Re: High speed spinning disc confocal with EMCCD camera - commercial response

Posted by George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/High-speed-spinning-disc-confocal-with-EMCCD-camera-tp7583142p7583219.html

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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; /
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<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