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

[Jaime Llodra]

Dear Long,
Maybe this is overkill but could be interesting to know the opinion of a
civil engineer about how to minimise breakages.

Good luck!
Jaime Llodrá

On Sunday, January 5, 2025, Dom Bellini <dbellini at mrc-lmb.cam.ac.uk> wrote:

> Dear Long,
>
> It seems that you strongly proved that your Krios 4 autoloader is causing
> significant stress on your grids.
>
> 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.
>
> I also assume you did the above a number of times with identical results.
>
> At this point the engineer should be convinced to either fix or replace
> your Krios 4 autoloader.
>
> For example, we do not observe cracking of high numbers of lamellae in our
> Krioses (perhaps 5-10% if any).
>
> I hope you can get this nasty problem sorted as soon as possible.
>
> BW,
>
> D
>
>
> On 5 Jan 2025, at 08:40, Long Gui <guilong at smart.org.cn> wrote:
>
> 
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> --
>
> 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/__;!!Mih3wA!F1u6jVHnx3Nr10x7V9vIzm9w025Tuax00oMi9oIgheUYLGArDg0JHcfW3dzQwlcx1Jg7IS4cxBZwD8ydnXqrRVQ$ 
> content/post_1134771.html
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>
>
> <Broken_lamella copy.jpg>
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