Combining quantitative phase microscopy and laser-induced shockwave for the study of cell injury.

In this paper, we propose a new system for studying cellular injury. The system is a biophotonic work station that can generate Laser-Induced Shockwave (LIS) in the cell culture medium combined with a Quantitative Phase Microscope (QPM), enabling the real-time measurement of intracellular dynamics and quantitative changes in cellular thickness during the damage and recovery processes. In addition, the system is capable of Phase Contrast (PhC) and Differential Interference Contrast (DIC) microscopy. Our studies showed that QPM allows us to discern changes that otherwise would be unnoticeable or difficult to detect using phase or DIC imaging. As one application, this system enables the study of traumatic brain injury in vitro. Astrocytes are the most numerous cells in the central nervous system (CNS) and have been shown to play a role in the repair of damaged neuronal tissue. In this study, we use LIS to create a precise mechanical force in the culture medium at a controlled distance from astrocytes and measure the quantitative changes, in order of nanometers, in cell thickness. Experiments were performed in different cell culture media in order to evaluate the reproducibility of the experimental method.

Fluid Shear Stress Enhances the Phagocytic Response of Astrocytes.

Astrocytes respond to brain injury at a cellular level by the process of reactive astrogliosis, and are able to adjust their response according to the severity of the insult. Included in the reactive response is the process of phagocytosis, where astrocytes clean up surrounding cellular debris from damaged cells. In this study, we observe the process of phagocytosis by primary cortical astrocytes in the presence of media flow across the apical surface of the cells. Both static and cells under flow conditions respond consistently via phagocytosis of laser-induced cellular debris. We found that astrocytes exposed to shear flow initiate phagocytosis at a consistently faster rate than cells observed under static conditions. Shear forces created by laminar flow were analyzed as well as the flow fields created around astrocyte cells. Results suggest astrocyte phagocytosis is a mechanosensitive response, thus revealing the potential to enhance astrocyte phagocytic cleanup of damaged nervous tissue.