Posted by
Brian Northan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/confocal-detectors-and-deconvolution-tp7588223p7588243.html
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>
> and yes there has to be some care taken so that artifacts (present in raw
> data, ie SA)
Hi Rich
Artifacts in deconvolved images, caused by aberrated PSF, can be avoided
to some extent, by using an accurate PSF. Either a high quality measured
PSF, or (atleast for SA) a theoretical PSF (ie Gibson Lanni) with good
estimates for Refractive Index of the Specimen, and Imaging depth. Mike
wrote a paper, a few years ago, on PSF generation and deconvolving
aberrated images (
https://www.ncbi.nlm.nih.gov/pubmed/21118213).
Brian
On Fri, May 11, 2018 at 9:17 AM, Cole, Richard W (HEALTH) <
[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.
> *****
>
> Mike,
>
> I have two HyDs and I agree w/Vitaly, there is essentially no difference
> when compared to GaAsP. We run almost all imaging modalities through
> deconvolution (Autoquant GPU), and yes there has to be some care taken so
> that artifacts (present in raw data, ie SA) are not accentuated. As for
> quantitative decon see: Calibration of wide-field deco
> nvolution microscopy for quantitative fluorescence imaging. Lee JS; Wee
> TL; Brown CM. J. of Biomol. Tech. 25(1):31-40, 2014 Apr.
>
> My 2¢
>
>
> Richard Cole
> Research Scientist V
> Director: Advanced Light Microscopy & Image Analysis Core
> Wadsworth Center
>
> Research Assistant Professor
> Dept. of Biomedical Sciences
> School of Public Health State University of New York
>
> 120 New Scotland Avenue, Albany N.Y. 12208
> 518-474-7048 Phone
> 518-408-1730 Fax
>
>
>
>
>
> >Hi Mike,
> >there is no big difference between HyDs and GaAsP detectors. Leica SMD
> HyDs are cooled to 18 C, thus having ca. 400 dark counts versus 2K or more
> for the GaAsP.I am >not aware of linear deconvolution algorithms for
> confocal image data, thus it seems to be "cosmetic" and/or "aesthetic", and
> not suitable for accurate intensity based >image analysis.Recent
> integration of FALCON combined with FCS/RICS to the Leica SP8 may bring new
> era to biology letting biologists quantify their data and image >biological
> molecules at their physiological concentrations. However, most of
> biologists seems to be "scared" of basic equations...If you are located in
> the US, Leica provides >much better Service and Application support
> compared to Zeiss. I do not have much experience with Olympus and Nikon on
> the confocal side.
> >If you have any specific questions, please let me know.
> >VitalyMCCF/MSKCC
>
>
http://www.lsoft.com/products/listserv-powered.asp>
On Fri, May 11, 2018 at 9:17 AM, Cole, Richard W (HEALTH) <
[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.
> *****
>
> Mike,
>
> I have two HyDs and I agree w/Vitaly, there is essentially no difference
> when compared to GaAsP. We run almost all imaging modalities through
> deconvolution (Autoquant GPU), and yes there has to be some care taken so
> that artifacts (present in raw data, ie SA) are not accentuated. As for
> quantitative decon see: Calibration of wide-field
> deconvolution microscopy for quantitative fluorescence imaging. Lee JS;
> Wee TL; Brown CM. J. of Biomol. Tech. 25(1):31-40, 2014 Apr.
>
> My 2¢
>
>
> Richard Cole
> Research Scientist V
> Director: Advanced Light Microscopy & Image Analysis Core
> Wadsworth Center
>
> Research Assistant Professor
> Dept. of Biomedical Sciences
> School of Public Health State University of New York
>
> 120 New Scotland Avenue, Albany N.Y. 12208
> 518-474-7048 Phone
> 518-408-1730 Fax
>
>
>
>
>
> >Hi Mike,
> >there is no big difference between HyDs and GaAsP detectors. Leica SMD
> HyDs are cooled to 18 C, thus having ca. 400 dark counts versus 2K or more
> for the GaAsP.I am >not aware of linear deconvolution algorithms for
> confocal image data, thus it seems to be "cosmetic" and/or "aesthetic", and
> not suitable for accurate intensity based >image analysis.Recent
> integration of FALCON combined with FCS/RICS to the Leica SP8 may bring new
> era to biology letting biologists quantify their data and image >biological
> molecules at their physiological concentrations. However, most of
> biologists seems to be "scared" of basic equations...If you are located in
> the US, Leica provides >much better Service and Application support
> compared to Zeiss. I do not have much experience with Olympus and Nikon on
> the confocal side.
> >If you have any specific questions, please let me know.
> >VitalyMCCF/MSKCC
>
>
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