Stationary nanoliter droplet array with a substrate of choice for single adherent/nonadherent cell incubation and analysis

Microfluidic water-in-oil droplets that serve as separate, chemically isolated compartments can be applied for single-cell analysis; however, to investigate encapsulated cells effectively over prolonged time periods, an array of droplets must remain stationary on a versatile substrate for optimal cell compatibility. We present here a platform of unique geometry and substrate versatility that generates a stationary nanodroplet array by using wells branching off a main microfluidic channel. These droplets are confined by multiple sides of a nanowell and are in direct contact with a biocompatible substrate of choice. The device is operated by a unique and reversed loading procedure that eliminates the need for fine pressure control or external tubing. Fluorocarbon oil isolates the droplets and provides soluble oxygen for the cells. By using this approach, the metabolic activity of single adherent cells was monitored continuously over time, and the concentration of viable pathogens in blood-derived samples was determined directly by measuring the number of colony-formed droplets. The method is simple to operate, requires a few microliters of reagent volume, is portable, is reusable, and allows for cell retrieval. This technology may be particularly useful for multiplexed assays for which prolonged and simultaneous visual inspection of many isolated single adherent or nonadherent cells is required.clos

A fidget spinner for the point-of-care diagnosis of urinary tract infection

The point-of-care detection of pathogens in biological samples in resource-limited settings should be inexpensive, rapid, portable, simple and accurate. Here, we describe a custom-made fidget spinner that rapidly concentrates pathogens in 1-ml samples of undiluted urine by more than 100-fold for the on-device colorimetric detection of bacterial load and pathogen identification. In Tiruchirappalli, India, the device enabled the on-site detection of infection with the naked eye within 50 min in urine samples from 39 patients suspected of having a urinary tract infection. We also show that, in 30 clinical samples of urinary tract infection, the device can be used to perform an antimicrobial susceptibility test for the antimicrobial drugs ciprofloxacin and cefazolin within 120 min. The fidget spinner could be used in low-resource settings as an inexpensive handheld point-of-care device for the rapid concentration and detection of pathogens in urine samples. A custom-made fidget spinner rapidly concentrates pathogens in 1-ml samples of undiluted urine by more than 100-fold for the on-device colorimetric detection of bacterial load and pathogen identification

Fabrication and Characterization of Autonomously Self-Healable and Stretchable Soft Microfluidics

In this paper, a novel self-healable and stretchable microfluidics system for next generation wearable lab-on-a-chip is presented. An imine-based precursor with various metal sources (Co(II), Fe(II), and Zn(II)) is used for the development of an intrinsically autonomous self-healing microfluidic device. Microfluidics fabrication is performed on the self-healing substrate layer using a mold transfer method. The mechanical properties of the resulting layer are evaluated using tensile strain pull testing. Microfluidic characteristics including fluid flow, wettability, leak, and fluorescence compatibility are investigated to understand its performance in classical microfluidic applications. The new microfluidic devices are also characterized using scanning-electron microscopy to evaluate the mold transfer capability. The self-healing microfluidics and the corresponding detailed fluidic characterization presented in this paper will open new opportunities for microfluidic lab on a chip development for various applications, especially in wearable electronics

Static droplet array for culturing single live adherent cells in an isolated chemical microenvironment

We present here a new method to easily and reliably generate an array of hundreds of dispersed nanoliter-volume semi-droplets for single-cells culture and analysis. The liquid segmentation step occurs directly in indexed traps by a tweezer-like mechanism and is stabilized by spatial confinement. Unlike common droplet-based techniques, the semi-droplet wets its surrounding trap walls thus supporting the culturing of both adherent and non-adherent cells. To eliminate cross-droplet cell migration and chemical cross-talk each semi-droplet is separated from a nearby trap by an ∼80 pL air plug. The overall setup and injection procedure takes less than 10 minutes, is insensitive to fabrication defects and supports cell recovery for downstream analysis. The method offers a new approach to easily capture, image and culture single cells in a chemically isolated microenvironment as a preliminary step towards high-throughput single-cell assays