I have a number of cells transfected and labelled under different conditions, unfortunately on many the background from the staining is quite high.
In order to compare the fluorescence intensity, I want to take the ratio of the object (cell) integrated density to that of the background (or other way round).
I Plan to do this by
- segmenting the image
- measure integrated density of the cell
- inverse the selection by (Edit -> Selection -> Create selection and then Make Inverse)
- measure integrated density of the background
- ratio the values from 2 and 4 for each cell to see the background-corrected labelling for each condition
And because I am learning programming now, I want to make a macro for this to (in batch mode as well). I couldn’t find anything exactly like this so my questions to the more experienced image analysts here are:
- Has anyone encountered a macro doing exactly this (in which case I won’t waste my time and just study the macro instead)?
- Does it make sense what I want to do or am I over-complicating it and there is a simpler already established way to do the same thing?