A review of chemical gradient systems for cell analysis.

Microfluidic spatial and temporal gradient generators have played an important role in many biological assays such as in the analysis of wound healing, inflammation, and cancer metastasis. Chemical gradient systems can also be applied to other fields such as drug design, chemical synthesis, chemotaxis, etc. Microfluidic systems are particularly amenable to gradient formation, as the length scales used in chips enable fluid processes that cannot be conducted in bulk scale. In this review we discuss new microfluidic devices for gradient generation and applications of those systems in cell analysis.

On-chip gradient generation in 256 microfluidic cell cultures: simulation and experimental validation.

A microfluidic diffusion diluter was used to create a stable concentration gradient for dose response studies. The microfluidic diffusion diluter used in this study consisted of 128 culture chambers on each side of the main fluidic channel. A calibration method was used to find unknown concentrations with 12% error. Flow rate dependent studies showed that changing the flow rates generated different gradient patterns. Mathematical simulations using COMSOL Multi-physics were performed to validate the experimental data. The experimental data obtained for the flow rate studies agreed with the simulation results. Cells could be loaded into culture chambers using vacuum actuation and cultured for long times under low shear stress. Decreasing the size of the culture chambers resulted in faster gradient formation (20 min). Mass transport into the side channels of the microfluidic diffusion diluter used in this study is an important factor in creating the gradient using diffusional mixing as a function of the distance. To demonstrate the device's utility, an H2O2 gradient was generated while culturing Ramos cells. Cell viability was assayed in the 256 culture chambers, each at a discrete H2O2 concentration. As expected, the cell viability for the high concentration side channels increased (by injecting H2O2) whereas the cell viability in the low concentration side channels decreased along the chip due to diffusional mixing as a function of distance. COMSOL simulations were used to identify the effective concentration of H2O2 for cell viability in each side chamber at 45 min. The gradient effects were confirmed using traditional H2O2 culture experiments. Viability of cells in the microfluidic device under gradient conditions showed a linear relationship with the viability of the traditional culture experiment. Development of the microfluidic device used in this study could be used to study hundreds of concentrations of a compound in a single experiment.

Cavity-enhanced absorption measurements across broad absorbance and reflectivity ranges.

Cavity-enhanced spectrometry constitutes an important and highly sensitive technique for absorbance measurements. The current practice generally involves very high reflectivity mirrors and hence intense light sources (typically lasers) to have enough light transmitted. Available theory describes the situation only for high-finesse cavities (high-reflectance mirrors) and generally for systems with very low absorbances. We develop the general expression for absorbance regardless of mirror reflectivity or the absorbance and show that in the limit of high reflectivities and low absorbances it predicts the same numerical values as that derived by O'Keefe (Chem. Phys. Lett. 1998, 293, 331-336; Chem. Phys. Lett. 1999, 307, 343-349). Signal to noise in any photometric system is also dependent on the amount of light reaching the detector because of shot noise limitations. We show that a small aperture in the entrance mirror greatly improves light throughput without significant departure from the theoretically predicted amplification of absorbance; such simple modifications result in real improvement of detection limits, even with mirrors of modest reflectivity and inexpensive detectors. This allows the merits of cavity enhancement measurements to be demonstrated for pedagogic purposes.

A computational model of fibroblast and macrophage spatial/temporal dynamics in foreign body reactions.

The implantation of medical devices often triggers several immune responses, one kind of which is categorized as foreign body reactions. It is well established that macrophages and many other cells participate in the complex processes of foreign body reactions, and cause severe inflammations and fibrotic capsule formation in surrounding tissues. However, the detailed mechanisms of macrophage responses, recruitment and activation, in foreign body reactions are not totally understood. In the meantime, mathematical models have been proposed to systematically decipher the behavior of this complex system of multiple cells, proteins and biochemical processes in wound healing responses. Based on these early works, this study introduces a mathematical model in two spatial dimensions to investigate the transient behavior of macrophages, fibroblasts and their interactions during the formation of fibrotic tissue. We find that the simulation results are consistent with the experimental observations. These findings support that the model can reveal quantitative insights for studying foreign body reaction processes.