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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Hi Bradford.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">We apply a negative glow discharge (Pelco Easiglow with air) to the carbon film of the grid, then immediately add 5 uL of 5 mM Magnesium Acetate solution. This
works for samples that won’t stick to a negatively charged film (liposomes) and works well for us. The reference we have is out of someone’s dissertation (thanks John Lemon!) who cites this reference<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt">94. Dubochet, J.; Groom, M.; Mueller-Neuteboom, S.,
<i>The Mounting of Macromolecules for Electron Microscopy with Particular Reference to Surface Phenomena and the Treatment of Support Films by Glow Discharge</i>. 1982.</span><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Good luck,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Tommi<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Tommi A. White, Ph.D.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Assistant Professor of Biochemistry<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Director, Electron Microscopy Core Facility<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">University of Missouri</span><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">W117 Veterinary Medicine Building<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">1600 East Rollins Street<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Columbia, MO 65211<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">573-882-8304<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><a href="mailto:WhiteTo@missouri.edu">WhiteTo@missouri.edu</a><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><a href="http://emc.missouri.edu/">http://emc.missouri.edu</a><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">@MizzouEMC<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> 3dem <3dem-bounces@ncmir.ucsd.edu>
<b>On Behalf Of </b>Ross, Bradford<br>
<b>Sent:</b> Wednesday, July 03, 2019 1:06 PM<br>
<b>To:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [3dem] How to make grids positively charged during or after glow discharge.<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p><span style="font-family:"Calibri",sans-serif;color:black">Hello Everyone,<o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">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.<o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">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. <o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">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?<o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">Cheers,<o:p></o:p></span></p>
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<p class="MsoNormal" style="margin-bottom:12.0pt"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black">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<o:p></o:p></span></p>
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