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Besides complex networks of protein interactions, the life of the cell is also governed by biophysical parameters. The objective of this course is to disseminate practical knowledge concerning recent quantitative fluorescence microscopy methods aiming at characterizing biophysical properties of the living samples. In the present practical course, we wish to introduce recent imaging methods allowing participants to assess three essential biophysical components of the intracellular environment: the diffusion-reaction dynamics the physico-chemical state the mechanical tensions Each component will be assessed in living cells and tissues using complementary methods. Hands-on sessions will focus not only on data acquisition but also on the methods available to quantitatively analyze and model these data.