The Role of Cellular Environment in Dynamic Light Scattering

摘要

We have developed motility contrast imaging (MCI) as a coherence-domain volumetric imaging approach that uses subcellular dynamics as an endogenous imaging contrast agent of living tissue. Fluctuation spectroscopy analysis of dynamic light scattering (DLS) from 3-D tissue has identified functional frequency bands related to organelle transport, membrane undulations and cell shape change. In this paper, we track the behavior of dynamic light scattering as we bridge the gap between the two extremes of 2-D cell culture on the one hand, and 3-D tissue spheroids on the other. In a light backscattering geometry, we capture speckle from 2-D cell culture consisting of isolated cells or planar rafts of cells on cell-culture surfaces. DLS from that cell culture shows differences and lower sensitivity to intra-cellular dynamics compared with the 3-D tissue. The motility contrast is weak in this limit. As the cellular density increases to cover the surface, the motility contrast increases. As environmental perturbations or Pharmaceuticals are applied, the fluctuation spectral response becomes more dramatic as the dimensionality of the cellular aggregations increases. We show that changing optical thickness of the cellular-to-tissue targets usually causes characteristic frequency shifts in the spectrograms, while changing cellular dimensionality causes characteristic frequencies to be enhanced or suppressed.