Posted by
Zdenek Svindrych-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/shopping-live-sample-confocal-super-res-tp7590816p7590833.html
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Hi Jeff,
since you've mentioned VT-iSIM, you may want to add RCM Rescan Confocal
from confocal.nl / Axiom Optics. It's also a fairly basic (and budget)
4-channel confocal unit, it'll work better in densely labeled samples (it's
point scanning), but is slower (1 fps or so).
Other point-scanning instruments (e.g. LSM 980) will not be very fast as
well. Abberior quad scanners tend to be even slower, as the galvo mirrors
have to be bigger. On the other hand, some of their STED microscopes have
adaptive optics built in, which would help with thick samples (but AO is
not trivial to use).
As George mentioned, Leica STED machines are great, and they scan faster
than other point-scanning confocals. And with the WLL, HyDs and integrated
FLIM it's a unique machine...
But with live samples we use spinning disks most of the time, so SoRa would
be a clear choice for us (we demoed the Nikon system and it worked well).
Note, that the zoom optics not only decreases the field of view, but also
the emission light throughput through the disk (the pinholes are 3x smaller
in Airy units). The z-resolution is only slightly improved by the smaller
pinholes (I don't actually know how much, I don't know the actual size of
the pinholes) and the optical reassignment does not improve the optical
z-resolution (the same should be true for VT-iSIM and Rescan), but that's
too much of a theory...
Best, zdenek
On Sun, Apr 26, 2020 at 3:16 PM Reece, Jeff (NIH/NIDDK) [E] <
[hidden email]> wrote:
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>
> Dear List,
>
> We are a core facility ready to make a major purchase, seeking advice.
> The system needs to provide fast, live-sample confocal imaging, but also
> super-res in the 100-150nm range (xy). Here is a sampling of the
> applications we are trying to satisfy:
>
> 1. Z-stacks of cultured cells over time, multi-color labeled.
> Super-res and standard confocal.
> 2. Z-stacks and/or time series of live tissue/organisms (e.g. c.
> elegans, oocytes) up to 40 microns deep (at least), multi-color labeled,
> super-res and standard confocal.
> 3. Z-stack, tile and stitch, super-res of fixed samples, e.g. FISH and
> tissue slices (e.g. mouse kidney).
>
>
>
> We narrowed it down to the following instruments:
>
> 1. Nikon W1 SoRa spinning disk
> 2. Olympus W1 SoRa spinning disk ("SpinSR")
> 3. Visitech vt-iSIM (VisiView software seems to be the best choice here
> in the USA?)
> 4. Zeiss LSM 980 AiryScan 2
> 5. Zeiss Elyra 7 Lattice SIM
>
>
> I will send another email for those that are theoretical-minded; for this
> email, I am interested in practical, hands-on impressions.
> For any of you that have compared any of the above systems, I would
> greatly appreciate to hear those impressions, either to the list or
> directly to me.
> Here are some common categories of comparison that may jog your memory
> and/or provide a framework for your response:
>
> 1. Resolution;
> 2. Speed;
> 3. Sensitivity;
> 4. Photobleaching;
> 5. Maintaining focal plane over time (all the vendors do this well
> now?);
> 6. Color-correction from blue to far red, to edge of image field;
> 7. Usability of software - i.e. user-friendliness, appropriate for a
> core facility;
> 8. Functionality-- i.e. range of features; capability to do what you
> need from a workflow/experimental point of view;
> 9. Reliability, robustness of the system;
> 10. Customer support level.
>
> Stay Safe and Healthy,
> Jeff
>
> Jeff Reece
> Ph: +1.301.451.4330
> E:
[hidden email]<
>
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>
> Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
> NIH (National Institutes of Health) /
> NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
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--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth