Re: Lightsheet z.1 vs ultramicroscope

Posted by Douglas Richardson on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Lightsheet-z-1-vs-ultramicroscope-tp7583940p7583970.html

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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:

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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:
>
> > *****
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> > 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
>