http://confocal-microscopy-list.275.s1.nabble.com/Automated-liquid-immersion-tp7590363.html
Following up the previous discussion about automated water/oil immersion, just some information. Besides the mentioned EMBL solution there is also a general (for all microscope brands) solution available from Marzhauser (
). I have no personal experience with it just has seen it at an exhibition. Looked quite nice.
Subject: Re: Spinning disk or (new) point scanning confocals for live imaging of 3D engineered tissues?
Hi Sylvie - I am also very interested but I don't seem to be able to find the product at
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> Hi Sylvie,
>
> I was not aware of a dispenser for oil or silicone, thanks for the tip
> :-)
>
> How do you avoid spillage?
>
> Sincerely,
>
> Matthieu
>
> -----Original Message-----
> From: Confocal Microscopy List <
[hidden email]> On
> Behalf Of Sylvie Le Guyader
> Sent: Sunday, 05 January, 2020 13:25
> To:
[hidden email]
> Subject: Re: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
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> Hi Mattieu
>
> I am curious to know why it would not be possible to use JOBS with the
> silicon objective. To my knowledge you can use JOBS with any objectives.
> Do you perhaps mean that you cannot image a multiwell plate with an
> oil objective? If this is the case, I can say that it is possible
> since we do it at our facility. You need an oil dispenser for your
> objective. We bought one quite a long time ago from EMBLEM, the
> commercial side of EMBL (
https://embl-em.de/). It works without problem.
>
> Med vänlig hälsning / Best regards
> Sylvie
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Facility Manager
> Karolinska Institutet- Bionut Dpt
> Blickagången 16,
> Room 7362 (lab)/7840 (office)
> 14157 Huddinge, Sweden
> mobile: +46 (0) 73 733 5008
> LCI website
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>
> -----Original Message-----
> From: Confocal Microscopy List <
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> Behalf Of VERMEREN Matthieu
> Sent: Sunday, 5 January, 2020 13:55
> To:
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> Subject: Re: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
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> Hi Francesco,
>
> We have a Nikon A1R on Ti2 on demo, resonant scanning combined with
> Nikon's new silicone lenses (long working distance, better match to
> live
> system) means we get excellent images on thick immunostained samples
> (organoids in matrix, tissue slices, young embryos). An organoid that
> would take ~30 minutes to image on a standard system is done in less
> than 6 minutes and with very little noise... meaning some experiments
> that were going to be too long and too expensive are now considered
> do-able by a fair few post docs and PIs. The advantage is that we can
> also use Nikon's JOB for multiwell system (sadly not with the silicone
> lenses, but water lenses are an option).
>
> I'm hoping to test it on Ca++ imaging or on FRET.
>
> Sincerely,
>
> Matthieu
>
> -----Original Message-----
> From: Confocal Microscopy List <
[hidden email]> On
> Behalf Of Francesco Pasqualini
> Sent: 13 December 2019 17:18
> To:
[hidden email]
> Subject: Spinning disk or (new) point scanning confocals for live
> imaging of 3D engineered tissues?
>
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>
> Hi,
> I will start my ERC-funded lab in Italy in 2020, which means I am
> fortunate to be shopping for a confocal instrument right about now.
>
> Since I do mostly live experiments (more details below) I was going to
> get a spinning disk system. But, I realized that the new scanning
> confocals are also relatively fast and gentle. The question is how
> much (if at all) slower and harsher are they with respect to spinning disks?
>
> More details:
> - I do a lot of live experiments (traction force microscopy and
> voltage/calcium-sensitive dyes) on engineered cells (300x300 um FOV)
> and tissues (600x600 um FOV) in 2D or 3D (<300 um thick) that I
> complement with immunostainings after fixation. Needed acquisition
> rates go from <1 fps to 100s depending on the application.
>
> - Originally, I was oriented towards a spinning disk confocal
> (Yokogawa/SORA, Crest) and was looking at ways to deal with the issues
> of confocality on the 3D tissue (600x600x300 um volumes) case. But, I
> realized all vendors have new point scanning confocal (980+Airyscan2,
> FV3000, A1R) with resonant scanners that acquire full frames/ROIs at
> 10s/100s of fps
>
> - Of course, structured illumination (Elyra-7, N-SIM) and light-sheet
> microscopes (QuVi) are also appealing but relatively untested in my
> applications of interests....
>
> The application specialists from all vendors in my area have been
> great to work with but since my lab is not up and running, yet, I
> can't demo these systems directly. Therefore, I could use help and
> feedback, especially from people who have had related experiences in the recent past.
>
> Thanks,
> Francesco
>
> --
> Francesco S. Pasqualini
> Visiting Professor University of Pavia Associate Harvard University
>
> tel: +39 351-521-7788 (IT)
> tel: +1 617-401-5243 (USA)
> The University of Edinburgh is a charitable body, registered in
> Scotland, with registration number SC005336.
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