Posted by
Zdenek Svindrych on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Johannes-Amon-posted-new-Zeiss-white-paper-on-airyscanning-at-tp7582420p7582423.html
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Very interesting.
It's most likely based on the Image Scanning Microscopy (ref 4 of the white
paper; it's a physics journal, so the style is a bit different), it allows
up to 2x resolution improvement (as all the other linear structured
illumination methods), but with the Enderlain's image processing you
actually sacrifice axial resolution if you want high SNR (corresponds to
bigger pinhole). Unfortunately they don't provide much details in the white
paper about the new image processing approach that gives also axial
superresolution...
Moreover, I think this approach is not quite compatible with other benefits
of modern confocals, such as spectral detection or lifetime imaging.
Looking forward to seeing further advances in this direction.
Zdenek Svindrych, Charles univ., Czech Rep.
---------- Původní zpráva ----------
Od: John Oreopoulos <
[hidden email]>
Komu:
[hidden email]
Datum: 25. 7. 2014 4:30:32
Předmět: Re: Johannes Amon posted new Zeiss white paper on airyscanning at .
..
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The white paper states:
"An acentric, shifted pinhole detector produces an image of about the same
resolution as a pinhole detector which is aligned to the optical axis,
although smaller in amplitude and shifted by half the displacement. This
insight has been the motivation for constructing an area detector for a
confocal microscope. Such a detector should cover more than 1 AU and contain
multiple sub-Airy detector elements. Detection efficiency will be
significantly increased by reassigning the detected photons from the shifted
detector elements to the central detection position and summing up the back
shifted signal from all detector elements. [2] No light is rejected by a
closed pinhole but instead collected by the off-axis detector elements.
Therefore an increased signal level arises from the reassignment of photons
to a smaller spatial region."
It sure does sound a lot like Jim Pawley's array detector pinhole idea to
me, which is outlined in quite clearly chapter 2 of the Handbook of
Biological Confocal Microscopy (3d Edition):
http://link.springer.com/chapter/10.1007/978-0-387-45524-2_2See Figure 2.15 in particular. There isn't much detail in the white paper to
know what the differences are however, and there is no citation of Pawley's
work on this. Here's the relevent passage from the chapter:
"In addition to high QE, Si photon detectors have a variety of other
practical advantages. As the sensitive element in such a detector is
typically very small (5–30 mm on a side), selective use of only a few
elements in a small, planar, 2D array could permit it to operate in the CLSM
as a combination pinhole and detector. Figure 2.15 is a sketch of what such
a detector might look like. After each pixel interval of the microscope, the
charge pattern in the 5 x 5 sensor array at the top would be transferred to
the read register and then the signal in all 25 pixels would be read out
sequentially at about 35MHz. These 25 values could then be “decoded” in a
number of possible ways, the most straightforward of which would be to
provide three separate signals corresponding to the summed signals from the
brown, orange, and red areas of the sensor array. In this way, it would be
possible to collect signal simultaneously at three different pinhole sizes.
With such a detector, pinhole alignment could be done elec- tronically
simply by searching for the detector element producing the most signal from
a planar specimen and misalignment could be detected on the fly by
comparing, for example, summed output from the 5 pixels on the left with the
5 on the right (Pawley, 1996)."
The other reference from 1996 has even more details about this idea:
CCDiode: an optimal detector for laser confocal microscopes
http://spie.org/Publications/Proceedings/Paper/10.1117/12.237490John Oreopoulos
On 2014-07-24, at 8:57 PM, George McNamara 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.
> *****
>
> Johannes Amon posted the URL to download a new Zeiss white paper on
airyscanning
>
>
http://www.zeiss.com/airyscan <<a href="https://www.linkedin.com/redirect?url=http%">https://www.linkedin.com/redirect?url=http%
3A%2F%2Fwww%2Ezeiss%2Ecom%2Fairyscan&urlhash=fM6m&_t=tracking_anet>
>
> Johannes post is at
>
>
https://www.linkedin.com/groupAnswers?viewQuestionAndAnswers=&discussionID=5898093815864004612&gid=837267
>
> and he welcomes comments.
>
>
> Airyscanning: A Novel Approach to Confocal Imaging
>
> Johannes Amon <<a href="https://www.linkedin.com/profile/view?id=86600201&goback=%2">https://www.linkedin.com/profile/view?id=86600201&goback=%2
Egde_837267_member_5898093815864004612>Online Communications and Marketing
>
> It is with great pleasure that I am able to share with you a new White
Paper by ZEISS. Airyscanning is a new detection concept that uses an array
detector to oversample each Airy disk in order to gain sensitivity,
resolution and speed. I'm curious to hear what you think of it! Download the
free White Paper here:
http://www.zeiss.com/airyscan <
https://www.linkedin.
com/redirect?url=http%3A%2F%2Fwww%2Ezeiss%2Ecom%2Fairyscan&urlhash=fM6m&_t=
tracking_anet>
>
>
> Vladimir Zhukarev and I have already commented at the linkedin group. My
comments are:
>
>
>
> Zeiss cites some but not all of the literature of using a (small) area
array to collect light that would usually be blocked by the pinhole. I'm
thinking that Jim Pawley had this, but could be 'off by one' (Tony Wilson,
Guy Cox, etc?).
>
> Too bad Zeiss and the other microscope companies have pathetically slow
spatial deconvolution AND on confocal microscopes deconvolution is a
separate post-processing step the user has to undertake manually (I managed
Leica confocals since 2000 and Zeiss LSM710 since 2009 ... just trained this
week on a Leica SP8 confocal, LAS AF 3.x, haven't seen all the features, but
deconvolution looks like still a separate manual step).
>
> Further, to get the maximum benefit from a confocal microscope, when
acquiring multiple channels, should BOTH spatially deconvolve AND spectrally
unmix. See the Hoppe et al 2008 Biphys J paper,
>
>
http://www.ncbi.nlm.nih.gov/pubmed/18339754>
> their 3DFSR was tested with FRET specimens but the (claimed) 10x improved
SNR should apply to any specimens.
>
> I also note that the Zeiss (claimed) 1.7x improvement could be massively
improved by acquiring 300 images (per plane) and doing 3B microscopy or SOFI
- for 3B see
>
>
http://www.coxphysics.com/3b/>
> for introduction to 3B (and yes, could work in 3D). 3B is also very slow.
Parallel processing is speeding up 3B, see
card, Intel Phi 7120 card, lots of Phi's, as at
https://www.tacc.utexas.edu/stampede/ ) and will get even better with 2015 Knights Landing CPU's (and
NVidia, IBM, HP if its "The Machine" meets their goals, etc).
>
> George
>
>
> www.linkedin.com/in/georgemcnamara/
>
> --
>
>
>
>
> George McNamara, Ph.D.
> Single Cells Analyst
> L.J.N. Cooper Lab
> University of Texas M.D. Anderson Cancer Center
> Houston, TX 77054
> Tattletales
http://works.bepress.com/gmcnamara/42"