[3dem] 3dem Digest, Vol 204, Issue 11

Fri Aug 9 16:02:17 PDT 2024

Or one could 3D print a wider backing plate in nylon or ABS.

-da

On Fri, Aug 9, 2024 at 4:00 PM Daniel Asarnow <asarnow at msg.ucsf.edu> wrote:

> I've always used the humidity at 100%, and watched closely to make sure
> the paper touches the grid, and then if the contact breaks early I give it
> another touch with the next spot of the paper before plunging. I also moved
> the blot position in ~1 mm so that I get a mixture of good and hard
> contacts (which might still be good!) instead of a mixture of good and
> failed ones (which are definitely bad).
>
> I do think using a craft knife to cut a piece of foam to the right size
> and replacing the metal plate under the paper with that would solve the
> problem. It's the only difference from the Vitrobot in regards to this
> issue, that the paper isn't backed all the way to the outer edge. There's a
> couple of screws that hold the plate in so I think it would be easy to do.
>
> -da
>
>
> On Fri, Aug 9, 2024 at 12:24 PM David Bulkley <davidbulkley at gmail.com>
> wrote:
>
>> It seems like humidity plays poorly with filter paper in the GP2 for most
>> folks. We typically just leave our humidifier off when we are blotting
>> cells. Is this a bad idea? We've been able to get some decent lamella from
>> cells blotted with
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>> It seems like humidity plays poorly with filter paper in the GP2 for most
>> folks.  We typically just leave our humidifier off when we are blotting
>> cells.  Is this a bad idea?  We've been able to get some decent lamella
>> from cells blotted with the chamber humidifier off, but I'm curious if
>> folks have done any proper testing on this to verify if the cells survive,
>> and if they are damaged, what the symptoms are.  It would suck to find out
>> that, while my filter paper has been doing great, all of my cells have been
>> getting trashed.  Curious if anyone has some real data on this.
>>
>> On Fri, Aug 9, 2024 at 12:00 PM <3dem-request at ncmir.ucsd.edu> wrote:
>>
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>>> Today's Topics:
>>>
>>>    1. Re: Inconsistent Blotting with GP2 (Sen, Anindito)
>>>
>>>
>>> ----------------------------------------------------------------------
>>>
>>> Message: 1
>>> Date: Fri, 9 Aug 2024 17:14:26 +0000
>>> From: "Sen, Anindito" <andysen at tamu.edu>
>>> To: Liz Montabana <liz.montabana at czii.org>, "3dem at ncmir.ucsd.edu"
>>>         <3dem at ncmir.ucsd.edu>
>>> Subject: Re: [3dem] Inconsistent Blotting with GP2
>>> Message-ID:
>>>         <
>>> SN6PR11MB3213B188B5CE3D8FBBCACAE0C7BA2 at SN6PR11MB3213.namprd11.prod.outlook.com
>>> >
>>>
>>> Content-Type: text/plain; charset="us-ascii"
>>>
>>> HI Liz,
>>>
>>> Is the chamber humidity too high (the filter papers get wet?).
>>>
>>> Stay safe & Regards
>>>
>>> Anindito Sen. Ph.D
>>>
>>> Microscopy and Imaging Center
>>> Texas A&M University
>>> 301 Old Main Dr
>>> College Station, TX 77843-2257
>>>
>>> Office: ILSB Room 1133
>>> <
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>>> fax: 979-847-8933
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>>> <
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>>>
>>>
>>>
>>>
>>> From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Liz Montabana <
>>> liz.montabana at czii.org>
>>> Date: Wednesday, August 7, 2024 at 6:59 PM
>>> To: 3dem at ncmir.ucsd.edu <3dem at ncmir.ucsd.edu>
>>> Subject: [3dem] Inconsistent Blotting with GP2
>>> Hi All We are seeing a lot of inconsistent blotting with our GP2 plunge
>>> freezer (The longer we blot, the thicker our grids are, among other odd
>>> behavior) - and we wanted to ask others' experience to see if we could
>>> pinpoint any cause or
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>>> Hi All
>>>
>>> We are seeing a lot of inconsistent blotting with our GP2 plunge freezer
>>> (The longer we blot, the thicker our grids are, among other odd behavior) -
>>> and we wanted to ask others' experience to see if we could pinpoint any
>>> cause or feedback we could provide Leica to enhance the performance.
>>>
>>> Please share your trade tips and secrets for wrangling the GP2 - We've
>>> had experience with several other GP2s and none of them shared this
>>> behavior. Our main samples are whole cells grown on grids.
>>>
>>> Thanks
>>> Liz Montabana
>>> Chan Zuckerberg Imaging Institute
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