[3dem] Gatan 910 multi specimen cryo transfer holder-- not for the neophyte

Tomohiro Nishizawa t-2438 at bs.s.u-tokyo.ac.jp
Wed Oct 3 04:19:00 PDT 2018


Dear Valorie,

Thank you so much for the information.
I also asked Gatan, and they say thy are also developing a new 
multi-specimen holder based on the Elsa holder, though I am not sure how 
good it will be.

Again, I appreciate for your kind response.


Tomohiro,


On 2018/10/02 21:03, Bowman, Valorie D wrote:
> Dear Tomohiro,
> 
> 
> Yes, that problem is corrected.  They no longer sell the version I 
> described.
> 
> 
> The only problem with the version they sell now is the potential to 
> accumulate more frost during the time it takes to load 3 grids and the 
> resulting longer wait for it to stabilize in the column prior to 
> imaging.  However, since it's often used for screening freezing 
> conditions rather than data collection, it's well worth it.
> 
> 
> *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 Tomohiro 
> Nishizawa <t-2438 at bs.s.u-tokyo.ac.jp>
> *Sent:* Tuesday, October 2, 2018 4:33 AM
> *Cc:* 3dem at ncmir.ucsd.edu
> *Subject:* Re: [3dem] Gatan 910 multi specimen cryo transfer holder-- 
> not for the neophyte
> Dear all,
> 
> I found previous posts on Gatan multi-specimen cryo holder 910.
> We are now considering purchasing this holder and am curious about the 
> current situation. Were all these problems already fixed?
> Any information is greatly appreciated.
> 
> Best,
> 
> 
> Tomohiro Nishizawa
> University of Tokyo
> 
> 
> 
> 
> On 2017/11/01 0:17, Matthias Wolf wrote:
>> Dear Valorie,
>>
>> Thank you very much for your detailed reply. This is indeed very helpful.
>>
>> But I don’t fully understand the anatomy of he holder - When you talk 
>> about the “cassette”, what exactly do you mean? From the brochure it 
>> looks just like a 626 with 3 wells at the tip of the holder rod, into 
>> which a naked grid is loaded directly and secured by split ring 
>> (presumably a more “springy” version of the ring used in the 626). 
>> This would be consistent with your “old design”. Does the entire tip 
>> of the holder come off?
>>
>> Or are you loading assembled FEI cartridges (ring, grid, C-clip) 
>> unsecured onto the holder and those can fall out?
>> Their height would probably be too thick for locking them down with 
>> the split ring. I remember that silicon nitride grids, which are about 
>> 0.4 micron thick, could barely be clipped in the 626.
>> If so, is there a dedicated version of the 910, which is designed to 
>> take FEI cartridges (the “new design”)?
>>
>> Also, how many hours of working time do you get before refilling the 
>> dewar?
>> Is a movie rate camera required for drift compensation at anything but 
>> low mag?
>>
>> Thanks, Leo, for pointing out a newer model - I will inquire.
>>
>> Best regards,
>>
>>    Matthias
>>
>> On Oct 31, 2017, at 21:37, Bowman, Valorie D <vdb at purdue.edu 
>> <mailto:vdb at purdue.edu>> wrote:
>>
>>> Dear Matthias,
>>>
>>>
>>> We have one.  It is both great and terrifying.  I have used it to 
>>> screen 6 cryo conditions in an afternoon. BUT, it has one awful 
>>> design flaw- it is incredibly easy to drop the cassette into the 
>>> column. If a person is looking at the 3rd grid and turns the grid 
>>> selector the wrong direction, & plop!  Gravity immediately pulls the 
>>> left end of the cassette down off the edge of the stage, the right 
>>> end pivots up off the ball on the end of the rod, and the whole 
>>> thing falls out into the abyss.
>>>
>>>
>>> We have one of the older models, and that design was far superior. 
>>>  Loading was more difficult, but this risk does not exist with the 
>>> grids mounted in wells in the sample rod itself.
>>>
>>>
>>> Both models take 15-20 minutes longer to stabilize in the microscope 
>>> after insertion, and they take longer to cool down, but I count that 
>>> a fair trade off for 3 samples at once.  The split-ring clip rings 
>>> are not too different from the 626 version; it's just easier to 
>>> accidentally push through a grid.
>>>
>>>
>>> By far, the worst thing is the lack of a real "lock" onto 
>>> the cartridges.  This holder was tested by one of our best people, 
>>> and he dropped the cartridge in the scope the very first time he used 
>>> it!  I know it is only a matter of time before it happens again-- and 
>>> the cartridges are not cheap, plus potential scope down time if 
>>> it blocks the beam, pole pieces, etc.
>>>
>>>
>>> It's a great holder, but if you have a wide range of levels of users, 
>>> it might not be one for everyone.
>>>
>>>
>>> Valorie
>>>
>>>
>>> /Valorie Bowman/
>>>
>>> vdb at purdue.edu <mailto:vdb at purdue.edu>
>>> 494-5643
>>> EM Facility Laboratory Manager/
>>> Senior Research Electron Microscopist
>>> Purdue Cryo EM Facility
>>> Purdue University
>>> ------------------------------------------------------------------------
>>> *From:* 3dem <3dem-bounces at ncmir.ucsd.edu 
>>> <mailto:3dem-bounces at ncmir.ucsd.edu>> on behalf of Matthias Wolf 
>>> <matthias.wolf at oist.jp <mailto:matthias.wolf at oist.jp>>
>>> *Sent:* Monday, October 30, 2017 11:00 PM
>>> *To:* 3dem at ncmir.ucsd.edu <mailto:3dem at ncmir.ucsd.edu>
>>> *Subject:* [3dem] Gatan 910 multi specimen cryo transfer holder
>>>
>>> Hi all,
>>>
>>> We are considering the purchase of a Gatan 910 cryo transfer multi 
>>> specimen side entry holder.
>>>
>>> Does anyone have experience with this holder?
>>>
>>> The specs with respect to stability are identical to the 626 (the 
>>> alternative), but I don’t know how it will perform in practice. It 
>>> can take 3 grids.
>>>
>>> The holder will be used for screening of a larger number of cryo 
>>> samples. Cryo tomography is not a requirement. Due to possible 
>>> contamination during unloading, the idea is not to reuse screened 
>>> grids – once plunging conditions are established, a second grid 
>>> frozen under identical conditions on a reproducible plunging bot will 
>>> be used for data collection on a higher end cryo TEM with DED and 
>>> autoloader. We hope to accelerate the cryo screening by loading 3 
>>> grids at once instead of one at a time and to reduce frosting and ice 
>>> contamination with less cryo transfers. This step aims at offloading 
>>> screening from the expensive high end tools and to maximize their 
>>> beam time for productive data collection. The holder will also be 
>>> used at room temperature for negative stained samples before cryo 
>>> screening, both of which will act as triage steps for samples of 
>>> insufficient quality.
>>>
>>>    Matthias
>>>
>>
>>
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