[3dem] Imaging hexagonal ice crystals (aka "contamination") deliberately

[Michael Elbaum]

Hi Thom,


If you don't need the thin vitreous film, you could probably use continuous carbon, skip the plunging, cool the grid, and wave your hand briskly over the loading station to break the smooth flow of nitrogen gas. Humid air would "contaminate" the cold grid and should give you a nice distribution of ice crystals. This would be easiest in the station of a side-entry holder. I'm not sure how you'd word this later in materials and methods, but it ought to work. Take care that the imaging might be tricky because Bragg diffraction will produce ghost images displaced from the original.


regards,

Michael

________________________________
From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Jan Hübinger <jan.huebinger at hotmail.de>
Sent: Wednesday, September 29, 2021 3:55:35 PM
To: t.sharp at lumc.nl; 3dem at ncmir.ucsd.edu
Subject: Re: [3dem] Imaging hexagonal ice crystals (aka "contamination") deliberately

Hi,

some years ago, we have been imaging ice crystals by cryoEM (no SPA) in aqueous media containing different cryoprotective additives after plunge freezing and subsequent defined warming steps.
https://urldefense.proofpoint.com/v2/url?u=https-3A__www.sciencedirect.com_science_article_pii_S0006349515010000-3Fvia-253Dihub&d=DwIFAw&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=2FYDU976gL-0J11lKp2RBCg6x3L85VV5KJ2Jr-YPI-o&s=KAPI-X3Zeg8koILTxHKXPy9Al4b-SiPF39-YfzUiCpw&e= <https://urldefense.proofpoint.com/v2/url?u=https-3A__www.sciencedirect.com_science_article_pii_S0006349515010000-3Fvia-253Dihub&d=DwMFAw&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=fAzVnNGJDWbKgarjeGEb5dGdPIpChcGjogbOpPUHsFk&s=etFK446Ey_ehOhYslRxg7U0lndxJXLfIhkMpzHZhtHk&e=>
Maybe this helps as a starting point.

Best regards,
Jan

----
Dr. Jan Huebinger
Max-Planck-Institute of Molecular Physiology, Dortmund
jan.huebinger at mpi-dortmund.mpg.de
Phone: +49 231-133 2203


________________________________
Von: 3dem <3dem-bounces at ncmir.ucsd.edu> im Auftrag von t.sharp at lumc.nl <t.sharp at lumc.nl>
Gesendet: Dienstag, 28. September 2021 16:32
An: 3dem at ncmir.ucsd.edu <3dem at ncmir.ucsd.edu>
Betreff: [3dem] Imaging hexagonal ice crystals (aka "contamination") deliberately


Hi all,


I'd like to image hexagonal ice crystals. I see them occasionally as contamination, but not reliably. Now I'd like to vitrify small crystals of hexagonal water ice within an amorphous medium and image them using cryoEM in an analogous way to how we do SPA of proteins. Has anyone done something similar before? My initial ideas were to use a different medium (maybe with high salt/glycerol) and then vitrify in a poor cryogen (e.g., liquid nitrogen), but I've not found any literature yet. Any and all tips welcome!


Thanks in advance for your suggestions,


Thom


Dr. Thomas H. Sharp

Assistant Professor

PI: Bio-Nanopatterning

* 0031-(0)-7152-69499

* t.sharp at lumc.nl<mailto:t.sharp at lumc.nl>

Website: https://urldefense.proofpoint.com/v2/url?u=https-3A__ccb.lumc.nl_about-2Dthe-2Dsharp-2Dlab-2D90&d=DwIFAw&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=2FYDU976gL-0J11lKp2RBCg6x3L85VV5KJ2Jr-YPI-o&s=M5ixhSw046nsmkiuoy8Q2wztw1lTv7EsHhtqr5g_G8k&e= <https://urldefense.proofpoint.com/v2/url?u=https-3A__ccb.lumc.nl_about-2Dthe-2Dsharp-2Dlab-2D90&d=DwMFAw&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=-fx6OPS6KXgM3nes4KiZ13FVKhQYEdMmiQedVib53A8&s=88G21hW0ngyUZqBXMsGPsp8GVqRLkOnr0ovncR2Eok8&e=>

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