http://imagej.273.s1.nabble.com/Changes-in-gfp-fluorence-in-nucleus-tp5017628p5017640.html
> Hi everyone!
>
> I have the following problem:
>
> I have a gfp tagged protein that localizes in damaged dna (i
> microirradiate a part of the nucleus with a laser and my protein moves
> there. In untreated cells the nucleus remain completely black).
>
> I also have different mutants for this protein that seems to be affecting
> how this protein behave (looks like the mutants enter the nucleus but don't
> quite localize at the damaged sites like the WT protein).
>
> I have different time points after the irradiation and i was approachibg
> the situation like this:
>
> 1) Select a roi for the nucleus and another for the irradiated zone.
> 2) On the photos from the green channel:
> -I do a threshold correction until the nucleus at time 0 is black.
> -calculate the area and the integrated density for both rois.
> - Move to the next time point. Apply the same value of threshold
> obtained at time 0 and the do the same calculations.
> -For each cell i calculate the intDen/area = Integrated density / area
> for the entire nucleus, the irradiated zone and the non irradiated zone
> (nucleus minus irradiated zone).
> -Then calculate a ratio of the previous parameter versus the time 0 to
> see the changes over time.
>
> My idea was to compare how the fluorescence changes over time in the non
> irradiated zone and the irradiated zones among the different mutants i have
> but i'm not sure this is the right way to do it. I have a lot of cells and
> mutants and before doing a lot of work i would like to know if this is the
> right way to do it.
>
> Many thanks in advance for your answers,
>
> Gonzalo
>
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