Fig. 1. Experimental set-up and alignment determination. (A) Design features of radial interstitial flow tissue culture chamber. The chamber is made of porous polyethylene and surface-modified glass materials to anchor the ECM and allow direct visualization. (B) Algorithm for image quantification of alignment and orientation. The confocal image (i) is modified (ii) to remove edge effects. (iii) A FFT transformation is performed to obtain a power spectrum from which (iv) an intensity frequency histogram is plotted and an alignment index (=(/( + ))/(/( + ))_ideal) and the peak angle (_peak) extracted.