Dear Long, <div>Maybe this is overkill but could be interesting to know the opinion of a civil engineer about how to minimise breakages.</div><div><br></div><div>Good luck!</div><div>Jaime Llodrá <br><br>On Sunday, January 5, 2025, Dom Bellini <<a href="mailto:dbellini@mrc-lmb.cam.ac.uk">dbellini@mrc-lmb.cam.ac.uk</a>> wrote:<br><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex"><div dir="auto">Dear Long,<div><br></div><div>It seems that you strongly proved that your Krios 4 autoloader is causing significant stress on your grids. </div><div><br></div><div>You loaded/unloaded grids on a different Krios showing that the lamellae were still intact until you did the same to those grids on Krios 4, which completely cracked all of your lamellae. </div><div><br></div><div>I also assume you did the above a number of times with identical results. </div><div><br></div><div>At this point the engineer should be convinced to either fix or replace your Krios 4 autoloader. </div><div><br></div><div>For example, we do not observe cracking of high numbers of lamellae in our Krioses (perhaps 5-10% if any). </div><div><br></div><div>I hope you can get this nasty problem sorted as soon as possible. </div><div><br></div><div>BW,</div><div><br></div><div>D<br><div dir="ltr"><br></div><div dir="ltr"><br><blockquote type="cite">On 5 Jan 2025, at 08:40, Long Gui <<a href="mailto:guilong@smart.org.cn" target="_blank">guilong@smart.org.cn</a>> wrote:<br><br></blockquote></div><blockquote type="cite"><div dir="ltr"><div style="background-color:#ffeb9c;width:100%;padding:2pt;font-size:10pt;line-height:12pt;font-family:'Calibri';color:black;text-align:left;border:1pt solid #9c6500"><span style="color:#9c6500">CAUTION:</span> This email originated from outside of the LMB:<br>
<b>.-3dem-bounces@ncmir.ucsd.edu-<wbr>.</b><br>
Do not click links or open attachments unless you recognize the sender and know the content is safe.<br>If you think this is a phishing email, please forward it to <a href="mailto:phishing@mrc-lmb.cam.ac.uk" target="_blank">phishing@mrc-lmb.cam.ac.uk</a></div><p>
<br>--<br>
</p><div><div style="font-family:Tahoma,Arial,STHeitiSC-Light,SimSun"><div style="clear:both"><p style="margin:0in;letter-spacing:normal;text-align:start;text-indent:0px;word-spacing:0px;white-space:normal;background-color:rgb(255,255,255)"><span style="color:rgb(36,36,36);font-style:normal;font-weight:400;text-transform:none;font-family:helvetica;font-size:14px">Dear Colleagues,</span></p><p style="margin:0in;letter-spacing:normal;text-align:start;text-indent:0px;word-spacing:0px;white-space:normal;background-color:rgb(255,255,255)"><span style="color:rgb(36,36,36);font-style:normal;font-weight:400;text-transform:none;font-family:helvetica;font-size:14px"><br></span></p><div style="clear:both"><span style="font-family:helvetica;font-size:14px">We are seeking your input regarding a recurring issue with broken or cracked lamella in our newly installed Titan Krios G4. Although lamella (200–300 nm thick) appear to be intact under the Aquilos, more than 60% of them show breakage or cracking after transfer into the Krios, making tilt-series data collection difficult. We have attempted multiple solutions—varying milling patterns, sputtering times, and testing different biological specimens—yet the problem persists.</span></div><div style="clear:both"><span style="font-family:helvetica;font-size:14px"><br></span></div><div style="clear:both"><span style="font-family:helvetica;font-size:14px">Additionally, we discovered that simply one round unloading/loading our lamella grids in the Krios Autoloader can break even previously intact lamella (Shown as the attached image). The engineer team insists that the Autoloader is functioning correctly. However, based on our previous experience with Titan Krios instruments, intact lamella from the Aquilos have typically exhibited a low chance of damage during loading.</span></div><div style="clear:both"><span style="font-family:helvetica;font-size:14px"><br></span></div><div style="clear:both"><span style="font-family:helvetica;font-size:14px">We would greatly appreciate hearing about your experiences:</span></div><ul style="padding:0px 0px 0px 24px;margin:2px 0px;list-style-type:disc"><li><div style="clear:both"><span style="font-family:helvetica;font-size:14px">How frequently do you observe lamella that are fine in the Aquilos but become damaged in the Krios?</span></div></li><li><div style="clear:both"><span style="font-family:helvetica;font-size:14px">Do you have any strategies to test loading forces or reduce the rate of lamella breakage?</span><span style="font-family:helvetica"></span></div></li></ul><div style="clear:both"><span style="font-family:helvetica;font-size:14px"><br></span></div><div style="clear:both"><span style="font-family:helvetica;font-size:14px">If you have encountered similar issues or have suggestions on mitigating lamella damage, we would welcome your advice and comments. Thank you for your time, and we look forward to hearing from you.</span></div><div style="clear:both"><span style="font-family:helvetica;font-size:14px"><br></span></div><div><span style="font-family:helvetica;font-size:14px">Best Regards,</span></div><div><span style="font-family:helvetica;font-size:14px"><br></span></div><div><span style="font-family:helvetica;font-size:14px">Long Gui</span></div><p style="margin:0in;letter-spacing:normal;text-align:start;text-indent:0px;word-spacing:0px;white-space:normal;background-color:rgb(255,255,255)"></p><div style="clear:both"><span style="font-size:15px;font-family:Aptos,sans-serif;color:rgb(36,36,36);font-style:normal;font-weight:400;text-transform:none"><br></span></div><p></p><p style="margin:0in;letter-spacing:normal;text-align:start;text-indent:0px;word-spacing:0px;white-space:normal;background-color:rgb(255,255,255)"></p><div style="clear:both"><span style="font-size:15px;font-family:Aptos,sans-serif;color:rgb(36,36,36);font-style:normal;font-weight:400;text-transform:none"></span><div style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14px;font-style:normal;font-weight:400;letter-spacing:normal;text-align:start;text-indent:0px;text-transform:none;white-space:normal;word-spacing:0px;clear:both"><span style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-style:normal;font-weight:400;text-transform:none;font-family:tahoma;font-size:14px">Junior Principal Investigator,</span></div><div style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14px;font-style:normal;font-weight:400;letter-spacing:normal;text-align:start;text-indent:0px;text-transform:none;white-space:normal;word-spacing:0px;clear:both"><span style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-style:normal;font-weight:400;text-transform:none;font-family:tahoma;font-size:14px">Institute of Bio-Architecture and Bio-Interaction,</span></div><div style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14px;font-style:normal;font-weight:400;letter-spacing:normal;text-align:start;text-indent:0px;text-transform:none;white-space:normal;word-spacing:0px;clear:both"><span style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-style:normal;font-weight:400;text-transform:none;font-family:tahoma;font-size:14px">Shenzhen Medical Academy of Research and Translation (SMART)</span></div><div style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14px;font-style:normal;font-weight:400;letter-spacing:normal;text-align:start;text-indent:0px;text-transform:none;white-space:normal;word-spacing:0px;clear:both"><span style="margin:0px;padding:0px;border:0px;outline:0px;font-family:tahoma;font-size:14px"><span style="margin:0px;padding:0px;border:0px;outline:0px;font-size:14px;font-family:tahoma;color:rgb(0,0,0);font-style:normal;font-weight:400;text-transform:none">Email: <a href="mailto:guilong@smart.org.cn" target="_blank">guilong@smart.org.cn</a></span><span style="margin:0px;padding:0px;border:0px;outline:0px;font-size:14px;font-family:tahoma;color:rgb(0,0,0);font-style:normal;font-weight:400;text-transform:none"><br></span></span></div><div style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-family:Tahoma,Arial,STHeiti,SimSun;font-size:14px;font-style:normal;font-weight:400;letter-spacing:normal;text-align:start;text-indent:0px;text-transform:none;white-space:normal;word-spacing:0px;clear:both"><span style="margin:0px;padding:0px;border:0px;outline:0px;color:rgb(0,0,0);font-style:normal;font-weight:400;text-transform:none;font-family:tahoma;font-size:14px">Website: <a href="https://urldefense.com/v3/__https://smart.org.cn/en/research/investigator/content/post_1134771.html__;!!Mih3wA!E7-528-k6WExFa9ifKq8D4_DPKgQclaIja2Ue38wnNsOp25FcZlFpxsiMtKOnJtmPeVY_WgBPe99e9Sj-Wsi$" target="_blank">https://smart.org.cn/<wbr>en/research/investigator/<wbr>content/post_1134771.html</a></span></div><div></div><span style="font-size:15px;font-family:Aptos,sans-serif;color:rgb(36,36,36);font-style:normal;font-weight:400;text-transform:none"><br></span></div><p></p><p style="margin:0in;letter-spacing:normal;text-align:start;text-indent:0px;word-spacing:0px;white-space:normal;background-color:rgb(255,255,255)"><span style="font-size:15px;font-family:Aptos,sans-serif;color:rgb(36,36,36);font-style:normal;font-weight:400;text-transform:none"><br></span></p></div></div></div>
<div><Broken_lamella copy.jpg></div><span>______________________________<wbr>_________________</span><br><span>3dem mailing list</span><br><span><a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a></span><br><span><a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" target="_blank">https://mail.ncmir.ucsd.edu/<wbr>mailman/listinfo/3dem</a></span><br></div></blockquote></div></div></blockquote></div>