[3dem] carbon detaching from grids

Xinghong Dai bestdz at gmail.com
Fri Dec 7 14:02:12 PST 2018


Dear Colleagues,
Talking about Quantifoil, has any of you experienced the same thing as we
do that they just simply ignore your request for quote for weeks, even if
you email them several times? Have never had such a bad shopping experience
before.
Thank you!

Best regards,

_____________________________________________________

Xinghong Dai, Ph.D.

Visiting Assistant Professor

Department of Physiology and Biophysics

Case Western Reserve University, School of Medicine

Email: xinghong.dai at case.edu

Office: +1 216-368 5131            Cell: +1 310-447 4330

Courier delivery: 2109 Adelbert Rd, E610 Robbins Building, Cleveland, OH
44106


On Fri, Dec 7, 2018 at 1:23 PM Bowman, Valorie D <vdb at purdue.edu> wrote:

> Folks,
>
>
> I experienced this as well a couple of years ago- even having some grids
> BAD within a box and others ok!  It's especially bad with peeling off &/or
> breakage of the film if you are using 200 mesh grids: all that open space.
>
>
> It seemed to correlate with the color of the carbon side.  If it was black
> = the carbon film was strong, if it was light or nearly invisible, i.e.
> gold or copper colored, ->  broken films everywhere!  I even tried the
> "thick" films. They were just as inconsistent. Some were black and strong,
> some were shiny and weak.
>
>
> To save heart break, I started screening them and only use the black ones
> "as is".  The shiny one I evaporate a thick layer of carbon over the
> supposed C side, then they work fine too.
>
>
> I know, it's a pain to have to re-work something you *bought to save
> time, * but it's better than sending them back and maybe waiting 6 months
> for a replacement!
>
>
> Let me know if this works for you !   :)
>
>
>
> *Cheers,*
>
>
> *Valorie *
>
> *Valorie Bowman*
>
> vdb at purdue.edu
> 765-494-5643
> EM Facility Laboratory Manager/
> Senior Research Electron Microscopist
> Purdue Cryo EM Facility
> Purdue University
> ------------------------------
> *From:* 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Briegel, A. <
> a.briegel at biology.leidenuniv.nl>
> *Sent:* Friday, December 7, 2018 12:55 PM
> *To:* anna.sartori.rupp at gmail.com
> *Cc:* 3dem at ncmir.ucsd.edu
> *Subject:* Re: [3dem] carbon detaching from grids
>
> Dear Cheri and Anna,
> We also had this problem recently with our standard samples (with same
> glow discharger, plunge freezer and same buffers), so it might  indeed be
> related to the new manufacturing process.
> I hope the problem can be solved.
> Best wishes,
> Ariane
>
>
> Sent from my iPhone
>
> On 7 Dec 2018, at 18:26, Anna Sartori Rupp <anna.sartori.rupp at gmail.com>
> wrote:
>
>
> Hi Cheri,
> We recently experienced exactly the same problem with the two latest
> purchased boxes of quantifoil gold R2/2 grids. We used the same glow
> discharge machine (ELMO) and the same settings that we normally use and we
> experienced that when we put the grids in a buffer like PBS or HEPES + 2%
> PFA the carbon was detaching on almost all the grids.
> To see where the problem could come from we used two very old batches of
> quantifoil grids and repeated the same procedure , but in this case the
> carbon remained nicely attached to the grids.
>
> So it could be that your problem might not be due to your plasma cleaner
> but rather to the new way quantifoil is manifacturing their grids.
>
>  I contacted quantifoil and sent them the corresponding pictures, they
> told me that in fact they recently changed their carbon film deposition
> parameters (he talked about different plasma cleaner conditions...) and
> they told me that they would look into it , but I haven t heard back from
> them yet.
> It would be great if you could also contact them to report your problem.
> I hope this helps,
> Ciao
> Anna
>
> -------------------------------------------------
>
> Anna Sartori-Rupp , PhD
> Ultrastructural Bioimaging Unit
> Institut Pasteur,
> 28 rue du Dr Roux
> 75015 Paris, France
>
>
> Il giorno Gio 6 Dic 2018, 9:12 PM HAMPTON, CHERI M CTR USAF AFMC AFRL/RXAS
> <cheri.hampton.ctr at us.af.mil> ha scritto:
>
>> Hi all,
>>
>>
>> I have suddenly begun experiencing the carbon detaching from my QF R2/1
>> grids when submersed in buffer and when applying proteins for blotting and
>> cryo-plunging. I have tried two different lots of grids, and now I am
>> suspicious of the facility's plasma cleaner that I have been using to glow
>> discharge. All settings and timings have remained constant on my part. I
>> plan to seek out another plasma cleaner to do a comparison test, but
>> meanwhile, what kinds of solvents or materials placed in the plasma
>> cleaner/glow discharge unit might linger and cause problems? I'd like to
>> justify having a "clean" one just for TEM :)
>>
>>
>> Thanks for your input,
>>
>> Cheri
>>
>>  ~'~'~'~'~'~'~'~'~'~'~
>> Cheri M. Hampton, PhD
>> Research Microscopist
>> UES, Inc.
>> AFRL/RXAS
>> 2179 12th St, B652/R122
>> Wright-Patterson AFB, OH 45433-7750
>> (937) 255-9081
>> cheri.hampton.ctr at us.af.mil
>>
>>
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