<html>
<head>
<meta http-equiv="Content-Type" content="text/html; charset=Windows-1252">
<style type="text/css" style="display:none;"> P {margin-top:0;margin-bottom:0;} </style>
</head>
<body dir="ltr">
<div style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(0, 0, 0);">
Hi Saumya, </div>
<div style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(0, 0, 0);">
If these spots are absent in thicker ice; the black spots you are seeing are likely denaturation at the air water interface because of thin ice.</div>
<div>
<div style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(0, 0, 0);">
<br>
</div>
<div style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(0, 0, 0);">
best regards,</div>
<div style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(0, 0, 0);">
<br>
</div>
<div id="Signature">
<div style="font-family:Calibri,Arial,Helvetica,sans-serif; font-size:12pt; color:rgb(0,0,0)">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">Richard M. Walsh , Ph.D.</span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">The Harvard Cryo-EM </span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">Center for Structural Biology </span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">Harvard Medical School</span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">250 Longwood Avenue</span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">SGM 102</span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">Boston MA 02115</span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">Phone: (617) 432-9042</span><br style="text-align:start; font-family:Helvetica; font-size:12px">
<a href="mailto:Richard_Walsh@hms.harvard.edu" style="text-align:start; font-family:Helvetica; font-size:12px">Email:Richard_Walsh@hms.harvard.edu</a><br style="text-align:start; font-family:Helvetica; font-size:12px">
<span style="text-align:start; font-family:Helvetica; font-size:12px; display:inline!important">Web:<span> </span></span><a href="http://cryoem.hms.harvard.edu/" style="text-align:start; font-family:Helvetica; font-size:12px">cryoem.hms.harvard.edu</a><br>
</div>
</div>
</div>
<div id="appendonsend"></div>
<hr style="display:inline-block;width:98%" tabindex="-1">
<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 Bajaj Saumya (Dr) <Saumya.Bajaj@ntu.edu.sg><br>
<b>Sent:</b> Friday, March 20, 2020 5:49 AM<br>
<b>To:</b> 3dem@ncmir.ucsd.edu <3dem@ncmir.ucsd.edu><br>
<b>Subject:</b> [3dem] Spots in micrographs</font>
<div> </div>
</div>
<style>
<!--
@font-face
{font-family:"Cambria Math"}
@font-face
{font-family:Calibri}
p.x_MsoNormal, li.x_MsoNormal, div.x_MsoNormal
{margin:0cm;
margin-bottom:.0001pt;
font-size:11.0pt;
font-family:"Calibri",sans-serif}
span.x_EmailStyle17
{font-family:"Calibri",sans-serif;
color:windowtext}
.x_MsoChpDefault
{font-family:"Calibri",sans-serif}
@page WordSection1
{margin:72.0pt 72.0pt 72.0pt 72.0pt}
div.x_WordSection1
{}
-->
</style>
<div lang="EN-SG" link="#0563C1" vlink="#954F72">
<div class="x_WordSection1">
<p class="x_MsoNormal">Dear all,</p>
<p class="x_MsoNormal"> </p>
<p class="x_MsoNormal">We are trying to solve the structure of a membrane protein. We have recently started seeing black spots (~1nm) on our micrographs (image attached). The pattern is not uniform, as there are areas where there are no spots. We tried changing
buffer and detergent stock – that did not help. The grids are commercial graphene oxide girds and our buffer contains 0.03% GDN detergent.
</p>
<p class="x_MsoNormal"> </p>
<p class="x_MsoNormal">Have any of you noticed something similar, or can help throw some light on what it could be?
</p>
<p class="x_MsoNormal"> </p>
<p class="x_MsoNormal">Thanks a lot.</p>
<p class="x_MsoNormal">Best regards,</p>
<p class="x_MsoNormal">Saumya</p>
</div>
<hr>
<p style="font-size:10pt; color:#808080; font-family:'Arial'">CONFIDENTIALITY: This email is intended solely for the person(s) named and may be confidential and/or privileged. If you are not the intended recipient, please delete it, notify us and do not copy,
use, or disclose its contents. <br>
Towards a sustainable earth: Print only when necessary. Thank you. </p>
</div>
</body>
</html>