Lightsheet z.1 vs ultramicroscope

[Vladimir Ghukasyan-2]

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Dear Listers,

We are considering getting a light sheet setup for our core. I had a chance
to test briefly the Z.1 system from Zeiss and prepare to send some samples
to Germany to test the Ultramicroscope. Havy any of you worked with either
of these systems? I would be very thankful for any suggestion/comment
advice on this matter.

Our typical application is imaging of CLARITY cleared mouse brain samples.
High resolution may be very beneficial (imaging of axonal tracks) but not
absolutely necessary at this point. The Z.1 can work with a "CLARITY" long
working distance objectives, but the immersion media is limited to
non-organic solvents, so no BAAB/iDISCO, etc. Ultramicroscope as I have
learned today, can be used with any objective, but I wonder how is
chromatic aberration correction handled (hope to hear about that from the
company soon).

Thank you in advance,
Vladimir

[Bernd Mueller-Zuelow]

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For all of those who are not really close to this field a little excerpt from the recent literature.



Whole-Brain
Imaging with Single-Cell Resolution Using Chemical Cocktails and Computational
Analysis. http://www.ncbi.nlm.nih.gov/pubmed/24746791 (Cell) Comments: Cubic, GFP

iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume
Imaging. http://dx.doi.org/10.1016/j.cell.2014.10.010 (Cell) Comments: many organs,
iDISCO

A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent
Nervous System. http://dx.doi.org/10.1016/j.celrep.2014.10.037 (Cell) Comments:
iDISCO, axonal guidance

Optimization of CLARITY for Clearing Whole-Brain and Other Intact Organs. http://eneuro.org/content/2/3/ENEURO.0022-15.2015
(ENEURO) Comments: Clarity


Many more publications on different clearing methods and the use of the Ultramicroscope system can be found at http://www.lavisionbiotec.com/ultramicroscope-literature.html



Best regards

Bernd Mueller-Zuelow

--

LaVision BioTec GmbH

Dr. Bernd Mueller-Zuelow

Area Sales Manager

[hidden email]

> Date: Mon, 29 Jun 2015 17:55:41 -0400
> From: [hidden email]
> Subject: Lightsheet z.1 vs ultramicroscope
> To: [hidden email]
>
> *****
> 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,
>
> We are considering getting a light sheet setup for our core. I had a chance
> to test briefly the Z.1 system from Zeiss and prepare to send some samples
> to Germany to test the Ultramicroscope. Havy any of you worked with either
> of these systems? I would be very thankful for any suggestion/comment
> advice on this matter.
>
> Our typical application is imaging of CLARITY cleared mouse brain samples.
> High resolution may be very beneficial (imaging of axonal tracks) but not
> absolutely necessary at this point. The Z.1 can work with a "CLARITY" long
> working distance objectives, but the immersion media is limited to
> non-organic solvents, so no BAAB/iDISCO, etc. Ultramicroscope as I have
> learned today, can be used with any objective, but I wonder how is
> chromatic aberration correction handled (hope to hear about that from the
> company soon).
>
> Thank you in advance,
> Vladimir

     

[Alison J. North]

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Hi Vladimir,

We have a LaVision Ultramicroscope and I would be happy to answer any
specific questions you may have about it if you want to contact me
offline.  I'm not sure from your e-mail what kind of suggestions or
comments you are looking for.
It might also be helpful if you could say where you are located, in case
you have questions about service or whatever - that's always easier to
tell from a work e-mail address than from a gmail one...

All the best,
Alison

On 6/29/2015 5:55 PM, Vladimir Ghukasyan 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 Listers,
>
> We are considering getting a light sheet setup for our core. I had a chance
> to test briefly the Z.1 system from Zeiss and prepare to send some samples
> to Germany to test the Ultramicroscope. Havy any of you worked with either
> of these systems? I would be very thankful for any suggestion/comment
> advice on this matter.
>
> Our typical application is imaging of CLARITY cleared mouse brain samples.
> High resolution may be very beneficial (imaging of axonal tracks) but not
> absolutely necessary at this point. The Z.1 can work with a "CLARITY" long
> working distance objectives, but the immersion media is limited to
> non-organic solvents, so no BAAB/iDISCO, etc. Ultramicroscope as I have
> learned today, can be used with any objective, but I wonder how is
> chromatic aberration correction handled (hope to hear about that from the
> company soon).
>
> Thank you in advance,
> Vladimir

--
Alison J. North, Ph.D.,
Senior Director of the Bio-Imaging Resource Center and
Research Associate Professor,
The Rockefeller University,
1230 York Avenue,
New York,
NY 10065.
Tel: office ++ 212 327 7488
Tel: lab    ++ 212 327 7486
Fax:        ++ 212 327 7489

[Michael Weber-4]

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Hi Vladimir,

LightSheet Z.1 and Ultramicroscope are two very different microscopes. The first could be a replacement for a spinning disc confocal, whereas the latter is closer to a point-scanning confocal. It really depends on what kind of specimen you are using.

The Zeiss system is designed for fast imaging of living samples, multi-view recordings, time-lapse imaging. Optics are optimized for the refractive index of water, the specimen can be heated and supplied with CO2. The sample is hanging from top and can be rotated by 360 deg, therefore it must be mounted in a suitable manner.

The LaVision system is made for fixed, cleared samples. It is not overly fast and the specimen cannot be rotated. However, because of the different design you can use all kinds of solvents, which might be essential for recordings of thick tissues or large specimen.

Best,
Michael


On Jun 29, 2015, at 11:55 PM, Vladimir Ghukasyan <[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 Listers,
>
> We are considering getting a light sheet setup for our core. I had a chance
> to test briefly the Z.1 system from Zeiss and prepare to send some samples
> to Germany to test the Ultramicroscope. Havy any of you worked with either
> of these systems? I would be very thankful for any suggestion/comment
> advice on this matter.
>
> Our typical application is imaging of CLARITY cleared mouse brain samples.
> High resolution may be very beneficial (imaging of axonal tracks) but not
> absolutely necessary at this point. The Z.1 can work with a "CLARITY" long
> working distance objectives, but the immersion media is limited to
> non-organic solvents, so no BAAB/iDISCO, etc. Ultramicroscope as I have
> learned today, can be used with any objective, but I wonder how is
> chromatic aberration correction handled (hope to hear about that from the
> company soon).
>
> Thank you in advance,
> Vladimir

_____________

Michael Weber
Postdoc, Huisken lab
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108, 01307 Dresden
Tel. [phone]

http://www.mpi-cbg.de/huisken

[Douglas Richardson]

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Hi All,

I think I would disagree with the comparisons to a spinning disk and point
scanner.  The Z1 is not a replacement for a spinning disk, as it will not
achieve the same Z resolution (1.0NA optics versus 1.4) and the lightsheet
cannot be focused as thin as needed to compensate for the difference in
NA.  However, at lower magnifications, the Z1 will out perform a
traditional spinning disk in which the disk was designed specifically for
high NA objectives and often paired with a large pixel size EMCCD camera.
The Z1 will also outperform the spinning disk in imaging depth, especially
when utilizing multi-view.

I certainly would not compare the Ultramicroscope to a point scanning
confocal as it is a low resolution system - the exact opposite of a point
scanner.  It uses low NA optics (especially at low zoom/large field of view
settings) and although it has a 4MP sCMOS, it is still undersampling at
these zoom levels. Also, to cover this large field of view, you cannot
focus your lightsheet very thin, therefore you cannot use this to
compensate for the poor Z-resolution of your low NA imaging objective

My take:

Advantages of Ultramicroscope:

Large field of view: fast, small file size
Easy sample mounting
Compatible with solvent clearing techniques
Price

Disadvantages:

Low resolution: Can be improved with higher zoom levels and tiling, but
still not as good as high NA dipping optics on confocals or Z1

Advantages of Z1:

Capable of live imaging, multiview
Higher resolution (1.0NA dipping optics for aqueous samples, 1.38 or 1.45
RI clearing solutions)

Disadvantages:

Large file sizes
Slower for large cleared samples (always requires tiling)
Price
Not compatible with solvent clearing (although I know a few groups have
developed new sample chambers or inserts that protect the plastic
components of the microscope; these are still not compatible with the high
NA dipping objectives)

-Doug


On Thu, Jul 2, 2015 at 3:59 AM, Michael Weber <[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.
> *****
>
> Hi Vladimir,
>
> LightSheet Z.1 and Ultramicroscope are two very different microscopes. The
> first could be a replacement for a spinning disc confocal, whereas the
> latter is closer to a point-scanning confocal. It really depends on what
> kind of specimen you are using.
>
> The Zeiss system is designed for fast imaging of living samples,
> multi-view recordings, time-lapse imaging. Optics are optimized for the
> refractive index of water, the specimen can be heated and supplied with
> CO2. The sample is hanging from top and can be rotated by 360 deg,
> therefore it must be mounted in a suitable manner.
>
> The LaVision system is made for fixed, cleared samples. It is not overly
> fast and the specimen cannot be rotated. However, because of the different
> design you can use all kinds of solvents, which might be essential for
> recordings of thick tissues or large specimen.
>
> Best,
> Michael
>
>
> On Jun 29, 2015, at 11:55 PM, Vladimir Ghukasyan <[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 Listers,
> >
> > We are considering getting a light sheet setup for our core. I had a
> chance
> > to test briefly the Z.1 system from Zeiss and prepare to send some
> samples
> > to Germany to test the Ultramicroscope. Havy any of you worked with
> either
> > of these systems? I would be very thankful for any suggestion/comment
> > advice on this matter.
> >
> > Our typical application is imaging of CLARITY cleared mouse brain
> samples.
> > High resolution may be very beneficial (imaging of axonal tracks) but not
> > absolutely necessary at this point. The Z.1 can work with a "CLARITY"
> long
> > working distance objectives, but the immersion media is limited to
> > non-organic solvents, so no BAAB/iDISCO, etc. Ultramicroscope as I have
> > learned today, can be used with any objective, but I wonder how is
> > chromatic aberration correction handled (hope to hear about that from the
> > company soon).
> >
> > Thank you in advance,
> > Vladimir
>
> _____________
>
> Michael Weber
> Postdoc, Huisken lab
> Max Planck Institute of Molecular Cell Biology and Genetics
> Pfotenhauerstrasse 108, 01307 Dresden
> Tel. [phone]
>
> http://www.mpi-cbg.de/huisken
>