Abstract
Determining the structure and the internal dynamics of tissues is essential to understand their functional organization. Microscopy allows for monitoring positions and trajectories of every single cell. Those data are useful to extract statistical observables, such as intercellular distance, tissue symmetry and anisotropy, and cell motility. However, this procedure requires a large and supervised computational effort. In addition, due to the large cross-section of cells, the light scattering limits the use of microscopy to relatively thin samples. As an alternative approach, we propose to take advantage of light scattering and to analyze the dynamical diffraction pattern produced by a living tissue illuminated with coherent light. In this article, we illustrate with a few examples that supra-cellular structures produce an exploitable diffraction signal. From the diffraction signal, we deduce the mean distance between cells, the anisotropy of the supra-cellular organization and, from its fluctuations, the mean speed of moving cells. This easy to implement technique considerably reduces analysis time, allowing real time monitoring.
Highlights
In this article, we describe a non-imaging approach to investigate the structure and dynamics of living multicellular structures
The proposed method is based on the analysis of the dynamic speckle pattern produced by a set of cells illuminated with coherent light
This part was suppressed by applying a virtual beamstop at the center of the detector [black disk in the center of figure 2 (b)].Background light is substracted and vignetting effects are Structure and dynamics of multicellular assemblies measured by coherent light scattering8 corrected [25]
Summary
We describe a non-imaging approach to investigate the structure and dynamics of living multicellular structures. The proposed method is based on the analysis of the dynamic speckle pattern produced by a set of cells illuminated with coherent light. Video microscopy allows to monitor such complex cascade of events in great details and the analysis of the acquired time sequence of images provides accurate measures of position, shape and trajectory of each cell inside the tissue, from birth to death [3]. We propose to directly measure these observables by analyzing the dynamical speckle pattern produced by a living tissue illuminated with a collimated laser beam. The drawback of this self-averaging method is to lose the cell-to-cell heterogeneity, which might be crucial in certain circumstances
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