[3dem] Contaminated Quantifoil® R 1.2/1.3 + 2 nm carbon grids

Mon Jul 8 03:56:47 PDT 2024

Dear Jean-Marc,

one option is to wash the grids.

See for example the discussion in ccpem last April:
“ [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform? “

(see answers below, e.g. with ethanol or ethylacetate)

Best,

Bruno

###########################################################################
Bruno P. Klaholz
Centre for Integrative Biology
Department of Integrated Structural Biology
Institute of Genetics and of Molecular and Cellular Biology
IGBMC - UMR 7104 - U 1258
1, rue Laurent Fries
BP 10142
67404 ILLKIRCH CEDEX
FRANCE
Tel. from abroad: 0033.369.48.52.78
Tel. inside France: 03.69.48.52.78

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-----Original Message-----
From: Bruno KLAHOLZ
Sent: lundi 22 avril 2024 22:13
To: 'Sonja Welsch' <sonja.welsch at yahoo.de>
Cc: CCPEM at jiscmail.ac.uk
Subject: RE: [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform?

Hi Sonja,

simply put your grids on a filter paper on a glass petri dish, add some milli-q water (just to about cover the grids), tilt the dish to remove excess (grid will hold on the wet filter paper by capillarity). Next, add ethylacetate (same, just to about cover the grids), that will help dry the grids to get rid of the water faster and tends to make them more hydrophilic. Let evaporate and dry under a hood; that step is similar to removing formvar. Cover to protect from dust.
Hope it helps.

Best,

Bruno

-----Original Message-----
From: Sonja Welsch <sonja.welsch at yahoo.de>
Sent: lundi 22 avril 2024 22:01
To: Bruno KLAHOLZ <klaholz at igbmc.fr>
Cc: CCPEM at jiscmail.ac.uk
Subject: Re: [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform?

Thanks a lot Terje and Bruno,
if ethanol after chloroform works that’s indeed a good sign and very useful to know. Regarding ethyl acetate: I never used that  so far - and honestly wasn’t even aware it’s a thing. Would you perhaps be able to point me to a protocol?
Best,
Sonja

> On 22 Apr 2024, at 21:20, Bruno KLAHOLZ <klaholz at igbmc.fr> wrote:
>
>
> Hi Sonja,
>
> and what about water followed by ethylacetate?
>
> Best,
>
> Bruno
>
>
>
> -----Original Message-----
> From: Collaborative Computational Project in Electron cryo-Microscopy <CCPEM at JISCMAIL.AC.UK> On Behalf Of Dokland, Terje
> Sent: lundi 22 avril 2024 20:19
> To: CCPEM at JISCMAIL.AC.UK
> Subject: Re: [ccpem] washing Quantifoil grids with MetOH-stabilised chloroform?
>
> Hi Sonja,
> If I wash the grids at all, I usually use a wash with chloroform followed by a wash in ethanol, so I’m sure ethanol-stabilized chloroform should work just fine.
> Terje
>
>
>
>> On Apr 22, 2024, at 12:13 PM, Sonja Welsch <00001fc33baed1e2-dmarc-request at JISCMAIL.AC.UK> wrote:
>>
>> [You don't often get email from
>> 00001fc33baed1e2-dmarc-request at jiscmail.ac.uk. Learn why this is
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>>
>> Dear Quantifoil-grid users,
>>
>> is anyone out there who can share their experience in washing QF grids
>> (any type) for cryoEM SPA experiments with methanol- or
>> ethanol-stabilised chloroform instead of "pure" chloroform?
>>
>> For some reason our lab chemical suppliers don't seem to be able to
>> ship
>>> 99% chloroform without stabiliser anymore. I don't really think
>> methanol or ethanol should be harmful to grids, but I'm no chemist and
>> wonder if a stabiliser that potentially changes the polarity of a
>> solvent might change grid "behaviour" in the cryoEM SPA sample prep
>> process... Has anyone tested that and is willing to share their
>> findings? (so our local users don't have to)
>>
>> Best regards,
>>
>> Sonja
>>
>>
>> --
>> Sonja Welsch
>> Head of Central Electron Microscopy Facility Max Planck Institute of
>> Biophysics Max-von-Laue-Strasse 3
>> 60438 Frankfurt am Main, Germany
>> Email: sonja.welsch at biophys.mpg.de
>> Mobile: +49 157 80625946
>> Phone: + 49 69 6303 3015
>>

From: 3dem <3dem-bounces at ncmir.ucsd.edu> On Behalf Of jean-marc.jeckelmann at unibe.ch
Sent: vendredi 5 juillet 2024 23:32
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] Contaminated Quantifoil® R 1.2/1.3 + 2 nm carbon grids

Dear all,
we have found black-spot-contaminations on several Quantifoil® R 1.2/1.3 + 2 nm carbon grids. The gris originate from four different grid boxes containing each 100 grids per box (approximate value ~3000 CHF; most severe case see attachment).
Within the past year we tried several times to reach to Quantifoil and asked for guidance. Unfortunately, Quantifoil never provided us with an answer.
Does anybody know how to somehow “clean" these 2 nm carbon layer containing grids ?
Thanks for your input,
Jean-Marc

[cid:image001.jpg at 01DAD136.497BCBE0]

____________________________
Jean-Marc Jeckelmann, PhD
Institute of Biochemistry and Molecular Medicine
University of Bern
Bühlstrasse 28
CH-3012 Bern
+41 31 684 3 470
jean-marc.jeckelmann at unibe.ch<mailto:jean-marc.jeckelmann at unibe.ch>

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