Posted by
Andreas Bruckbauer on
URL: http://confocal-microscopy-list.275.s1.nabble.com/High-speed-spinning-disc-confocal-with-EMCCD-camera-tp7583142p7583233.html
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Hi,
This has gone slightly off topic now, but Nyquist might be the next barrier to fall, see compressed sensing which has already been used in microscopy and I think looks very promising:
Faster STORM using compressed sensing.
Zhu L, Zhang W, Elnatan D, Huang B.
Nat Methods. 2012 Apr 22;9(7):721-3. doi: 10.1038/nmeth.1978.
Single-shot compressed ultrafast photography at one hundred billion frames per second
Liang Gao,
Jinyang Liang,
Chiye Li
& Lihong V. Wang
Nature 516, 74–77 (04 December 2014)
Andreas
-----Original Message-----
From: "Michael Giacomelli" <
[hidden email]>
Sent: 06/01/2015 19:47
To: "
[hidden email]" <
[hidden email]>
Subject: Re: High speed spinning disc confocal with EMCCD camera - commercial response
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Hi George,
You probably mean "diffraction" rather than "Nyquist". The Nyquist limit
applies even to superresolution systems, whereas the diffraction limit does
not. Guy's point is that one only needs to sample densely enough to
satisfy the Nyquist critiera (for a diffraction or subdiffraction) system.
Going further helps SNR (it is really just a form of averaging) but does
nothing for resolution.
Mike
On Tue, Jan 6, 2015 at 12:28 AM, George McNamara <
[hidden email]>
wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
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>
> Hi Guy,
>
> Nyquist is dead.
>
> For mitochondria way beyond, see Shim et al 2012,
>
http://www.ncbi.nlm.nih.gov/pubmed/22891300>
http://www.pnas.org/content/109/35/13978.long (open access)
>
> also several papers from Hell and colleagues, such as;
>
>
http://www.ncbi.nlm.nih.gov/pubmed/20205711>
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2838807/pdf/1757-5036-3-4.pdf>
> and
> 30 nm isotropic resolution (isoSTED)
>
http://www.ncbi.nlm.nih.gov/pubmed/19459703>
> George
> p.s.
http://en.wikipedia.org/wiki/Harry_Nyquist> *Harry Nyquist* (/né/ Harry Theodor Nyqvist; / <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English>ˈ <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>n <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>aɪ <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>k <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>w <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>ɪ <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>s <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>t <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English#Key>/ <
>
http://en.wikipedia.org/wiki/Help:IPA_for_English>, Swedish: [nʏːkvɪst] <
>
http://en.wikipedia.org/wiki/Help:IPA_for_Swedish_and_Norwegian>;
> February 7, 1889 – April 4, 1976)
>
> On 1/5/2015 10:10 AM, Guy Cox wrote:
>
>> Michael,
>>
>> Both you and I have been telling users this for years.
>> Likewise, if they want to image a mitochondrion, they gain nothing by going
>> beyond Nyquist, yet they still zoom it up to 1024 x 768. These people are
>> supposed to be scientists - but are they? A microscope is a scientific
>> instrument, not a black box.
>>
>> Guy
>>
>> Guy Cox, Honorary Associate Professor
>> School of Medical Sciences
>>
>> Australian Centre for Microscopy and Microanalysis,
>> Madsen, F09, University of Sydney, NSW 2006
>>
>>
>> -----Original Message-----
>> From: Cammer, Michael [mailto:
[hidden email]]
>> Sent: Tuesday, 6 January 2015 2:57 AM
>> To: Guy Cox
>> Subject: RE: High speed spinning disc confocal with EMCCD camera -
>> commercial response
>>
>> I constantly tell people doing live imaging with the multiphoton who want
>> to track cells to turn down the excitation and do running averages or other
>> noise filtering later. I show them examples but when I leave the room they
>> invariably crank up the light because they want to see a perfect low noise
>> high contrast image right now and then they are disappointed when the
>> experiment doesn't work either because the sample bleaches or the cells
>> stop moving.
>> =========================================================================
>> Michael Cammer, Microscopy Core& Skirball Institute, NYU Langone
>> Medical Center
>> Cell: 914-309-3270 Temporary location:
>> SK2-7
>>
http://ocs.med.nyu.edu/microscopy&>>
http://microscopynotes.com/>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List [mailto:
[hidden email]]
>> On Behalf Of Guy Cox
>> Sent: Monday, January 05, 2015 10:45 AM
>> To:
[hidden email]
>> Subject: Re: High speed spinning disc confocal with EMCCD camera -
>> commercial response
>>
>>
>> Not being able to saturate excitation is a good thing, not a bad thing!
>> I am perpetually telling people to reduce their excitation to 50% and check
>> that their excitation decreases by the same amount - if not they are
>> saturating (and destroying) their fluorochrome. Nobody ever follows my
>> advice - they just want a bright (and usually saturated and therefore
>> uninformative) image.
>>
>> Guy
>>
>>
>> Guy Cox, Honorary Associate Professor
>> School of Medical Sciences
>>
>> Australian Centre for Microscopy and Microanalysis, Madsen, F09,
>> University of Sydney, NSW 2006
>>
>>
>>
>>
>
>
> --
>
>
>
> 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>