[3dem] Layer of Solid Ethane on Cryo Plunge Grids

[YY YY]

Dear Dennis and Xiaochen, thank you very much for your advices! Really appreciate your prompt reply!

Cheers,
Yee Yan
FACTS Lab
Nanyang Technological University
----------------------------------------


On Friday, 10 October 2014, 18:02, Dennis Thomas <thomas at biochem.mpg.de> wrote:
Hi

When I go to put my grids into the box I also often have this layer of  
ethane. To free the grid from the tweezers I just put the grid into  
the slot relaesa the tweezers and move them sideways so that  
contacting the edge of the grid against the side of the slot frees the  
grid. sometimes the ethane comes free and floats away, sometimes not.

I usually look at my grids later and find there is no longer any  
ethane present. Although if you plan to look at then immediately then  
use a longer airlock time 40 vs. 30 seconds for example. I do not  
believe there is any way to 'systematically' assess what the effect  
is. We have been doing cryo successfully for a long time so it would  
seem to not be an important issue. There are so many other potential  
problems that one should consider.

Best wishes

Dennis
-- 
Dennis Thomas Ph.D.
Max Planck Institute for Biochemistry
Dept. of Molecular Structural Biology
Am Klopferspitz 18
D-82152 Martinsried Deutschland
phone: +49-89-85782620



Quoting YY YY <rongchigram79 at yahoo.com.sg>:

> Dear All, i would like to hear from you on advices about the layer  
> of solid ethane formed on the cryo-plunge grids. I read on the past  
> discussions that the thin layer of solid ethane can be useful in  
> protecting the vitrified ice on the grid. I am currently using Gatan  
>  Cryoplunge 3. After the grid is plunged and transferred (and if it  
> isn't blotted it to remove the excess ethane), a thin layer of solid  
>  ethane is formed when i transferred it to the grid box submerged in  
>  the LN2. However, quite often, the tweezer got stuck to the grids  
> and when i tried to detact it, the layer of solid ethane got  
> detacted from the grid as well.
>
>
> May i ask if anyone have systematically studied the effect of such  
> detachment and its impact on the quality of the vitrified ice? I  
> wonder if there is tearing action of the solid ethane from the grid  
> and might damage (or pull out the layer of the vitrified ice as  
> well) the sample?
>
> Best Regards,
> Yee Yan, Tay
> FACTS Lab
> Nanyang Technological University
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