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<p><span style="font-family: Helvetica; font-size: 12px;">Hi Reza,</span></p>
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<div style="font-family: Helvetica; font-size: 12px;">We have done so much trouble shooting to get the beautiful cryo-EM liposomes images. Based on our experience, one thing that affect the images, is having sucrose in the buffer. The buffer should be as much
as possible close to water, otherwise we saw bubbling effect while we were imaging them. </div>
<div style="font-family: Helvetica; font-size: 12px;">Also, we put the liposome in holy c-flat grids without any extra layer of the carbon. That reduced the background significantly and we got amazing images of the liposome. However, as liposomes are carbon
lover, they got stick to the corner of the holes. Therefore, we played with concentration to get as much as possible of them pushed to the holes and seems 4mg/ml was the best concentration for us.</div>
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<div style="font-family: Helvetica; font-size: 12px;">If you would like, I can send you our liposomes papers with materials and methods on cryo-EM part.</div>
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<div style="font-family: Helvetica; font-size: 12px;">Regards,</div>
<div style="font-family: Helvetica; font-size: 12px;">Aida Razi/ Dr. Ortega's lab</div>
<div style="font-family: Helvetica; font-size: 12px;">McGill University</div>
<div style="font-family: Helvetica; font-size: 12px;">Department of Anatomy and Cell Biology </div>
<div style="font-family: Helvetica; font-size: 12px;">Montreal, QC.</div>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" style="font-size:11pt" color="#000000"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Reza Khayat <rkhayat@ccny.cuny.edu><br>
<b>Sent:</b> May 12, 2017 10:30:36 AM<br>
<b>To:</b> Ludtke, Steven J; 3dem<br>
<b>Subject:</b> Re: [3dem] Liposomes in ice</font>
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<p>Hi,</p>
<p><br>
</p>
<p>We are preparing extruded liposomes (200nm). Problem is we're getting very few liposomes in ice. The starting lipid concentration (DOPG:
<a href="https://avantilipids.com/product/840475/">https://avantilipids.com/product/840475/</a>) is 4mg/ml.</p>
<p><br>
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<p>Reza<br>
</p>
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</p>
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Reza Khayat, PhD
<div>Assistant Professor </div>
<div>City College of New York</div>
<div>Department of Chemistry</div>
<div>New York, NY 10031</div>
</div>
</div>
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<div id="divRplyFwdMsg" dir="ltr"><font style="font-size:11pt" color="#000000" face="Calibri, sans-serif"><b>From:</b> Ludtke, Steven J <sludtke@bcm.edu><br>
<b>Sent:</b> Friday, May 12, 2017 10:27 AM<br>
<b>To:</b> Reza Khayat<br>
<b>Subject:</b> Re: [3dem] Liposomes in ice</font>
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<div>You'll have to define exactly what you mean by liposomes. Extruded SUVs are one of our favorite test/training specimens because they are so easy to prepare and freeze. When most people say liposome, they mean something much larger. You'll need to give
some details if you want useful advice. What are the lipid constituents, how large are the liposomes, how are they prepared, etc. Would also be useful to see a representative image or two.
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<div class="">On May 12, 2017, at 9:18 AM, Reza Khayat <<a href="mailto:rkhayat@ccny.cuny.edu" class="">rkhayat@ccny.cuny.edu</a>> wrote:</div>
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Hi,</div>
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We're trying to visualize liposomes in ice but are having some serious difficulty. Can anyone suggest some tricks that may be helpful? Thanks.</div>
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Best wishes,</div>
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Reza<br class="">
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Reza Khayat, PhD
<div class="">Assistant Professor </div>
<div class="">City College of New York</div>
<div class="">Department of Chemistry</div>
<div class="">New York, NY 10031</div>
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Steven Ludtke, Ph.D.</div>
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Co-Director National Center For Macromolecular Imaging <span class="Apple-converted-space"> </span>(<a href="https://urldefense.proofpoint.com/v2/url?u=http-3A__ncmi.bcm.edu&d=DgMFAg&c=4NmamNZG3KTnUCoC6InoLJ6KV1tbVKrkZXHRwtIMGmo&r=1DzJFW0v6TgEhkW1gy_-ke-RbtvS1fzEbD5_hcb9Up0&m=swGrDni_bnbj0SDRv5hphQJtdOkdoQ-0OVRvykOGmk0&s=vMVLCZ2x3qws1sikvDenN7wwVKbc5CZdCoKuHKU36L8&e=" class="">ncmi.bcm.edu</a>)</div>
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Baylor College of Medicine<span class="Apple-converted-space"> </span> <span class="Apple-converted-space"> </span></div>
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