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

Fri Aug 9 16:00:54 PDT 2024

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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>>    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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>> <
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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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