Measure the distribution of fluorescence

[dayani]

Hi,

I would like to measure and quantify the distribution of my fluorescence tagged protein of interest in the cell. To compare it upon knockdown and over-expression of other proteins, which affect its distribution in the cell, hence leading to a clustering phenotype.

Does anyone know how to measure and quantify it with the ImageJ software? Do I need to get a specific PlugIn for that?

Can anyone help please- I would really appreciate it.

Thanks in advance.

Dayani

[sombreroduck]

Hi,
I don't know anything about proteins and cells, but do you mean that you want to measure how large of the area of an image shows fluorescence? (which presumably is a distinct colour?)

If so, then I would normally ramp up the contrast so that it stands out distinct from the rest of the image, and then run a histogram. ImageJ has a functions Analyze->Histogram, though to be honest I normally do this type of analysis in Gimp or Photoshop because their histogram function tells one directly how many % of the image is of the specified colour-range.

Does this help?
R
(PS I find it easier working in greyscale, since then I only have the one channel)