[3dem] Inconsistent Blotting with GP2

[Daniel Asarnow]

I've noticed it's very common for the paper to break contact with the drop
at the end of blotting, and that the paper tends to wrinkle due to the
humidity in the chamber, causing the actual force to vary wildly between
blots. You can try giving the grid a 2nd short blot, as well as stiffening
the paper with a piece of tape or by stacking two of them.

A modification I have been planning on testing is to add a piece of thin
foam cut to a larger size behind the paper, to provide some backing like on
the vitrobot.

Best,
-da

On Thu, Aug 8, 2024 at 2:16 AM Guillaume Gaullier <
guillaume.gaullier at kemi.uu.se> wrote:

> If you are using the blot sensor, and it doesn’t detect correctly, then it
> cancels the blot (after having touched the grid for a bit) but still
> plunges, and the resulting ice is much thicker than the desired result.
> Normally you should get a
> ZjQcmQRYFpfptBannerStart
> This Message Is From an External Sender
> This message came from outside your organization.
>
> ZjQcmQRYFpfptBannerEnd
> If you are using the blot sensor, and it doesn’t detect correctly, then it
> cancels the blot (after having touched the grid for a bit) but still
> plunges, and the resulting ice is much thicker than the desired result.
> Normally you should get a message on the screen if this is the problem, but
> it’s possible that the blot sensor is misbehaving on your unit (the only
> Leica EM GP2 I used around 2017-18 had a very unreliable blot sensor, so I
> am biased from this experience; maybe newer units don’t have this problem
> anymore).
>
> I hope this helps,
>
> Guillaume
>
> Typed on a phone
>
> On 8 Aug 2024, at 07:20, Michael Elbaum <michael.elbaum at weizmann.ac.il>
> wrote:
>
> 
>
> Hi Liz,     something that comes to mind is a fault that maybe the
> blotting paper doesn't turn. You end up blotting with the same wet area.
> (yes, this happened to me and I didn't pay attention till too late) You can
> mark the paper and just run it through the motions without a tweezer, to
> make sure it turns on every cycle. Something to rule out, at least...
>
> regards,
>
> Michael
> ------------------------------
> *From:* 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Liz Montabana <
> liz.montabana at czii.org>
> *Sent:* Thursday, August 8, 2024 2:59:01 AM
> *To:* 3dem at ncmir.ucsd.edu
> *Subject:* [3dem] Inconsistent Blotting with GP2
>
> *Caution: External Sender. Do not click on links or open attachments
> unless you recognize the sender.*
>
> 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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