PSF

[Bastien Pachoud]

Dear all,

I am trying to use the Diffraction PSF 3D plugin and I have several
questions.

1- How can I find or measure the longitudinal spherical aberration value
of my optical system?

2- Does the Image pixel spacing correspond to the size of the pixel?

3- I want to use deconvolution to deblurr 2D (one single frame)
fluorescence images. How should I define the number of slices needed for
my PSF?

thank you very much in advance...

regards,

Bastien

[Robert Dougherty]

Bastien,

On Aug 6, 2010, at 3:38 AM, Bastien Pachoud wrote:

> Dear all,
>
> I am trying to use the Diffraction PSF 3D plugin and I have several questions.
>
> 1- How can I find or measure the longitudinal spherical aberration value of my optical system?

If you have to ask, it means you can't measure it directly.  This would normally be a job for specialized laboratory.  It is probably best to enter 0. In principle,  you could try different values to see if some value works better than another for your microscope in terms of matching the image from a bead or giving good deconvolution results. I do not recommend this with your 2D images.

>
> 2- Does the Image pixel spacing correspond to the size of the pixel?

As noted in the dialog, it is the ccd cell spacing divided by the magnification.
>
> 3- I want to use deconvolution to deblurr 2D (one single frame) fluorescence images. How should I define the number of slices needed for my PSF?

Choose 1 slice if you want to do 2D deconvolution.
>
> thank you very much in advance...
>
> regards,
> Bastien

Robert Dougherty, Ph.D.
President, OptiNav, Inc.
4176 148th Ave. NE
Redmond, WA 98052
Tel. [phone]
FAX [phone]
www.optinav.com
[hidden email]

[Cuculus]

Hello! I am a beginner with ImageJ and I need to do 3D deconvolution for a stack of images, but I don't understand what ccd cell spacing is. I will appreciate some help.

Thanks in advance!

[Elena Kutsarova]

Dear all,

Does somebody know what the difference between the two plugins: 3D Diffraction PSF (Robert Dougherty) and PSF Generator (http://bigwww.epfl.ch/deconvolution/?p=plugins) is?

Thanks very much!