Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/shopping-live-sample-confocal-super-res-tp7590816p7590817.html
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Hi Jeff,
I suggest you extend the list to Leica Stellaris confocal STED, full
load of their new POWER HyDs internal, plus as many more POWER HyDs
(some NIR, i.e. GaAs) on X1 port. White light laser (WLL: modest average
power, but pulsed so much higher ... can also add nearby ex wavelength,
ex. 504 nm + 496 nm + 488 nm for AausFP1). Hopefully also 405 nm laser
for BV421 and all the BVs (and SuperBrights ... and QDots, if
ThermoFisher has solved QD-antibody aggregation issues). Probably a
reach, but Leica (since they've been pushing WLL for many years) and
maybe the other microscope companies, should be looking to get back into
the UV with WLL + SuperK EXTEND-UV
https://www.nktphotonics.com/lasers-fibers/product/superk-extend-uv-supercontinuum-extension-unit/
especially for BUV395 and the other BUVs (and I suppose QDots).
***
A lot of live cell experiments are going to involve fluorescent
proteins, so:
AausFP1 green fluorescent protein should finally obsolete EGFP ... 5x
brighter than EGFP, ~2x brighter than mNeonGreen ... and much narrower
excitation and emission spectra (so ideal for buying WLL) ... and for
many uses, could be tandem dimer, so double those. Amino acid sequence
in figure in text, DNA sequences (not human codon optimized) in supplement
https://www.biorxiv.org/content/10.1101/677344v2Nathan spoke at JHU May 2019: has yellow version (so even more value in
WLL) ... also has new generation other colors (but I've not seen further
bioRxiv preprints or publications).
enjoy,
George
p.s. if not your core, some core at NIH could buy an Abberior
Instruments MinFlux ... 2 nm (and 2 color) "real time" precision
localization,
https://www.abberior-instruments.com/products/minflux/hopefully will come down in price in next couple of years, but the
biology in the meantime could be worth it.
On 4/26/2020 3:06 PM, Reece, Jeff (NIH/NIDDK) [E] wrote:
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>
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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:
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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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