rupp@gmail.com</a><br>
<b>Cc:</b> <a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br>
<b>Subject:</b> Re: [3dem] carbon detaching from grids</font>
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<div dir="ltr">Dear Cheri and Anna,
<div>We also had this problem recently with our standard samples (with same glow discharger, plunge freezer and same buffers), so it might indeed be related to the new manufacturing process. <br>
I hope the problem can be solved.</div>
<div>Best wishes,</div>
<div>Ariane</div>
<div><br>
</div>
<div><br>
<div id="m_-1666393536808151870AppleMailSignature" dir="ltr">Sent from my iPhone</div>
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On 7 Dec 2018, at 18:26, Anna Sartori Rupp <<a href="mailto:anna.sartori.rupp@gmail.com" target="_blank">anna.sartori.rupp@gmail.com</a>> wrote:<br>
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<div dir="auto"><span style="font-family:sans-serif;font-size:12.8px">Hi Cheri,</span>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">We recently experienced exactly the same problem with the two latest purchased boxes of quantifoil gold R2/2 grids. We used the same glow discharge machine (ELMO) and the same settings that we
normally use and we experienced that when we put the grids in a buffer like PBS or HEPES + 2% PFA the carbon was detaching on almost all the grids.</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">To see where the problem could come from we used two very old batches of quantifoil grids and repeated the same procedure , but in this case the carbon remained nicely attached to the grids.</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"><br>
</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">So it could be that your problem might not be due to your plasma cleaner but rather to the new way quantifoil is manifacturing their grids. </div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"><br>
</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"> I contacted quantifoil and sent them the corresponding pictures, they told me that in fact they recently changed their carbon film deposition parameters (he talked about different plasma cleaner
conditions...) and they told me that they would look into it , but I haven t heard back from them yet. </div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">It would be great if you could also contact them to report your problem.</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">I hope this helps,</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">Ciao </div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">Anna</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"><br>
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<div dir="auto" style="font-family:sans-serif;font-size:12.8px">-------------------------------------------------</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"><br>
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<div dir="auto" style="font-family:sans-serif;font-size:12.8px">Anna Sartori-Rupp , PhD</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">Ultrastructural Bioimaging Unit</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px">Institut Pasteur,</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"><span style="font-size:12.8px">28 rue du Dr Roux</span><br>
</div>
<div dir="auto" style="font-family:sans-serif;font-size:12.8px"><span style="font-size:12.8px">75015 </span><span style="font-size:12.8px">Paris, France </span></div>
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<div dir="ltr">Il giorno Gio 6 Dic 2018, 9:12 PM HAMPTON, CHERI M CTR USAF AFMC AFRL/RXAS <<a href="mailto:cheri.hampton.ctr@us.af.mil" rel="noreferrer noreferrer" target="_blank">cheri.hampton.ctr@us.af.mil</a>> ha scritto:<br>
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<p style="margin-top:0;margin-bottom:0">Hi all,</p>
<p style="margin-top:0;margin-bottom:0"><br>
</p>
<p style="margin-top:0;margin-bottom:0">I have suddenly begun experiencing the carbon detaching from my QF R2/1 grids when submersed in buffer and when applying proteins for blotting and cryo-plunging. I have tried two different lots of grids, and now I am
suspicious of the facility's plasma cleaner that I have been using to glow discharge. All settings and timings have remained constant on my part. I plan to seek out another plasma cleaner to do a comparison test, but meanwhile, what kinds of solvents or materials
placed in the plasma cleaner/glow discharge unit might linger and cause problems? I'd like to justify having a "clean" one just for TEM :)<br>
</p>
<p style="margin-top:0;margin-bottom:0"><br>
</p>
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<div id="m_-1666393536808151870m_6989671788397602273m_5355951194350079210m_-3377630549374303045divtagdefaultwrapper" dir="ltr" style="color:rgb(0,0,0);font-family:Calibri,Helvetica,sans-serif,"EmojiFont","Apple Color Emoji","Segoe UI Emoji",NotoColorEmoji,"Segoe UI Symbol","Android Emoji",EmojiSymbols;font-size:12pt">
<p style="margin-top:0px;margin-bottom:0px">Thanks for your input,</p>
<div><br>
</div>
<div>Cheri</div>
<div><br>
~'~'~'~'~'~'~'~'~'~'~</div>
<div>Cheri M. Hampton, PhD</div>
<div>Research Microscopist</div>
<div>UES, Inc.<br>
AFRL/RXAS<br>
2179 12th St, B652/R122<br>
Wright-Patterson AFB, OH 45433-7750<br>
(937) 255-9081<br>
<a href="mailto:cheri.hampton.ctr@us.af.mil" rel="noreferrer noreferrer noreferrer" target="_blank">cheri.hampton.ctr@us.af.mil</a>
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