<html><body><div style="font-family: arial, helvetica, sans-serif; font-size: 12pt; color: #000000">Hi Tom, <br>Thanks for the message.<br>I did the tests that you suggested to me.<br><br>I runned the autosharpen tutorial data supplied with Phenix and it worked normally. I blurred that map to 300 A**2 and the resulting map was indeed similar to my map. Finally, I applied the Autosharpen to the blurred map and it was applied a B-sharpen of 222.44 A**2. So, everything worked normally.<br><br>I was thinking that may be disturbing the sharpening is that I have a clearly preferred protein orientation in single-particle analysis. In my data set, the side views are more abundant than the rest of views in my 2D classes (and the number of particles in side view 2D classes are much more larger than in the rest of views 2D classes). What do you think?<br><br>Thanks again for the help.<br>All the best,<br>Carlos.<br><br><hr id="zwchr" data-marker=""><div data-marker=""><b>De: </b>"Tom Terwilliger" <tterwilliger@newmexicoconsortium.org><br><b>Para: </b>"Carlos HENRIQUE FERNANDES" <carlos.henrique_fernandes@sorbonne-universite.fr><br><b>Cc: </b>"phenixbb" <phenixbb@phenix-online.org>, "Tom Terwilliger" <tterwilliger@newmexicoconsortium.org><br><b>Enviadas: </b>Sexta-feira, 21 de maio de 2021 22:16:57<br><b>Assunto: </b>Re: [phenixbb] Problems with sharpening in Autosharpen<br></div><br><div data-marker=""><div dir="ltr">Hi Carlos,<br><div>Normally if you apply a B-sharpen of 220 to a 3 Å <span style="color:rgb(0,0,0);font-family:arial,helvetica,sans-serif;font-size:14.6667px"></span>map, the new map should look really different from the original map (no matter whether you do it yourself or autosharpen does it). So best guess is that something is not happening the way you would like it to be.</div><br><div>A suggestion: run the quick autosharpen tutorial data supplied with Phenix. Presumably it will work normally. Then tell autosharpen to blur that map to 300 A**2. This map should look something like your map (does it?). Now apply autosharpen to the blurred map. It should apply a B-sharpen of about 250 or so (does it?). And the new blurred - then - sharpened map should look good.</div><br><div>Let me know if that doesn't work!</div><div>All the best,</div><div>Tom T</div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Fri, May 21, 2021 at 8:50 AM Carlos HENRIQUE FERNANDES <<a href="mailto:carlos.henrique_fernandes@sorbonne-universite.fr" target="_blank">carlos.henrique_fernandes@sorbonne-universite.fr</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex"><div><div style="font-family:arial,helvetica,sans-serif;font-size:12pt;color:rgb(0,0,0)"><div><div><div><div style="font-family:arial,helvetica,sans-serif;font-size:12pt;color:rgb(0,0,0)"><div><span style="font-size:small"><span style="font-size:11pt">Dear all, <br><br>I am facing some problems with sharpening in Autosharpen. <br>My current map is about 3.2 Å resolution, which I obtained after CTF-Refinement and Polishing in Relion.<br>When I use Autosharpen in default parameters, it tells me that are applied an overall Bsharp applied of 220.34 and a final Biso of 87.42; </span></span><span style="font-size:small"><span style="font-size:11pt">but very little sharpening is done; I was unable to observe a typical helices sharpening at this value of resolution. <br>I did some tests setting progressive Bsharp values (240; 260; 280; 300), but I am still unable to visualize a typical helices sharpening.<br>Has anyone had problems like that? Any tips? <br>Thanks in advance,<br>Carlos.</span></span></div></div></div></div></div></div></div>_______________________________________________<br>
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Unsubscribe: <a href="mailto:phenixbb-leave@phenix-online.org" target="_blank">phenixbb-leave@phenix-online.org</a></blockquote></div><br clear="all"><br>-- <br><div dir="ltr" class="gmail_signature"><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div dir="ltr">Thomas C Terwilliger<div>Laboratory Fellow, Los Alamos National Laboratory</div><div>Senior Scientist, New Mexico Consortium</div><div>100 Entrada Dr, Los Alamos, NM 87544</div><div>Email: <a href="mailto:tterwilliger@newmexicoconsortium.org" target="_blank">tterwilliger@newmexicoconsortium.org</a></div><div>Tel: 505-431-0010</div></div></div></div></div></div></div></div></div></div><br></div></div></body></html>