Optical slice thickness and number for PSF and deconvolution

[Martin Wessendorf-2]

Hey, Guy--

Not to drag this out any longer, but could you please clarify?  I
understand that intensity comparisons from deconvolved image to
deconvolved image might be problematic.  However, the statement that
deconvolved images are not quantifiable strikes me as incorrect.  As I
see it, the goal of deconvolution is to provide a more accurate idea of
the relative concentration of fluorophore within different compartments
of a cell--the image is less contaminated by out-of-focus information.

Thanks.

Hope all is well in Sydney--

Martin

On 8/17/2010 3:46 AM, Guy Cox wrote:

> Yes, your post came through.  I didn't comment because at the time I was
> embroiled in an off-list debate with Mark Cannell on the same topic.
> (It seems our differences were largely terminology - he regards setting
> the pinhole at 1 Airy unit to be opening it up whereas I regard that as
> keeping it closed!)
>
> As to your post, I just cannot accept that deconvolution is 'essential'.
> It may give you a prettier picture, and at best it may well show you
> things that you couldn't see before, which has obvious value.  But the
> downside is that you end up with an image that is non-quantifiable, and
> to me that is a big sacrifice.  I use deconvolution, and have even
> written simple deconvolution software, but I cannot accept that it is
> essential.
>
>                                             Guy

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