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Hi Sacha and <font size="2">Bradford,</font><br>
<br>
This is our version of the pentylamine treatment:<br>
<br>
<a class="moz-txt-link-freetext" href="https://www.ncbi.nlm.nih.gov/pubmed/28580914">https://www.ncbi.nlm.nih.gov/pubmed/28580914</a><br>
<br>
Certainly it should be done with suitable precautions...<br>
<br>
Best wishes<br>
<br>
Paula<br>
<br>
<br>
<div class="moz-cite-prefix">On 03/07/2019 21:33, Sacha De Carlo
wrote:<br>
</div>
<blockquote type="cite"
cite="mid:CAHg0dmzoHdtDncWm65=m28dUcoWEu_FD29Mv0VtRKqEqdSy7+A@mail.gmail.com">
<meta http-equiv="content-type" content="text/html; charset=UTF-8">
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" moz-do-not-send="true">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"
moz-do-not-send="true">marc.schmutz@ics-cnrs.unistra.fr</a>>
wrote:<br>
<br>
</div>
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<div>You mean that paper Sacha ?</div>
<div><br>
</div>
<div><br>
</div>
<div>
<h1 id="screen-reader-main-title" class="Head
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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"
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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"
moz-do-not-send="true">sacha.decarlo@dectris.com</a>><br>
<b>À: </b>"Ross, Bradford" <<a
href="mailto:bradford.ross@botany.ubc.ca"
moz-do-not-send="true">bradford.ross@botany.ubc.ca</a>><br>
<b>Cc: </b><a href="mailto:3dem@ncmir.ucsd.edu"
moz-do-not-send="true">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"
moz-do-not-send="true">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" moz-do-not-send="true">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>
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<pre class="moz-signature" cols="72">--
Paula da Fonseca
MRC Laboratory of Molecular Biology
Cambridge Biomedical Campus
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UK
Tel: + 44 1223 267076
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