<html><head><meta http-equiv="content-type" content="text/html; charset=utf-8"></head><body dir="auto">Oui :)<div><br></div><div>That’s the original but there more recent protocols maybe by Harris, with step by step procedures. </div><div><br></div><div>Though the chemicals are poison and may not be used anymore; found on researchgate:</div><div><br></div><div><div class="amp-rd-e-text amp-rd-e-text--size-m amp-rd-e-text--spacing-s amp-rd-e-text--color-inherit redraft-text" style="line-height:1.3;border:0px;margin:0px 0px 15px;padding:0px"><span style="background-color:rgba(255,255,255,0)">"pentylamine or iso-amylamine" technique developed by Dubochet for the protein-free spreading of DNA was substituted, because these amines are poisons, with two others: by Robley Williams, 1977 ("poly lysine technique", this is mentioned above) and by Keller (''BAC technique", Vollenweider HJ, Sogo JM, Koller T. A routine method for protein-free spreading of double- and single-stranded nucleic acid molecules. Proc Natl Acad Sci U S A. 1975 Jan;72(1):83-7.). Both last methods work well and reproducibly. </span></div><div class="amp-rd-e-text amp-rd-e-text--size-m amp-rd-e-text--spacing-s amp-rd-e-text--color-inherit redraft-text" style="line-height:1.3;border:0px;margin:0px 0px 15px;padding:0px"><span style="background-color:rgba(255,255,255,0)">Good luck.</span></div><br><div id="AppleMailSignature" dir="ltr"><div>Sent from my mobile device</div><div>-----------------------------</div><div>Sacha De Carlo, Ph.D. </div><div>Business Development Manager EM</div><div><a href="http://Dectris.com">Dectris.com</a></div></div><div dir="ltr"><br>On 3 Jul 2019, at 22:23, SCHMUTZ Marc (ICS) <<a href="mailto:marc.schmutz@ics-cnrs.unistra.fr">marc.schmutz@ics-cnrs.unistra.fr</a>> wrote:<br><br></div><blockquote type="cite"><div dir="ltr"><div style="font-family:arial,helvetica,sans-serif;font-size:12pt;color:#000000"><div><br></div><div>You mean that paper Sacha ?</div><div><br></div><div><br></div><div><h1 id="screen-reader-main-title" class="Head u-font-serif u-h2 u-margin-s-ver" style="box-sizing:border-box;margin-right:0px;margin-left:0px;padding:0px;color:rgb(80,80,80);word-break:break-word;margin-top:16px!important;margin-bottom:16px!important;font-size:1.5rem!important;font-weight:400!important;line-height:1.333!important;font-family:NexusSerif,Georgia,'Times New Roman',Times,STIXGeneral,'Cambria Math','Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol','Arial Unicode MS',serif!important"><span style="font-family:arial,helvetica,sans-serif;font-size:12pt"><span class="title-text" style="box-sizing:border-box;margin:0px;padding:0px">A new preparation method for dark-field electron microscopy of biomacromolecules</span><a name="baep-article-footnote-id1" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#aep-article-footnote-id1" class="workspace-trigger label" style="box-sizing:border-box;margin:0px 0px 0px 8px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word">1</a><a name="baep-article-footnote-id2" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#aep-article-footnote-id2" class="workspace-trigger label" style="box-sizing:border-box;margin:0px 0px 0px 8px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word">2</a><a name="baep-article-footnote-id3" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#aep-article-footnote-id3" class="workspace-trigger label" style="box-sizing:border-box;margin:0px 0px 0px 8px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word">3</a></span></h1><div class="Banner" id="banner" style="box-sizing:border-box;margin:0px 0px 8px;padding:0px;color:rgb(46,46,46);font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;font-size:14px"><div class="wrapper truncated" style="box-sizing:border-box;margin:0px;padding:0px"><div class="AuthorGroups text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:0.7rem;line-height:1.57"><div class="author-group" id="author-group" style="box-sizing:border-box;margin:0px;padding:0px"><span style="font-family:arial,helvetica,sans-serif;font-size:12pt"><span class="sr-only" style="box-sizing:border-box;padding:0px;border:0px;height:1px;overflow:hidden;width:1px">Author links open overlay panel</span><a class="author size-m workspace-trigger" name="baep-author-id4" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#!" style="box-sizing:border-box;margin:0px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="text given-name" style="box-sizing:border-box;margin:0px;padding:0px">Jacques</span><span class="text surname" style="box-sizing:border-box;margin:0px;padding:0px">Dubochet</span><span class="author-ref" id="bfn1" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline">4</span></span></span></a><a class="author size-m workspace-trigger" name="baep-author-id5" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#!" style="box-sizing:border-box;margin:0px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="text given-name" style="box-sizing:border-box;margin:0px;padding:0px">Michel</span><span class="text surname" style="box-sizing:border-box;margin:0px;padding:0px">Ducommun</span><span class="author-ref" id="bfn1" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline">4</span></span></span></a><a class="author size-m workspace-trigger" name="baep-author-id6" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#!" style="box-sizing:border-box;margin:0px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="text given-name" style="box-sizing:border-box;margin:0px;padding:0px">Max</span><span class="text surname" style="box-sizing:border-box;margin:0px;padding:0px">Zollinger</span></span></a><a class="author size-m workspace-trigger" name="baep-author-id7" href="https://www.sciencedirect.com/science/article/pii/S002253207180148X#!" style="box-sizing:border-box;margin:0px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="text given-name" style="box-sizing:border-box;margin:0px;padding:0px">Edouard</span><span class="text surname" style="box-sizing:border-box;margin:0px;padding:0px">Kellenberger</span><span class="author-ref" id="bfn1" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline">4</span></span></span></a></span></div><div class="author-group" style="box-sizing:border-box;margin:0px;padding:0px"></div><div class="author-group" style="box-sizing:border-box;margin:0px;padding:0px"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><h2 class="publication-title u-h3" style="box-sizing:border-box;margin:0px;padding:0px;color:rgb(80,80,80);font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:center;font-weight:400!important;font-size:1.2rem!important;line-height:1.333!important"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><span style="font-size:12pt;font-family:arial,helvetica,sans-serif"><br></span></span></span></span></span></h2><h2 class="publication-title u-h3" id="publication-title" style="box-sizing:border-box;margin:0px;padding:0px;color:rgb(80,80,80);font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;font-weight:400!important;font-size:1.2rem!important;line-height:1.333!important;text-align:left"><span style="font-size:12pt;font-family:arial,helvetica,sans-serif"><a class="publication-title-link" title="Go to Journal of Ultrastructure Research on ScienceDirect" href="https://www.sciencedirect.com/science/journal/00225320" style="box-sizing:border-box;margin:0px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(80,80,80);word-break:break-word">Journal of Ultrastructure Research</a></span></h2><div class="text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:14px;line-height:1.57;font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:left"><span style="font-size:12pt;font-family:arial,helvetica,sans-serif"><a title="Go to table of contents for this volume/issue" href="https://www.sciencedirect.com/science/journal/00225320/35/1" style="box-sizing:border-box;margin:0px;padding:0px;background-color:transparent;text-decoration:none;color:rgb(12,125,187);word-break:break-word">Volume 35, Issues 1–2</a>, April 1971, Pages 147-167</span></div><div class="text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:14px;line-height:1.57;font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:center"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><br></span></span></span></span></div><div class="text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:14px;line-height:1.57;font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:left">regards </div><div class="text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:14px;line-height:1.57;font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:left"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><br></span></span></span></span></div><div class="text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:14px;line-height:1.57;font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:left"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><br></span></span></span></span></span></span></span></span></div><div class="text-xs" style="box-sizing:border-box;margin:0px;padding:0px;font-size:14px;line-height:1.57;font-family:NexusSans,Arial,Helvetica,'Lucida Sans Unicode','Microsoft Sans Serif','Segoe UI Symbol',STIXGeneral,'Cambria Math','Arial Unicode MS',sans-serif;text-align:left"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline"><span style="font-family:arial,helvetica,sans-serif;font-size:13pt"><span class="content" style="box-sizing:border-box;margin:0px;padding:0px"><span class="author-ref" style="box-sizing:border-box;margin:0px;padding:0px"><span style="box-sizing:border-box;margin:0px;padding:0px;line-height:0;vertical-align:baseline">Marc </span></span></span></span></span></span></span></span></span></span></span></span></div></span></span></span></span></div></div></div></div></div><hr id="zwchr"><div><b>De: </b>"Sacha De Carlo" <<a href="mailto:sacha.decarlo@dectris.com">sacha.decarlo@dectris.com</a>><br><b>À: </b>"Ross, Bradford" <<a href="mailto:bradford.ross@botany.ubc.ca">bradford.ross@botany.ubc.ca</a>><br><b>Cc: </b><a href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a><br><b>Envoyé: </b>Mercredi 3 Juillet 2019 22:09:02<br><b>Objet: </b>Re: [3dem] How to make grids positively charged during or after glow discharge.<br></div><div><br></div><div>You would need to read the old classics by people trying to look at dna in negative staining: look for old protocols using pentylamine (if I find them, I will share them with you) but I’m sure there are some review papers too. <br><div>Best</div><div>Sacha</div><div><br><br><div id="AppleMailSignature" dir="ltr"><div>Sent from my mobile device</div><div>-----------------------------</div><div>Sacha De Carlo, Ph.D. </div><div>Business Development Manager EM</div><div><a href="http://Dectris.com" target="_blank">Dectris.com</a></div></div><div dir="ltr"><br>On 3 Jul 2019, at 20:06, Ross, Bradford <<a href="mailto:bradford.ross@botany.ubc.ca" target="_blank">bradford.ross@botany.ubc.ca</a>> wrote:<br><br></div><blockquote><div dir="ltr">
<div id="divtagdefaultwrapper" style="font-size:12pt;color:#000000;font-family:Calibri,Helvetica,sans-serif" dir="ltr">
<p>Hello Everyone,</p>
<p><br>
</p>
<p>I'm currently trying to image some lipid nanoparticles which have a positive charge on their outer surface. Of course, since normal glow discharge treatment typically yields a negative charge on the grids, the particles are all sticking to the lacey carbon
support instead of remaining suspended in the vitreous ice layer.</p>
<p><br>
</p>
<p>I have heard that Magnesium Acetate can be used to switch the charge from negative to positive after glow discharging, but can't seem to find any protocols for exactly how to treat the grids with it.
<br>
</p>
<p><br>
</p>
<p>Does anyone have a protocol for the use of Magnesium Acetate, or any other suggestions on how to flip the surface charge of the grids to positive? Or other creative solutions I'm not thinking of?<br>
</p>
<p><br>
</p>
<p>Cheers,<br>
</p>
<div id="Signature">
<div style="font-family:Tahoma;font-size:13px">
<div style="font-family:Tahoma;font-size:13px"><font size="2">Bradford Ross<br>
<br>
Electron Microscopy Technician<br>
BioImaging Facility<br>
University of British Columbia<br>
Cunningham Building Rm. 64<br>
2146 East Mall <br>
Vancouver, B.C. <br>
V6T 1Z4<br>
<br>
phone 604-822-6996<br>
</font><br>
</div>
</div>
</div>
</div>
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