Here we seek to expand on previous work which combined Quantitative Phase Imaging (QPI) with a molecularly sensitive Förster resonance energy transfer (FRET) construct by developing additional analysis techniques. This work seeks to further characterize the mechanotransduction of individual cells in response to stimulus through a novel QPI-guided cellular segmentation algorithm. The multimodal imaging instrument and analysis techniques are employed to examine cellular responses to hypo-osmotic stimulus by observing the calcium ion flux using a FRET based sensor coupled with a mapping of intracellular mass reorganization using QPI. The combined imaging modality enables a discrimination of cell response by localized region revealing distinct behavior between regions and relative to a control group.
