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September 2009

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From:
Jose Viña <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Tue, 22 Sep 2009 14:42:13 +0200
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Hi Sundar,

> I'm using a deconvolution software to deconvolve a 3D stack. Does
> anyone know of any way in which I can measure the improvement over
> the original dataset? Are there any standard techniques for the same?

There are various kinds of 'improvement' deconvolution can bring you.
If you are interested in improving half intensity width (HIW) of
particles, best follow Louis' suggestion to measure HIW in lateral and
axial of beads. You will almost certainly learn interesting things
about your microscope.

If on the other hand you are working with widefield images and blur is
hampering your analysis, you might be only interested in reducing the
blur. Question is then how much reduction you need. A test object
could be a larger bead, but beads can have a high internal refractive
index, upsetting things.

Lastly, you might be working with noisy but well sampled data and
trying to measure object shapes. Then I would suggest to construct
synthetic objects and simulate the whole imaging, Poisson noise,
deconvolution and measurement chain, and see what the effect is on the
error in the measurement you plan to do.

Some links you might find useful:

A quantitative comparison of two restoration methods as applied to
confocal microscopy.
van Kempen GMP, van der Voort HTM and van Vliet LJ
http://www.svi.nl/g/?7028

A comparison of image restoration approaches applied to
three-dimensional confocal and wide-field fluorescence microscopy.
P. J. Verveer, M. J.  Gemkow and T. M.  Jovin.
http://www.svi.nl/g/?71a3

I hope this helps!

Cheers,

jose.

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