http://confocal-microscopy-list.275.s1.nabble.com/Spinning-disk-or-new-point-scanning-confocals-for-live-imaging-of-3D-engineered-tissues-tp7590297p7590351.html
imaging experiments turn out. Francesco
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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.
>
Francesco S. Pasqualini