Calculating the individual spot intensities from an array of spots in an image (like micro array image)

[Adriot]

HI..

I am trying to calculating the average intensity of each individual spot in a micro array ( Which consists of array of circular spots).

is there any possible solution to find out thier average intensities and other parameters after calculating the centroid of each spot or even with out calculating all the centriod

Please find the image attached micro array image for ref

Thanks in advance

[Henrik_Volkers]

Shouldn't be too hard with the vision package ...

(I haven't used it yet, however after 3a of programming optical vision systems )

So without: I assume you have an 2D array of values: (and the spots don't merge )  

Do you have them roughly alined?  Say you have horizontal lines  and vertical columns in your array that are all (nearly) zero?

1. You can find them by looking at the total (sum) of the columns and lines  and look for maxima and minima (zero?) 

2. Do a coarse separation (get a sub array)  from every spot area (position and size calculated by step 1)

3. Totalize your columns and lines: the maxima give the position. You can do a functional fit do get sub pixel resolution.

4. The minima should be at the borders, if not your 'stamp out'  didn't only catch one spot. (test if not use result of 3 and go to step 2)  

5. The total of your subarray will be the intensity.