shopping: live-sample confocal+super-res

[Reece, Jeff (NIH/NIDDK) [E]-2]

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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]<https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov>

Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
NIH (National Institutes of Health) /
   NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
8 Center Dr, Rm 126
Bethesda, MD
20892-0851

NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS OTHERWISE SPECIFIED

[George McNamara]

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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/10.1101/677344v2

Nathan 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:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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]<https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov>
>
> Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
> NIH (National Institutes of Health) /
>     NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
> 8 Center Dr, Rm 126
> Bethesda, MD
> 20892-0851
>
> NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS OTHERWISE SPECIFIED

[samuel connell]

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*****

Hi Jeff,

*Commercial Reply*

You've chosen a solid group of instruments for this project. I'll drop you
an email off the list as well; we've been working with the SoRa as an early
adopter beta tester for more than two years now with 18 years of experience
in the Yokogawa SDC domain. We recently hosted a day of SoRa webinars a few
weeks ago. If anyone would like to view the one hour recorded version, let
me know and I'll send you a link.

Cheers,
-
Sam

Samuel Connell
Director of Sales

Intelligent Imaging Innovations (3i)
3575 Ringsby Ct, Suite 102
Denver, CO  80216  USA
1-720-437-6926
www.intelligent-imaging.com
[hidden email]


On Sun, Apr 26, 2020 at 12:16 PM Reece, Jeff (NIH/NIDDK) [E] <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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]<
> https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov
> >
>
> Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
> NIH (National Institutes of Health) /
>    NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
> 8 Center Dr, Rm 126
> Bethesda, MD
> 20892-0851
>
> NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS
> OTHERWISE SPECIFIED
>

[PAVAK SHAH]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

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:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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]<
> https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov
> >
>
> Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
> NIH (National Institutes of Health) /
>    NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
> 8 Center Dr, Rm 126
> Bethesda, MD
> 20892-0851
>
> NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS
> OTHERWISE SPECIFIED
>

[Zdenek Svindrych-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

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:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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]<
> https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov
> >
>
> Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
> NIH (National Institutes of Health) /
>    NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
> 8 Center Dr, Rm 126
> Bethesda, MD
> 20892-0851
>
> NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS
> OTHERWISE SPECIFIED
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Csúcs Gábor-3]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Jeff,

Some very short/minor points to be considered:
1) We have both techniques - for me SIM and AiryScan (SoRa) play in a different league in terms of resolution.
2) I believe that in the mean time you can run the vt-iSIM also with Nikon NIS Elements (which for me is a better option than VisiView). Of course this assumes that you go with a Nikon microscope body.
3) You may want to look also at the "Re-scan" technology (represented by http://www.confocal.nl/). This is a very powerful and also economic solution. We are also planning to invest in a SoRa and my idea is to have on the same system also a Re-Scan setup...  If you buy an appropriate laser box (with multiple outputs), this is a relatively small additional cost..

Greetings Gabor

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Reece, Jeff (NIH/NIDDK) [E]
Sent: Sunday, April 26, 2020 9:07 PM
To: [hidden email]
Subject: shopping: live-sample confocal+super-res

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

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]<https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov>

Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC) NIH (National Institutes of Health) /
   NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
8 Center Dr, Rm 126
Bethesda, MD
20892-0851

NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS OTHERWISE SPECIFIED

[Tim Feinstein]

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I'd follow on George's comments about Leica with a couple observations.  STED does seem excellent for fixed cell imaging, but the iterations that I've used had to make trade-offs with live cell imaging due to speed and the potential for photodamage.  Stefan Hell has published some improvements in live cell STED, so I'd be interested to see how that has translated to commercial systems.  

Note that Leica's implementation of spectral detection doesn't work in real time like Nikon or Zeiss, which use 32-PMT arrays, so it's hard to get as creative with live cell multiplexing.  On the other hand Leica's implementation of FLIM in the FALCON and Stellaris is interesting and could supersede the limitations of spectral detection.  Lifetime imaging *REALLY* expands the system's potential applications.  Ammasi Periasamy at UVA has been promoting FLIM for a long time but it's been a pretty arcane technology, and I'm curious to see how it gets adopted now that there's an accessible consumer model.  I don't have any commercial interest, just a general interest in the technology.  A demo that I attended where we did live cell acceptor photobleach FRET looked promising.  

All the best,


T

Timothy Feinstein, Ph.D.
Research Scientist
University of Pittsburgh Department of Developmental Biology


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara
Sent: Sunday, April 26, 2020 3:45 PM
To: [hidden email]
Subject: Re: shopping: live-sample confocal+super-res

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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://nam05.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.nktphotonics.com%2Flasers-fibers%2Fproduct%2Fsuperk-extend-uv-supercontinuum-extension-unit%2F&amp;data=02%7C01%7Ctnf8%40PITT.EDU%7C0f01b4ce8955456b667608d7ea1b431a%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637235275124621732&amp;sdata=cU3aOnJAH92y9NXNur7hAGsm1hue%2B%2BAUXroujUpOcl0%3D&amp;reserved=0
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://nam05.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.biorxiv.org%2Fcontent%2F10.1101%2F677344v2&amp;data=02%7C01%7Ctnf8%40PITT.EDU%7C0f01b4ce8955456b667608d7ea1b431a%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637235275124621732&amp;sdata=EbkR%2FLXS9IqZJsLg%2FdRLt%2F8bLophTr%2BhcGwx68qHlQY%3D&amp;reserved=0

Nathan 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://nam05.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.abberior-instruments.com%2Fproducts%2Fminflux%2F&amp;data=02%7C01%7Ctnf8%40PITT.EDU%7C0f01b4ce8955456b667608d7ea1b431a%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637235275124621732&amp;sdata=MFsir5oLR4VwEzxdnbs912RBa4w%2BwYif3fDNqqsrTWs%3D&amp;reserved=0

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:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
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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]<https://nam05.safelinks.protection.outlook.com/?url=https%3A%2F%2Fmail.nih.gov%2Fowa%2F14.3.174.1%2Fscripts%2Fpremium%2Fredir.aspx%3FC%3Dvorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.%26URL%3Dmailto%253ajeff.reece%2540nih.gov&amp;data=02%7C01%7Ctnf8%40PITT.EDU%7C0f01b4ce8955456b667608d7ea1b431a%7C9ef9f489e0a04eeb87cc3a526112fd0d%7C1%7C0%7C637235275124626722&amp;sdata=3WbZrinKXWs84T57IQdeaRZNvyLAAeneSfROUpjwR44%3D&amp;reserved=0>
>
> Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
> NIH (National Institutes of Health) /
>     NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
> 8 Center Dr, Rm 126
> Bethesda, MD
> 20892-0851
>
> NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS OTHERWISE SPECIFIED

[ICIT]

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*commercial response*
Dear Jeff,

if you have a free epifluorescence microscope you could also have a look at Clarity, the laser-free spinning disc confocal with structured illumination add-on module from Aurox Ltd (http://www.aurox.co.uk/aurox-confocal-microscope-confocals.php). Basically, any microscope will do, inverted, upright, macro, as long as it has a camera port.

-laser-free and extremely sensitive due to almost 100% emission collection
-xyzt, lambda (other modalities being developed continuously)
-very gentle to samples (excellent for live imaging)
-four freely configurable channels
-up to 100 Hz imaging
-excellent sample penetration
-superresolution with srrf down to 25-30 nm
-multiple cameras and illumination options (often already available)
-extremely user and budget friendly
-no service contract necessary (software updates for free, only two moving parts)
-absolutely fabulous support

Cheers,

>m

[James Kerin]

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&nbsp;***** COMMERCIAL POST *****

Dear Jeff,

If you are considering spinning disk configurations with super-resolution capability then another combination to consider is the Crest X-light v3 spinning disk confocal together with the Gataca Live-SR. The v3 of the X-light is a great and cost effective low light confocal imager, couple this with the equally cost-effective Live-SR for the optical modulation to achieve super-resolution imaging. The combo will take you down to around 120nm resolution.
This recommendation is made from an honest belief that you should take a look and give serious consideration, but please note I have to declare a commercial interest as Cairn use the Crest X-light and Gataca LiveSR in our configurations in the UK and Europe.

https://crestopt.com/xlightv3/

https://www.gataca-systems.com/

Martyn (on James' email account)


Dr Martyn Reynolds
Head of Advanced Imaging
Direct: + 44 (0)7795 304090
&nbsp;

&nbsp;
&nbsp;

&nbsp;
&nbsp;
&nbsp;
&nbsp;
&nbsp;


On 04/26/2020, 08:06pm, "Reece, Jeff (NIH/NIDDK) [E]" ([hidden email]) wrote:
*****
 
To join, leave or search the confocal microscopy listserv, go to:
 
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
 
Post images on http://www.imgur.com and include the link in your posting.
 
*****
 

 
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]&lt;https://mail.nih.gov/owa/14.3.174.1/scripts/premium/redir.aspx?C=vorg2hwQ3EG79HF4VARC2_-txi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.&URL=mailto%3ajeff.reece%40nih.gov&gt;
 

 
Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)
 
NIH (National Institutes of Health) /
 
 NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)
 
8 Center Dr, Rm 126
 
Bethesda, MD
 
20892-0851
 

 
NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS OTHERWISE SPECIFIED

[Cammer, Michael-2]

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*****

Has anyone mentioned yet that if anyone needs to do localized photoactivation, bleaching, ablation, with spinning disk or ISIM you need an additional unit like the Bruker miniscanner with additional lasers and matching filter blocks etc?


Laser scanning confocal may be better for this.


But the white light laser for Leica, which otherwise is an incredible confocal, may not be strong enough?


For most live work, ISIM or spinning disk way better.  Exception would be for yeast and bacteria where we like to zoom in and over sample.


Cheers-


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY  10016

[hidden email]<mailto:[hidden email]>  http://nyulmc.org/micros  http://microscopynotes.com/

Voice direct only, no text or messages:  1-914-309-3270 and 1-646-501-0567



________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of James Kerin <[hidden email]>
Sent: Tuesday, April 28, 2020 10:39 AM
To: [hidden email]
Subject: Re: shopping: live-sample confocal+super-res

[EXTERNAL]

*****
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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=dTYmXyUhHNfpUWhYBbjAA5v_cbCAOV0XRjdZP0RO76Q&e=  and include the link in your posting.
*****

&nbsp;***** COMMERCIAL POST *****

Dear Jeff,

If you are considering spinning disk configurations with super-resolution capability then another combination to consider is the Crest X-light v3 spinning disk confocal together with the Gataca Live-SR. The v3 of the X-light is a great and cost effective low light confocal imager, couple this with the equally cost-effective Live-SR for the optical modulation to achieve super-resolution imaging. The combo will take you down to around 120nm resolution.
This recommendation is made from an honest belief that you should take a look and give serious consideration, but please note I have to declare a commercial interest as Cairn use the Crest X-light and Gataca LiveSR in our configurations in the UK and Europe.

https://urldefense.proofpoint.com/v2/url?u=https-3A__crestopt.com_xlightv3_&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=c3vRjzCudBMPGVJI2zA3jz6uv94ANofm-F10pXNVEOs&e=

https://urldefense.proofpoint.com/v2/url?u=https-3A__www.gataca-2Dsystems.com_&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=kaf25E_h6snd0qfol8M6DgwhvSCiE2G3HtBDQ_zkvK0&e=

Martyn (on James' email account)


Dr Martyn Reynolds
Head of Advanced Imaging
Direct: + 44 (0)7795 304090
&nbsp;

&nbsp;
&nbsp;

&nbsp;
&nbsp;
&nbsp;
&nbsp;
&nbsp;


On 04/26/2020, 08:06pm, "Reece, Jeff (NIH/NIDDK) [E]" ([hidden email]) wrote:
*****

To join, leave or search the confocal microscopy listserv, go to:

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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=dTYmXyUhHNfpUWhYBbjAA5v_cbCAOV0XRjdZP0RO76Q&e=  and include the link in your posting.

*****



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]&lt;https://urldefense.proofpoint.com/v2/url?u=https-3A__mail.nih.gov_owa_14.3.174.1_scripts_premium_redir.aspx-3FC-3Dvorg2hwQ3EG79HF4VARC2-5F-2Dtxi1AZNEITAaQhKx2WUBLeDOG3BM2dSsWeRsCBbyhbstXsPzU2G8.-26URL-3Dmailto-253ajeff.reece-2540nih.gov-26gt-3B&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=qH5QjNZue103aUFQe1A_Ud03OABkPuLGTmi5q-mM7_A&e=



Director, Advanced Light Microscopy & Image Analysis Core (ALMIAC)

NIH (National Institutes of Health) /

 NIDDK (National Institute of Diabetes and Digestive and Kidney diseases)

8 Center Dr, Rm 126

Bethesda, MD

20892-0851



NOTE: THIS EMAIL IS CONFIDENTIAL, FOR THE ADDRESSED AUDIENCE ONLY UNLESS OTHERWISE SPECIFIED

[Alison J. North]

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*****

Hi all,
Interesting discussion.  I just want to address a few specific comments regarding the vt-iSIM system that have come up in other people's posts today.  I spent an entire year trying to decide on the configuration of my iSIM, so hopefully some of this will be helpful info.

1) Sadly Gabor, you cannot run the iSIM through Elements if you are in the USA.  Nikon does not support it over here. Don't get me started on this...

2) Pavak is absolutely correct that if you are imaging thick organisms like zebrafish or worms, the Silicone objectives will be your best choice, and that means either Olympus or Nikon.  HOWEVER, Silicone objectives only work well if you have adjusted the correction collar absolutely perfectly - and this is something that is difficult to do manually without experience, or even (on some systems) if you have lots of experience, due to the objective being hard to access on an inverted system with lots of "stuff". I should know, I have often spent up to 2 hours "nudging" the collar of the Si objective on my OMX system until it is OK (due to the lack of eyepieces), and that is a painful process.  I have spent years urging Olympus and Nikon to motorize the correction collars on their Si objectives, but no luck with it yet, even though Nikon HAS done so for the 100x oil objective.  (Please help me push them on this!) Therefore in the end I chose a Leica stand for my iSIM, using the 93x glycerol objective that DOES have a motorized collar.  Although glycerol is not quite as good an RI match as Silicone oil, we feel that a well-adjusted collar on a glycerol objective works better than a sub-optimally adjusted Si objective, and therefore this is perhaps a better choice for a core facility.   Not that this helps if you mainly work at 60x, as Pavak does, since it's only the 93x objective that is motorized.

3) Regarding the need for a separate FRAP/ablation device at additional cost, I am actually pretty excited with my iSIM system set-up - we have a triple fiber output so that a single set of lasers can be used for the iSIM, the Mizar TILT light sheet module, and also a VisiFRAP module.  These are all controlled by the VisiView software. It has taken quite some months to get everything working properly at full speed and triggering of two cameras etc. etc. , and I haven't yet tested the VisiFRAP module as that was about to be set up when they closed down the University, but the software is certainly allowing super-fast dual colour iSIM and Mizar imaging.  I cannot WAIT to get back to the lab and start using it to image some virus...

All the best,
Alison

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From: Confocal Microscopy List <[hidden email]> on behalf of Cammer, Michael <[hidden email]>
Sent: Tuesday, April 28, 2020 3:16 PM
To: [hidden email] <[hidden email]>
Subject: Re: shopping: live-sample confocal+super-res

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Has anyone mentioned yet that if anyone needs to do localized photoactivation, bleaching, ablation, with spinning disk or ISIM you need an additional unit like the Bruker miniscanner with additional lasers and matching filter blocks etc?


Laser scanning confocal may be better for this.


But the white light laser for Leica, which otherwise is an incredible confocal, may not be strong enough?


For most live work, ISIM or spinning disk way better.  Exception would be for yeast and bacteria where we like to zoom in and over sample.


Cheers-


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY  10016

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From: Confocal Microscopy List <[hidden email]> on behalf of James Kerin <[hidden email]>
Sent: Tuesday, April 28, 2020 10:39 AM
To: [hidden email]
Subject: Re: shopping: live-sample confocal+super-res

[EXTERNAL]

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Dear Jeff,

If you are considering spinning disk configurations with super-resolution capability then another combination to consider is the Crest X-light v3 spinning disk confocal together with the Gataca Live-SR. The v3 of the X-light is a great and cost effective low light confocal imager, couple this with the equally cost-effective Live-SR for the optical modulation to achieve super-resolution imaging. The combo will take you down to around 120nm resolution.
This recommendation is made from an honest belief that you should take a look and give serious consideration, but please note I have to declare a commercial interest as Cairn use the Crest X-light and Gataca LiveSR in our configurations in the UK and Europe.

https://urldefense.proofpoint.com/v2/url?u=https-3A__crestopt.com_xlightv3_&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=c3vRjzCudBMPGVJI2zA3jz6uv94ANofm-F10pXNVEOs&e=

https://urldefense.proofpoint.com/v2/url?u=https-3A__www.gataca-2Dsystems.com_&d=DwIFAw&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=VGygx11l5BTnh0tin0WYmhlIVJwDAlMSr74ybuxIzUE&s=kaf25E_h6snd0qfol8M6DgwhvSCiE2G3HtBDQ_zkvK0&e=

Martyn (on James' email account)


Dr Martyn Reynolds
Head of Advanced Imaging
Direct: + 44 (0)7795 304090
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On 04/26/2020, 08:06pm, "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

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