<html>
<head>
<meta http-equiv="Content-Type" content="text/html; charset=utf-8">
</head>
<body style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
Hi Linda,
<div class=""><br class="">
</div>
<div class="">Indeed 4% PFA should work but we have also used mixtures of 2%PFA with up to 0.5%GA. When working with pathogenic material we have chosen 2%GA to make sure.</div>
<div class="">GA should in general preserve ultrastructure better but for cryo-CLEM has a lot of autofluorescence.</div>
<div class=""><br class="">
</div>
<div class="">The buffer and not leaving it in the fixative too long (i.e. freeze soon after fixation) are may be even more critical.</div>
<div class=""><br class="">
</div>
<div class="">Best,</div>
<div class="">Paul</div>
<div class=""><br class="">
</div>
<div class="">PS Hope Hugh is doing well<br class="">
<div class="">
<div dir="auto" style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
<div dir="auto" style="caret-color: rgb(0, 0, 0); color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
<div>__________________________________<br class="">
Paul Verkade PhD FRMS<br class="">
Professor of Bioimaging<br class="">
School of Biochemistry<br class="">
Biomedical Sciences Building<br class="">
University of Bristol<br class="">
University Walk<br class="">
BS8 1TD Bristol<br class="">
United Kingdom<br class="">
<br class="">
tel.+44-117-3312179<br class="">
Fax. +44-117-3312168<br class="">
mail: <a href="mailto:p.verkade@bristol.ac.uk" class="">p.verkade@bristol.ac.uk</a><br class="">
websites:<br class="">
<a href="https://urldefense.com/v3/__http://www.bristol.ac.uk/biochemistry/research/pv.html__;!!Mih3wA!QcvpNSzr-s44uHUK_5DdT9C-R-GmWsVGO0dY10_jlcXbCEPJgZSwk8WeT6N4YfWYJQ$" class="">http://www.bristol.ac.uk/biochemistry/research/pv.html</a><br class="">
<a href="https://urldefense.com/v3/__http://bristol.ac.uk/wolfson-bioimaging/equipment/clem/__;!!Mih3wA!QcvpNSzr-s44uHUK_5DdT9C-R-GmWsVGO0dY10_jlcXbCEPJgZSwk8WeT6N_NFR4_A$">http://bristol.ac.uk/wolfson-bioimaging/equipment/clem/</a><br class="">
<br class="">
A top 10 UK university and 58th in the world (QS Rankings 2021)<br class="">
Fourth most targeted UK university by top employers (High Fliers 2020)<br class="">
A top 5 UK university for research (THE analysis of REF 2014)<br class="">
</div>
<div>Ninth best European university for teaching (THE2019)</div>
<div class=""><br class="">
</div>
</div>
<br class="Apple-interchange-newline">
</div>
<br class="Apple-interchange-newline">
<br class="Apple-interchange-newline">
</div>
<div><br class="">
<blockquote type="cite" class="">
<div class="">On 18 Jan 2021, at 11:18, Linda Sandblad <<a href="mailto:linda.sandblad@umu.se" class="">linda.sandblad@umu.se</a>> wrote:</div>
<br class="Apple-interchange-newline">
<div class="">
<div style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
Dear best colleagues!
<div class=""><br class="">
</div>
<div class="">
<div class="">What chemical fixation protocols works for your projects before plunge freezing (or HPF) or what typs of chemical fixation was not good (destroyed more than it helped) prior to cryo-EM sample preparation?</div>
<div class=""><br class="">
</div>
<div class="">Im interested in your experiences, also happy if you have adressed this question in a article and would like to share the link with me.</div>
<div class=""><br class="">
</div>
<div class="">For cryo-EM we prefer to work as close to native as possible, avoiding chemical fixations, but sometimes a fixation is needed to test or move on with tricky projects. For examples, when bacteria cultures are tricky and samples need to be stored,
or when bio safety restricts work in the EM labs (lab specific and different).</div>
<div class=""><br class="">
</div>
<div class="">Best regards, Linda</div>
<div class=""><br class="">
</div>
<div class="">
<div class="">
<div dir="auto" style="caret-color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
<div dir="auto" style="caret-color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
<div dir="auto" style="caret-color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
<div dir="auto" style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">
<div style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;" class="">
Linda Sandblad, researcher and facility director<br class="">
Umeå Core Facility for Electron Microscopy, SciLifeLab Cryo-EM, NMI<br class="">
Department of Chemistry, Umeå University, Sweden<br class="">
Tel: +46 709324936</div>
</div>
</div>
</div>
</div>
</div>
<br class="">
</div>
</div>
</div>
_______________________________________________<br class="">
3dem mailing list<br class="">
<a href="mailto:3dem@ncmir.ucsd.edu" class="">3dem@ncmir.ucsd.edu</a><br class="">
https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem<br class="">
</div>
</blockquote>
</div>
<br class="">
</div>
</body>
</html>