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<p>Dear Umar</p>
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<div>One thing that could be worth checking is to image a range of ice thicknesses in your existing cryo grids. If you are just imaging in the thinnest ice it may be that you have excluded your hexamer and are just seeing smaller complexes. In that case the
hexamers may be hiding in thicker ice. </div>
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<div>This has happened to me in the past and I initially mistakenly concluded that large particles seen in negative stain were being replaced by subcomplexes when I went I imaged in cryo. On checking the thicker ice I realised that this was not in fact the
case.</div>
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<div>Best wishes</div>
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<div>Ed Morris<br>
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<div id="divRplyFwdMsg" dir="ltr"><font color="#000000" face="Calibri, sans-serif" style="font-size:11pt"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Umar Farook <umarfarook12@gmail.com><br>
<b>Sent:</b> 04 June 2020 20:24<br>
<b>To:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [3dem] Hexamer dissociate while freezing Cryo-EM grids</font>
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<span style="color:#9C6500; font-weight:bold">CAUTION:</span> This email originated from outside of the ICR. Do not click links or open attachments unless you recognize the sender's email address and know the content is safe.</div>
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<div dir="auto">Dear All,
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<div dir="auto">I would like to ask the community for suggestions on how to overcome the issue of hexamer dissociating while freezing Cryo-EM grids, the negative staining seems to be perfect hexamer at 0.1 mg/ml concentration. But the sample seems to fall apart
at 1.5 mg/ml in Cryo freezing, please advise, thank you.</div>
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<div dir="auto">Best Regards,</div>
<div dir="auto">Umar</div>
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