<html><body><div style="font-family: arial, helvetica, sans-serif; font-size: 12pt; color: #000000"><div><div><div><div style="font-family: arial, helvetica, sans-serif; font-size: 12pt; color: #000000;" data-mce-style="font-family: arial, helvetica, sans-serif; font-size: 12pt; color: #000000;"><div><span style="font-size: small;" data-mce-style="font-size: small;"><span style="font-size: 11pt;" data-mce-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;" data-mce-style="font-size: small;"><span style="font-size: 11pt;" data-mce-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></body></html>