<html><head><meta http-equiv="Content-Type" content="text/html; charset=us-ascii"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">Hi Mohamed,<div class=""><br class=""></div><div class="">Have you done CTF refinement, beam tilt in particular?</div><div class=""><br class=""></div><div class="">Basil</div><div class=""><br class=""></div><div class=""><br class=""><div><br class=""><blockquote type="cite" class=""><div class="">Am 03.05.2020 um 07:37 schrieb Mohamed A Sayed <<a href="mailto:mohamed.sobhy@kaust.edu.sa" class="">mohamed.sobhy@kaust.edu.sa</a>>:</div><br class="Apple-interchange-newline"><div class=""><div style="margin-top: 0px; margin-bottom: 0px; caret-color: rgb(0, 0, 0); font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 16px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration: none;" class=""><span style="font-size: 12pt;" class="">Hello,</span></div><div dir="ltr" style="caret-color: rgb(0, 0, 0); font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); text-decoration: none;" class=""><div class=""><div style="margin-top: 0px; margin-bottom: 0px;" class=""><br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">I hope all is safe. If you would please advise on the following:</div><div style="margin-top: 0px; margin-bottom: 0px;" class=""> <br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">I made auto-refinement for a structure u<span style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 16px; background-color: rgb(255, 255, 255);" class="">sing auto-sampling settings: I</span><span style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 16px; background-color: rgb(255, 255, 255);" class="">nitial angular sampling</span><span style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 16px; background-color: rgb(255, 255, 255);" class=""> 3.</span><span style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 16px; background-color: rgb(255, 255, 255);" class="">7, local searching from auto-sampling 1.8. T</span>he output after convergence<span style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 16px; background-color: rgb(255, 255, 255);" class=""> was "Final resolution (without masking) is: 3.26154</span>)". When I generate a mask using binarisation threshold from 0.001 to 0.005, where 0.005 gave warning, the FSC curve has a prominent peak in the FSC plot. I tried changing these parameters and tried the output from other refinement job with bigger box but all give this peak.<br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class=""><br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">my questions<br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">1- What caused and how to get rid of this peak?<br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">2- If this peak problem could not be fixed, can I use proceed with such post-process.star file for ctf refinement and polishing?</div><div style="margin-top: 0px; margin-bottom: 0px;" class=""><br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">Thank you very much for your help and time. <br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class=""><br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class=""><span id="cid:856f3e94-8d33-4343-b90c-0da81494cd59"><pastedImage.png></span><br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class=""><br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class="">Last iteration before conversion: <br class=""></div><div style="margin-top: 0px; margin-bottom: 0px;" class=""><br class=""></div><div class=""> CurrentResolution= 3.93402 Angstroms, which requires orientationSampling of at least 2.04545 degrees for a particle of diameter 220 Angstroms</div><div class=""> Oversampling= 0 NrHiddenVariableSamplingPoints= 1305</div><div class=""> OrientationalSampling= 0.46875 NrOrientations= 145</div><div class=""> TranslationalSampling= 0.4605 NrTranslations= 9</div><div class="">=============================<br class=""></div><div class=""> Oversampling= 1 NrHiddenVariableSamplingPoints= 41760</div><div class=""> OrientationalSampling= 0.234375 NrOrientations= 1160</div><div class=""> TranslationalSampling= 0.23025 NrTranslations= 36<br class=""><span style="font-size: 12pt;" class="">E</span><span style="font-size: 12pt;" class="">xpectation iteration 25</span><br class=""></div><div style="margin-top: 0px; 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