Estimation of available epinephrine dose in expired and discolored autoinjectors via quantitative smartphone imaging.

Epinephrine autoinjectors (EAIs) are important first aid medications for treating anaphylaxis. A 10-fold price increase over the past 12 years and evidence that expired EAIs may still contain significant doses of available epinephrine have motivated interest in the efficacy of expired EAIs as treatments of last resort. Degradation of expired EAIs, which can be caused by improper storage conditions, results in various degrees of discoloration of the epinephrine solution. Previous studies have determined that significant epinephrine remains available in expired EAIs, but these have only considered EAIs that show no discoloration. Here, we investigate the potential for colorimetric estimation of available epinephrine dose based on the degree of discoloration in expired EAIs. The correlation of available epinephrine dose and time since expiration date was poor (r = - 0.37), as determined by an industry standard UHPLC protocol. Visible absorbance of the samples integrated across the range 430-475 nm correlated well with available epinephrine dose (r = - 0.71). This wavelength corresponds to the blue channel of a typical smartphone camera Bayer filter. Smartphone camera images of the EAI solutions in various illumination conditions were analyzed to assign color indices representing the degree of discoloration. Color index of the samples showed similar correlation (|r| > 0.7) with available epinephrine dose as that of visible spectrophotometry. Smartphone imaging colorimetry is proposed as a potential point-of-use epinephrine dose estimator for expired and degraded EAIs. Graphical abstract.

Total Internal Reflection Transient Absorption Microscopy: An Online Detection Method for Microfluidics.

Microreactors have garnered widespread attention for their tunability and precise control of synthetic parameters to efficiently produce target species. Despite associated advances, a lack of online detection and optimization methods has stalled the progression of microfluidic reactors. Here we employ and characterize a total internal reflection transient absorption microscopy (TIRTAM) instrument to image excited state dynamics on a continuous flow device. The experiments presented demonstrate the capability to discriminate between different chromophores as well as in differentiating the effects of local chemical environments that a chromophore experiences. This work presents the first such online transient absorption measurements and provides a new direction for the advancement and optimization of chemical reactions in microfluidic devices.

A Scalable, Modular Degasser for Passive In-Line Removal of Bubbles from Biomicrofluidic Devices.

Bubbles are a common cause of microfluidic malfunction, as they can perturb the fluid flow within the micro-sized features of a device. Since gas bubbles form easily within warm cell culture reagents, degassing is often necessary for biomicrofluidic systems. However, fabrication of a microscale degasser that can be used modularly with pre-existing chips may be cumbersome or challenging, especially for labs not equipped for traditional microfabrication, and current commercial options can be expensive. Here, we address the need for an affordable, accessible bubble trap that can be used in-line for continuous perfusion of organs-on-chip and other microfluidic cultures. We converted a previously described, manually fabricated PDMS degasser to allow scaled up, reproducible manufacturing by commercial machining or fused deposition modeling (FDM) 3D printing. After optimization, the machined and 3D printed degassers were found to be stable for >2 weeks under constant perfusion, without leaks. With a ~140 µL chamber volume, trapping capacity was extrapolated to allow for ~5-20 weeks of degassing depending on the rate of bubble formation. The degassers were biocompatible for use with cell culture, and they successfully prevented bubbles from reaching a downstream microfluidic device. Both degasser materials showed little to no leaching. The machined degasser did not absorb reagents, while the FDM printed degasser absorbed a small amount, and both maintained fluidic integrity from 1 µL/min to >1 mL/min of pressure-driven flow. Thus, these degassers can be fabricated in bulk and allow for long-term, efficient bubble removal in a simple microfluidic perfusion set-up.

A microfluidic bubble perfusion device for brain slice culture.

Ex vivo brain slice cultures are utilized as analytical models for studying neurophysiology. Common approaches to maintaining slice cultures include roller tube and membrane interface techniques. The rise of organ-on-chip technologies has demonstrated the value of microfluidic perfusion culture systems for sampling and analysis of complex biology under well-controlled in vitro or ex vivo conditions. A number of approaches to microfluidic brain slice culture have been developed, however these typically involve complex design, fabrication, or operational parameters in order to meet the high oxygen demands of brain slices. Here, we present proof-of-principle for a novel approach to microfluidic brain slice culture. In this system, which we term a microfluidic bubble perfusion device, principles of droplet microfluidics were employed to generate droplets of perfusion media dispersed between bubbles of carbogen gas, and brain tissue slices were perfused with the resulting monodispersed droplets and bubbles. The challenge of tissue immobilization in the flow system was addressed using a two-part cytocompatible carbohydrate-based tissue adhesive. Best practices are discussed for perfusion chamber designs that maintain segmented flow throughout the course of perfusion. Control of droplet and bubble volumes was possible across the range of ca. 4-15 µL, bubble generation frequency was well controlled in the range ca. 1-7 bubbles per min, and bubble duty cycle was well controlled across the range ca. 20-80%. Murine hypothalamic tissue slices containing the suprachiasmatic nuclei were successfully maintained for durations of 8-10 hours, with tissue remaining viable for the duration of perfusion as assessed by Ca2+ imaging and propidium iodide (PI) staining.