We present here an innovative experimental methodology for the quantitative investigation of chemotaxis in vitro by live
imaging of cell movement in a reconstituted three-dimensional collagen gel. A well-defined chemoattractant gradient is
generated by means of a novel direct viewing chamber having two compartments (separated by a membrane), one containing
the chemoattractant solution, the other the cell-seeded collagen gel matrix. Cell migration is observed by means
of a time-lapse motorized video-microscopy workstation equipped with an incubating system and quantified by image
analysis techniques. Experimental results on three different cell lines (Jurkat, fibroblasts, and lymphocytes) are presented
for the isotropic control case (no chemoattractant) and in presence of a concentration gradient. Cell motility
data are in line with the concentration profile, both theoretically calculated from Fick’s law and experimentally measured
by epifluorescence microscopy. In particular, a transient peak in cell response was found, possibly due to cell
membrane receptor saturation