Measuring nuclear translocation of GFP tagged protein


#1

Dear CellProfiler Experts,

First at all: Great program! It already helped me dealing with dozens of problems and challenges.
At present, I try to measure the level of nuclear translocation of a GFP tagged protein (DAF-16::GFP in C. elegans). This protein is distributed in the cell in unstressed conditions (left picture) and accumulates in the nucleus after stress exposure (right picture). I would like to quantify the degree of accumulation. I have two questions about it:

  1. I tried to use the “MeasureGranularity” tool to quantify the translocation, but I do not know how to interpret the result; for example: I get the value “31.79” for “Granularity 1” and “0.57” for “Granularity 13”. Are the 1 and the 13 the radius of the respective granules? But what are the numbers ‘31.79’ and ‘0.57’ mean then? I thought this tool could maybe count the number of granules in a given range, but doesn’t look like?!

  2. Is there maybe a more suitable tool, which can show me how homogeneous the fluorescence is distributed in my primary object (the worm)? The decrease of homogeneity should be measureable in worms where GFP is mainly located in the nucleus, right?

Thank you very much for any help or idea!!
Nadine

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