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<p>Is the protein layer associated with either (or both) the layers of surface ice contamination? That would be a clue.<br>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" color="#000000" style="font-size:11pt"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Schmid, Michael F. <m-schmid@slac.stanford.edu><br>
<b>Sent:</b> Thursday, August 10, 2017 10:04 PM<br>
<b>To:</b> Sonali Dhindwal; 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> Re: [3dem] Tomography to visualize interaction of liposomes</font>
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<p>Possibly, the protein is attracted to the air-water interface. In bulk biochemical measurements (maybe also with negative stain), this is not a problem; and with single particle methods, you can't detect the problem you are seeing in tomograms, but with
the thin layer after blotting, there is a lot more surface to volume ratio for the protein to be interacting with. Just a guess.<br>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" color="#000000" style="font-size:11pt"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Sonali Dhindwal <sdhindwal@ccny.cuny.edu><br>
<b>Sent:</b> Thursday, August 10, 2017 9:56 PM<br>
<b>To:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [3dem] Tomography to visualize interaction of liposomes</font>
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<div><span style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small; background-color:rgb(255,255,255)">Dear 3DEM group members,</span>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small"><br>
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<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">I have been studying the interaction of my protein of interest (~900 KDa and ~200Å in size) with liposomes (~2000Å is size). </div>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">Biochemical assays demonstrates their interaction and so is the images collected by negative stain and Cryo-EM. </div>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">To clearly visualize interaction, I have collected few tomograms. Though, after processing the tomograms and going through the slices, liposomes and protein seem to be in different
layers.</div>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">In initial slices, I can see all the liposomes and in the later slices I see all the protein molecules. And going through the slices, first I see liposomes and as they start to
fade away, I start to see the protein molecules and few protein molecules lying very close to the liposomes. </div>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">Is this normal ? </div>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">I am looking for any suggestions or advice that can be done either during processing of data or preparation of grids that can help to improve the visualization of interactions
among these molecules.</div>
<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">Thanks and Regards<br>
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<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small"><br>
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<div style="color:rgb(34,34,34); font-family:arial,sans-serif; font-size:small">-- <br>
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<div>Sonali Dhindwal</div>
<div>Post Doc<br>
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"<font color="#444444" size="2"><span style="line-height:16px">The dream is not that you see in sleep, dream is which does not let you sleep</span></font>."<br>
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