Microparticle parking and isolation for highly sensitive microRNA detection
| Autor: | Jae Jung Kim, Patrick S. Doyle, Lynna Chen |
|---|---|
| Přispěvatelé: | Massachusetts Institute of Technology. Department of Chemical Engineering, Massachusetts Institute of Technology. Department of Biological Engineering |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
In situ Materials science Kinetics Microfluidics Biomedical Engineering Bioengineering Nanotechnology Biochemistry complex mixtures Sensitivity and Specificity Article 03 medical and health sciences Microparticle Particle Size Aqueous solution technology industry and agriculture Hydrogels General Chemistry Equipment Design RNA Probes Microfluidic Analytical Techniques Highly sensitive MicroRNAs 030104 developmental biology Self-healing hydrogels Particle size |
| Zdroj: | PMC |
| Popis: | Isolating small objects, such as particles, cells, and molecules, in individual aqueous droplets is useful for chemical and biological assays. We have developed a simple microfluidic platform to immobilize (park) microparticles at defined locations, and isolate particles in monodisperse droplets surrounded by immiscible oil. While conventional methods can only achieve stochastic encapsulation of objects within larger droplets, our in situ method ensures that a single particle is entrapped in a similar-sized droplet, with ∼95% yield for parking and isolation. This enables time-lapse studies of reactions in confined volumes and can be used to perform enzymatic amplification of a desired signal to improve the sensitivity of diagnostic assays. To demonstrate the utility of our technique, we perform highly sensitive, multiplexed microRNA detection by isolating encoded, functional hydrogel microparticles in small aqueous droplets. Non-fouling hydrogel microparticles are attractive for microRNA detection due to favorable capture kinetics. By encapsulating these particles in droplets and employing a generalizable enzyme amplification scheme, we demonstrate an order of magnitude improvement in detection sensitivity compared to a non-amplified assay. ©2017 The Royal Society of Chemistry. NIH-NCI Grant 5R21CA177393-02 |
| Databáze: | OpenAIRE |
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