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Live Cell Analysis Under Shear Flow

In today's life science research environment, the usefulness and versatility of cellular analysis is growing rapidly.

by Fluxion Biosciences
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Problem: In today’s life science research environment, the usefulness and versatility of cellular analysis is growing rapidly. While the evolution of research tools has enabled the generation of more data and increasingly detailed study of cellular function, static, plate-based assays have not been able to closely mimic conditions in the human body. Traditionally this has required moving the assays into in vivo studies in animal models, which is time consuming and expensive.

Solution: Fluxion Biosciences has developed research tools that bridge the gap between in vitro and in vivo, allowing advanced cellular study and minimizing the volume of animal studies. The foundation for these systems is cutting edge microfluidic technology, which can control shear flow in individual microplate wells to emulate in vivo conditions of the microvasculature and other physiological systems. Many processes behave differently under flow versus static conditions. Shear flow is important in vascular, immune and cardiovascular systems.

BioFlux well plate contains up to 24 individual flow cells, which are fully enclosed microfluidic channels for running flow-based experiments. The flow cells are arrayed on the bottom of an SBS-standard well plate with a conduit from the well to the microfluidic path. This enables direct introduction of reagents into the microfluidic path in the well, rather than through tubing or an off-chip reservoir. An airtight interface is then placed on the plate with a series of pneumatic connections to each of the wells. Flow is initiated using a pneumatic controller to move liquid through a flow channel thereby creating shear force within the physiological range. Shear values can also be rapidly switched on the fly, enabling complex flow protocols, such as cyclical waveforms.

BioFlux Plate channels viewed from beneath the well plates. Four wells are shown. Microfluidic flow cells are integrated into the bottom of the plate. Each fluidic channel runs between pairs of wells and has a control viewing window for observation.

Many immunology and cancer applications seek information on cell adhesion, migration and transmigration, which often occur under physiological, shear flow within the blood vessels. Live-cell assays on the BioFlux platform can provide data on growth of endothelial cell layers, rolling velocity measurements, cell adhesion counts, transmigration and high-content imaging. Small molecule assays can also be performed under flow conditions using the BioFlux, for increased physiological relevance.

In microbiology, the BioFlux platform can enhance the study of microbial communities, also called biofilms, many of which grow only in the presence of shear flow. Historical flow systems used to study biofilms have lacked the necessary throughput. Fluxion’s Well Plate Microfluidics (WPM™) can increase the throughput by as much as two orders of magnitude while the presence of controlled shear flow mimics conditions at the sites of biofilm infection. Anti-microbial efficacy and dose-response assays are also more effective under flow conditions and with WPM can be utilized as a cost-effective secondary screen.

Beyond immunology and microbiology, the BioFlux shear flow system has applications in stem cell research, neuronal cells and other specialized cell cultures. Further expansion of the system will include higher density well plates, alternate fluidic configurations for greater experimental flexibility and alternate detection modalities.

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