Point spread function measurement and hyperstack

[Pang, Zhengyu (GE Global Research)]

Dear all,

I obtained a series of z plane images of beads, a tiff file with 35
slices, and need to compute the point spread function (PSF) for Z plane.
I could crop original image to make only one bead in the field.

I plan to threshold each cropped image (or make it binary) to identify
the signal region, and then calculate the average singal intensity. Next
I want to calculate the background intensity by dilating the binary
images and invert the selection to calculate background intensity of
grayscale images.

Intensity for PSF will be calculated as (MeanSignal -Mean
Background)/3/stdev of Background.

I think I need to use two sets of images (raw grayscale image and
binary mask image to make selection) But I am not sure how to make the
hyperstack, or do I need to use it at all?

Any other comments for measuring PSF is also appreicated.

Thanks,

Zhengyu
> g Global Research Center
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> ______________________________________________________________________
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>           Zhengyu Pang, Ph.D.
                        Bioanalytics and Protein Science Laboratory
                        Biosciences Global Technology Organization
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> Niskayuna, NY12309
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