Yokogawa W1 spinning disk with OPRA-style superresolution

[Zdenek Svindrych-2]

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Dear listers,

when did they introduce this? I must have missed it...
https://www.visitron.de/products/visiscope-confocal/yokogawa-sora.html
I know Yokogawa engineers published it back in 2015, but I did not know
it's finally available.

Has anyone tried/purchased it? What are the impressions?

On a different note, anyone familiar with the Visitron's beam homogenizing
system for CSU-W1 willing to share some details? How does it compare to
"Andor's" Borealis? The thing is that we're observing some strange
borealis-related effects at short exposures; moreover, Andor does not sell
W1 with borealis anymore...

Thanks!

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

[Emmanuel Levy]

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Hi,

I have no experience with the SORA add-on. One thing to keep in mind is
that it decreases the field of view by 75% (because of the 2x
magnification).

On a different note, anyone familiar with the Visitron's beam homogenizing
> system for CSU-W1 willing to share some details? How does it compare to
> "Andor's" Borealis? The thing is that we're observing some strange
> borealis-related effects at short exposures; moreover, Andor does not sell
> W1 with borealis anymore...
>

We've had the homogenizer for a while. The homogeneity is better than with
regular W1 optics but not flat. In our system we get ~10-15% variation
across the FOV for GFP, 20-25% for RFP, and about or above 30% for BFP.

Hope this helps,

Emmanuel

>
> Thanks!
>
> Best, zdenek
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Associate - Imaging Specialist
> Department of Biochemistry and Cell Biology
> Geisel School of Medicine at Dartmouth
>

[Sylvie Le Guyader]

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The SoRa will be at FOM and we will get an in-house demo on a Nikon Ti2 right after that. The Ti2 has a very large field of view, which I think compensates for the 2x lens.

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7C,
Room 7362 (lab)/7840 (office)
14157 Huddinge, Sweden
mobile: [phone]
LCI website
Follow our microscopy blog!



-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Emmanuel Levy
Sent: den 21 februari 2019 10:43
To: [hidden email]
Subject: Re: Yokogawa W1 spinning disk with OPRA-style superresolution

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*****

Hi,

I have no experience with the SORA add-on. One thing to keep in mind is that it decreases the field of view by 75% (because of the 2x magnification).

On a different note, anyone familiar with the Visitron's beam homogenizing
> system for CSU-W1 willing to share some details? How does it compare
> to "Andor's" Borealis? The thing is that we're observing some strange
> borealis-related effects at short exposures; moreover, Andor does not
> sell
> W1 with borealis anymore...
>

We've had the homogenizer for a while. The homogeneity is better than with regular W1 optics but not flat. In our system we get ~10-15% variation across the FOV for GFP, 20-25% for RFP, and about or above 30% for BFP.

Hope this helps,

Emmanuel

>
> Thanks!
>
> Best, zdenek
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Associate - Imaging Specialist Department of Biochemistry and
> Cell Biology Geisel School of Medicine at Dartmouth
>

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[Ian Dobbie]

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Zdenek Svindrych <[hidden email]> writes:

> *****
> 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 listers,
>
> when did they introduce this? I must have missed it...
> https://www.visitron.de/products/visiscope-confocal/yokogawa-sora.html
> I know Yokogawa engineers published it back in 2015, but I did not know
> it's finally available.
>
> Has anyone tried/purchased it? What are the impressions?

We played with one on an Olympus setup. It is slightly different than the
standard Yokogawa setup as they have their own beam expansion optics and
then the off the shelf SORA disk.

Seems to work pretty well. Definitely getting super-resolution data but
not convinced by the Olympus weiner filtering and deconvolution
idea. Also seems very expensive in the UK, ball park price seems to be
£350-500,000 depending on options.

It is easy to go from conventional spinning disk to the SORA mode, and
the lenses in front of the pinholes mean you get significantly more light
through. The PSF on thick samples can be pretty horrific though. I think
it would be very hard to deconvolve in bright thick samples as the out
of focus bleed through from other excitation spots is very image
position dependant, you can see that arc's due to the pinhole positions
on the disk.

Ian

[Sylvie Le Guyader]

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Hi Ian



I have a couple of questions:

-          Is the price you mention for an upgrade (i.e. the Olympus microscope was yours) or is it for a full system with lasers, camera…?

-          When you talk about? Do you think that the pinhole crosstalk you mention is worse with SoRa than with the conventional/standard Yokogawa spinning disk (by ‘conventional/standard’, do you mean W1 or X1?)? We have an X1 we are very happy with but if we don’t clear the sample and we image thick samples, we get the expected pinhole crosstalk.



Med vänlig hälsning / Best regards



Sylvie



@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD

Live Cell Imaging Facility Manager

Karolinska Institutet- Bionut Dpt

Hälsovägen 7C,

Room 7362 (lab)/7840 (office)

14157 Huddinge, Sweden

mobile: [phone]

LCI website

Follow our microscopy blog!





-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Ian Dobbie
Sent: den 27 februari 2019 12:45
To: [hidden email]
Subject: Re: Yokogawa W1 spinning disk with OPRA-style superresolution



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Zdenek Svindrych <[hidden email]<mailto:[hidden email]>> writes:



> *****

> 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 listers,

>

> when did they introduce this? I must have missed it...

> https://www.visitron.de/products/visiscope-confocal/yokogawa-sora.html

> I know Yokogawa engineers published it back in 2015, but I did not

> know it's finally available.

>

> Has anyone tried/purchased it? What are the impressions?



We played with one on an Olympus setup. It is slightly different than the standard Yokogawa setup as they have their own beam expansion optics and then the off the shelf SORA disk.



Seems to work pretty well. Definitely getting super-resolution data but not convinced by the Olympus weiner filtering and deconvolution idea. Also seems very expensive in the UK, ball park price seems to be

£350-500,000 depending on options.



It is easy to go from conventional spinning disk to the SORA mode, and the lenses in front of the pinholes mean you get significantly more light through. The PSF on thick samples can be pretty horrific though. I think it would be very hard to deconvolve in bright thick samples as the out of focus bleed through from other excitation spots is very image position dependant, you can see that arc's due to the pinhole positions on the disk.



Ian



När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://ki.se/medarbetare/integritetsskyddspolicy>.


Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI’s processing of personal data here<https://ki.se/en/staff/data-protection-policy>.

[Ian Dobbie]

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Sylvie Le Guyader <[hidden email]> writes:

> I have a couple of questions:
>
> - Is the price you mention for an upgrade (i.e. the Olympus microscope
> was yours) or is it for a full system with lasers, camera…?

The price was for a complete system, obviously exact price depends on
configuration and discounts etc....

> - When you talk about? Do you think that the pinhole crosstalk you
> mention is worse with SoRa than with the conventional/standard
> Yokogawa spinning disk (by ‘conventional/standard’, do you mean W1 or
> X1?)? We have an X1 we are very happy with but if we don’t clear the
> sample and we image thick samples, we get the expected pinhole
> crosstalk.

The crosstalk I mention is on the SoRa disk with the 3x magnifier in
front to get the super resolution images. The normal W1 fairs much
better than the older X1 or CSU22 as the pinholes are further apart.

My feeling for the crosstalk, although not measured, is that the
crosstalk on the SoRa disk happens at much lower depths, but is less
intense relative to the main through pinhole image compared to the older
X1's that we have. I'm afraid I haven't put the same sample on both
instruments so this is just a feeling.

Ian

[lgelman]

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Hi,

I agree with Emmanuel.
On our W1 systems (see my last mail :-) we have also Homogenizers from Visitron.
The fact is that you cannot get full homogeneity for all combinations of magnifications and wavelengths. It can be perfect for a given combination if you need it absolutely, but then very suboptimal for others, so usually the way to go is to find a compromise. We also see variations form 5 to 30%.
Best regards,

Laurent.


Laurent Gelman
Facility for Advanced Imaging and Microscopy
Head Light Microscopy
Friedrich Miescher Institute for Biomedical Research
Maulbeerstrasse 66
4058 basel
Switzerland
[phone]




-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Emmanuel Levy
Sent: Thursday, February 21, 2019 10:43
To: [hidden email]
Subject: Re: Yokogawa W1 spinning disk with OPRA-style superresolution

*****
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,

I have no experience with the SORA add-on. One thing to keep in mind is that it decreases the field of view by 75% (because of the 2x magnification).

On a different note, anyone familiar with the Visitron's beam homogenizing
> system for CSU-W1 willing to share some details? How does it compare
> to "Andor's" Borealis? The thing is that we're observing some strange
> borealis-related effects at short exposures; moreover, Andor does not
> sell
> W1 with borealis anymore...
>

We've had the homogenizer for a while. The homogeneity is better than with regular W1 optics but not flat. In our system we get ~10-15% variation across the FOV for GFP, 20-25% for RFP, and about or above 30% for BFP.

Hope this helps,

Emmanuel

>
> Thanks!
>
> Best, zdenek
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Associate - Imaging Specialist Department of Biochemistry and
> Cell Biology Geisel School of Medicine at Dartmouth
>