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June 2011

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From:
Mark Bates <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Mon, 27 Jun 2011 05:48:07 -0700
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hi all,

Thanks Jim for your synopsis of the related work in the EM field - I wasn't
aware of that.  Just to touch on a couple of the points raised, I agree with
Jim regarding the statistics of switchable molecules - this can result in
the STORM image becoming non-linear with respect to how the intensity in the
image corresponds to the local concentration of fluorophores on the sample. 
This all depends on the nature of the switchable fluorophore of course,
whether it activates once and then bleaches, or if it's reversibly
switchable, etc.  In principle it would be very useful to treat these images
as quantitative maps of fluorophore position and concentration, and this is
where one needs to be careful in considering the switching statistics of the
fluorophore.

Regarding the name debate, PALM, STORM, FPALM etc all refer to precisely the
same concept.  PALM should not be associated strictly with proteins, nor
should STORM be associated with one type of fluorescent probe.  We made it
clear in our original paper that the concept is applicable to any switchable
fluorophore.  I don't see the need for the acronyms which have been
subsequently coined, except where there is new science involved (such as
GSDIM, which extends the concept to any photophysical dark state).

best,
Mark Bates, Ph.D.
Max Planck Institute for Biophysical Chemistry
Göttingen 37077
Germany 

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