<html>
<head>
<meta http-equiv="Content-Type" content="text/html; charset=utf-8">
<style type="text/css" style="display:none;"> P {margin-top:0;margin-bottom:0;} </style>
</head>
<body dir="ltr">
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
Hi Jie,</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
<br>
</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
As Ruben mentioned, there is no problem from the point of view of the cryo-TEMs, but the issue is rather about the effect of organic solvent on your sample and on the image quality. The bottom line is that most organic solvents result in much reduced dose tolerance
compared to vitreous ice.</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
<br>
</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
We have imaged inorganic samples <span style="font-size: 14.6667px;">at LN2 temperature
</span>in frozen DMF, which is even more radiation-sensitive than methanol. Adding water, if miscible, can increase the dose-tolerance of the solvent.</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
<br>
</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
For an excellent study about the dose tolerance of various solvents, see this paper by Zhang et al:
<a href="https://urldefense.com/v3/__https://onlinelibrary.wiley.com/doi/full/10.1002/rpm.20230025__;!!Mih3wA!GcaNfE81R9yrQOQH7KwIWIPJHFkPIA2lTyV5mdAQ8Eu9xqw9OWCo-n-7_qu-l3GTTxiJS3Oyzp7hiE2iP5dVcGUaLZQ$">https://onlinelibrary.wiley.com/doi/full/10.1002/rpm.20230025</a></div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
<br>
</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
Best regards,</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
<br>
</div>
<div class="elementToProof" style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
Matthias</div>
<div id="appendonsend"></div>
<div style="font-family: Aptos, Aptos_EmbeddedFont, Aptos_MSFontService, Calibri, Helvetica, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);">
<br>
</div>
<hr style="display: inline-block; width: 98%;">
<div id="divRplyFwdMsg" dir="ltr"><span style="font-family: Calibri, sans-serif; font-size: 11pt; color: rgb(0, 0, 0);"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Ruben Diaz Avalos <rdiaz@lji.org><br>
<b>Sent:</b> Thursday, May 9, 2024 2:59 AM<br>
<b>To:</b> Jie Zhang <jiezhang518@utexas.edu><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu <3dem@ncmir.ucsd.edu><br>
<b>Subject:</b> Re: [3dem] Use of methanol in cryo-EM samples</span>
<div> </div>
</div>
<div style="direction: ltr; font-family: garamond, "times new roman", serif;">Hi Jie,</div>
<div style="direction: ltr; font-family: garamond, "times new roman", serif;"><br>
</div>
<div style="direction: ltr; font-family: garamond, "times new roman", serif;">I have used up to 20% methanol (and ethanol, acetonitrile, acetone...) in some samples; the amount of solvent in the frozen samples is so minimal that there is no way they can cause
any damage to anything, but I can tell you that the beam-induced movement of these samples is much worse than in a purely aqueous buffer.</div>
<div style="direction: ltr; font-family: garamond, "times new roman", serif;"><br>
</div>
<div style="direction: ltr; font-family: garamond, "times new roman", serif;">Ruben.</div>
<br>
<div style="direction: ltr;">On Wed, May 8, 2024 at 9:44 AM Jie Zhang <<a href="mailto:jiezhang518@utexas.edu" id="OWA1bf02d4b-0c8f-ac6b-97a2-7ed7a5c773fc" class="OWAAutoLink" data-loopstyle="linkonly">jiezhang518@utexas.edu</a>> wrote:</div>
<blockquote style="margin: 0px 0px 0px 0.8ex; padding-left: 1ex; border-left: 1px solid rgb(204, 204, 204);">
Hi everyone,<br>
I am trying to use methanol in my cryo-EM samples (specifically for Glacios and Krios). Is there anyone who has prepared the grids with samples containing more than 10& methanol? I just want to make sure high concentration of methanol is okay for these machines.<br>
Thank you.<br>
<br>
Best regards,<br>
Jie Zhang<br>
<br>
<br>
_______________________________________________<br>
3dem mailing list<br>
<a href="mailto:3dem@ncmir.ucsd.edu" id="OWA6e397453-834c-fcf9-473a-f12f6bedcab1" class="OWAAutoLink" data-loopstyle="linkonly">3dem@ncmir.ucsd.edu</a><br>
<a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" id="OWA6eba849b-5d99-fe38-a678-07a95049c3dc" class="OWAAutoLink" data-auth="NotApplicable" data-loopstyle="linkonly">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><br>
</blockquote>
<br>
<div><br>
</div>
--<br>
<div style="direction: ltr;">**********************************************</div>
<div style="direction: ltr;">Ruben Diaz Avalos,</div>
<div style="direction: ltr;">La Jolla Institute for Immunology,</div>
<div style="direction: ltr;">9420 Athena Circle Dr.,</div>
<div style="direction: ltr;">La Jolla, CA 92037</div>
<div style="direction: ltr;">(858)752-6960</div>
<div style="direction: ltr;">**********************************************</div>
</body>
</html>