Visualizing hypoxic modulation of beta cell secretions via a sensor augmented oxygen gradient.

One distinct advantage of microfluidic-based cell assays is their scalability for multiple concentrations or gradients. Microfluidic scaling can be extremely powerful when combining multiple parameters and modalities. Moreover, in situ stimulation and detection eliminates variability between individual bioassays. However, conventional microfluidics must combat diffusion, which limits the spatial distance and time for molecules traveling through microchannels. Here, we leveraged a multilayered microfluidic approach to integrate a novel oxygen gradient (0-20%) with an enhanced hydrogel sensor to study pancreatic beta cells. This enabled our microfluidics to achieve spatiotemporal detection that is difficult to achieve with traditional microfluidics. Using this device, we demonstrated the in situ detection of calcium, insulin, and ATP (adenosine triphosphate) in response to glucose and oxygen stimulation. Specifically, insulin was quantified at levels as low as 25 pg/mL using our imaging technique. Furthermore, by analyzing the spatial detection data dynamically over time, we uncovered a new relationship between oxygen and beta cell oscillations. We observed an optimum oxygen level between 10 and 12%, which is neither hypoxic nor normoxic in the conventional cell culture sense. These results provide evidence to support the current islet oscillator model. In future applications, this spatial microfluidic technique can be adapted for discrete protein detection in a robust platform to study numerous oxygen-dependent tissue dysfunctions.