Posted by
PAVAK SHAH on
URL: http://confocal-microscopy-list.275.s1.nabble.com/shopping-live-sample-confocal-super-res-tp7590816p7590820.html
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Hi Jeff,
In my experience with 2 iSIMs and a SoRa demo (on a Nikon body) I've
consistently found the iSIM to produce brighter images at similar
excitation flux on the sample. I haven't had the time or the right samples
during demo to really rigorously compare resolution with the SoRa,
particularly with or without the 3x extra mag required on paper to make
strong claims either way. I've seen Visiview on Alison North's system at
Rockefeller briefly, used metamorph at MSK, and run my current system using
micro-manager. All three are perfectly serviceable depending on your needs.
We image C. elegans primarily, if you'd like to talk at greater length
offline, feel free to get in touch. We routinely do 1 volume / minute
2-color imaging for lineage tracing in embryos, we've done fast z-stacks
for Ca2+ imaging in freely moving adults at up to 6 volumes per second and
can image embryos fast enough to avoid motion blur during twitching.
Keep in mind that neither the SoRa, the iSIM, nor Airyscan is going to get
quite down to 100 nm.
For thicker live tissue samples, anything you put on a Nikon or Olympus
frame is going to give much better results owing to the availability of
silicone oil immersion objectives. The new Olympus x-line objectives are
also very impressive. Our experience with 3d-SIM on the Elyra (non-lattice)
is that performance suffers much much more strongly with depth compared to
iSIM / Airyscan / SoRa.
As always, for live imaging parallel scanning will be faster and gentler.
Something to keep in mind when debating between point and multi-point
scanning.
Best,
Pavak
On Sun, Apr 26, 2020, 12:17 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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