Posted by
John Oreopoulos on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Johannes-Amon-posted-new-Zeiss-white-paper-on-airyscanning-at-tp7582420p7582422.html
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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 <
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 <
https://www.linkedin.com/profile/view?id=86600201&goback=%2Egde_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
>
http://www.optnano.com/content/2/1/7/abstract>
> For an entry point to SOFI see
>
>
http://www.optnano.com/content/2/1/2/abstract>
> As I've noted on the Confocal Listserv
>
>
http://lists.umn.edu/cgi-bin/wa?A2=ind1407&L=CONFOCALMICROSCOPY&P=5920>
> I am psyched what parallel processing is doing (ex. NVidia TITAN Z GPU 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