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<div>Dear Yang,</div>
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<div>Adding small amount of detergent may help you to get thinner and smoother ice coverage, thicker ice normally could help random orientations because less surface tension affects the large size particles. If it does not work for your complex, on the specimen
preparation aspect, you may try to glow-discharge on both sides and to add minimum a mount of protein solution, high humidity, maximum force and very short blotting time, under those conditions, you may minimize the complex movement during blotting. If your
complex could generate any weak magnetic field, dilution may be a possible solution. </div>
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<div>On the data collection side, you can try the method Prof. Panczek suggests.</div>
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<div>Good luck,</div>
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<div>Dan</div>
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<div>Dan Shi</div>
<div>JRC, HHMI</div>
<div>19700 Helix Dr</div>
<div>Ashburn, VA 20147 </div>
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<span style="font-weight:bold">From: </span>3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>> on behalf of Yang Li <<a href="mailto:yanglixtal@gmail.com">yanglixtal@gmail.com</a>><br>
<span style="font-weight:bold">Date: </span>Wednesday, May 17, 2017 at 8:46 AM<br>
<span style="font-weight:bold">To: </span>"<a href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>" <<a href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br>
<span style="font-weight:bold">Subject: </span>[3dem] Overcoming orientation preference issue<br>
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<div dir="ltr">Dear colleagues,
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<div>We have a protein sample that suffers from severe orientation preference, that most of the particles cluster into two distinct orientations. This way we have to collect large amounts of data in order to obtain enough effective particles, which hiders us
from reaching high resolution. We have tried to make thicker ice or adding tiny amount of detergent such as Tween20, but not working very well so far. I wonder if there are any tricks we can try to overcome this orientation preference issue? Thank you in advance
for suggestions!</div>
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<div>Best,</div>
<div>Yang </div>
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