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February 2017

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From:
JAMES B PAWLEY <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Tue, 14 Feb 2017 01:57:34 +0000
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Hi all,

Those attending the sadly-gone UBC 3D-course may remember that our first  slogan was, "If it isn’t diffraction, it’s statistics.”

Colin’s wonderful, inclusive paper does indeed wrap up the diffraction part superbly.

But how many photons/nm*3 would you have to collect in order to obtain data that would allow you to fill in those beautifully narrow lines? Millions? (if the line width is say 1% of its “value” surely you need 10,000 detected photons to determine it  And how big must a pixel be to trace out these lovely curves without them looking blocky? Given that biologists habitually work with specimens capable of producing at most tens of detectable photons/pixel, surely it is the statistics that places the most severe spatial limits on the data we finally obtain.

Where is the young mathematician (statistician?) who can give us an equally comprehensive analysis of the number of photons  per cubic nm that we must elicit and detectin order to “resolve” two closely spaced point objects? And why do we always assume that these two objects have the same intensity when this is so seldom the case? What visibility criteria would be appropriate to model the problem of seeing the Earth from Alpha Centauri?

Please someone take up this quest.

Jim Pawley

James and Christine Pawley, 5446 Burley Place, Box 2348, Sechelt BC, Canada, V0N3A0 [log in to unmask]<mailto:[log in to unmask]>, Phone 1-604-885-0840, cell 1-604-989-6146



On Feb 13, 2017, at 7:11 AM, Alberto Diaspro <[log in to unmask]<mailto:[log in to unmask]>> wrote:

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Dear
following a discussion with the MRT staff and Wiley, they informed me that the paper is now available for free for one month. Hope this can help.
Please, have a check and let me know if it works at http://onlinelibrary.wiley.com/doi/10.1002/jemt.22834/full

Best
Alby


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