[3dem] Broken/Cracked Lamella during loading to Titan Krios

[Long Gui]

Dear Colleagues,
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.
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.
We would greatly appreciate hearing about your experiences:

 * 
How frequently do you observe lamella that are fine in the Aquilos but become damaged in the Krios?

 * 
Do you have any strategies to test loading forces or reduce the rate of lamella breakage?
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.
Best Regards,
Long Gui
Junior Principal Investigator,
Institute of Bio-Architecture and Bio-Interaction,
Shenzhen Medical Academy of Research and Translation (SMART)
Email: guilong at smart.org.cn
Website: https://urldefense.com/v3/__https://smart.org.cn/en/research/investigator/content/post_1134771.html__;!!Mih3wA!E7-528-k6WExFa9ifKq8D4_DPKgQclaIja2Ue38wnNsOp25FcZlFpxsiMtKOnJtmPeVY_WgBPe99e9Sj-Wsi$  <https://urldefense.com/v3/__https://smart.org.cn/en/research/investigator/content/post_1134771.html__;!!Mih3wA!E7-528-k6WExFa9ifKq8D4_DPKgQclaIja2Ue38wnNsOp25FcZlFpxsiMtKOnJtmPeVY_WgBPe99e9Sj-Wsi$  >
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