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1.
ACS Sens ; 6(9): 3242-3252, 2021 09 24.
Article in English | MEDLINE | ID: mdl-34467761

ABSTRACT

The emergence of epigenetic gene regulation and its role in disease have motivated a growing field of epigenetic diagnostics for risk assessment and screening. In particular, irregular cytosine DNA base methylation has been implicated in several diseases, yet the methods for detecting these epigenetic marks are limited to lengthy protocols requiring bulky and costly equipment. We demonstrate a simple workflow for detecting methylated CpG dinucleotides in synthetic and genomic DNA samples using methylation-sensitive restriction enzyme digestion followed by loop-mediated isothermal amplification. We additionally demonstrate a cost-effective mobile fluorescence reader comprising a light-emitting diode bundle, a mirror, and optical fibers to transduce fluorescence signals associated with DNA amplification. The workflow can be performed in approximately 1 h, requiring only a simple heat source, and can therefore provide a foundation for distributable point-of-care testing of DNA methylation levels.


Subject(s)
Nucleic Acids , Fluorescence , Methylation , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques
2.
Sci Rep ; 7(1): 10802, 2017 09 07.
Article in English | MEDLINE | ID: mdl-28883551

ABSTRACT

Euglena gracilis (E. gracilis) has been proposed as one of the most attractive microalgae species for biodiesel and biomass production, which exhibits a number of shapes, such as spherical, spindle-shaped, and elongated. Shape is an important biomarker for E. gracilis, serving as an indicator of biological clock status, photosynthetic and respiratory capacity, cell-cycle phase, and environmental condition. The ability to prepare E. gracilis of uniform shape at high purities has significant implications for various applications in biological research and industrial processes. Here, we adopt a label-free, high-throughput, and continuous technique utilizing inertial microfluidics to separate E. gracilis by a key shape parameter-cell aspect ratio (AR). The microfluidic device consists of a straight rectangular microchannel, a gradually expanding region, and five outlets with fluidic resistors, allowing for inertial focusing and ordering, enhancement of the differences in cell lateral positions, and accurate separation, respectively. By making use of the shape-activated differences in lateral inertial focusing dynamic equilibrium positions, E. gracilis with different ARs ranging from 1 to 7 are directed to different outlets.


Subject(s)
Cell Separation/methods , Cell Shape , Euglena gracilis/cytology , Euglena gracilis/isolation & purification , Microfluidics/methods , Euglena gracilis/classification , Microfluidics/instrumentation
3.
Lab Chip ; 14(13): 2212-6, 2014 Jul 07.
Article in English | MEDLINE | ID: mdl-24852415

ABSTRACT

Here we highlight emerging technologies in the synthesis, handling, and application of encoded microparticles for multiplexed assays. Traditionally, in drug discovery and life sciences research, multiple reactions will be conducted in parallel using microwell plate formats or microfluidic implementations, in which volumes are confined and reactions annotated by knowledge of what reagents were added to each volume. Microparticle-based information carriers provide an alternative approach to performing such multiplexed reactions, in which reactions and events are instead annotated with unique codes associated with the solid-phase particle. One challenge has been in creating a unique and large enough code set that is also easily readout, and we highlight two approaches that have brought orthogonal optical tagging techniques to bear. Another challenge has been that in such approaches, reactions have usually been confined to the surface of, or within the bulk of the specifically-tagged particle. We also highlight a creative approach and strategy for multiplexing - called "partipetting"- in which the coded particle can be a carrier of a unique fluid reagent.


Subject(s)
Biological Assay , Microfluidic Analytical Techniques , Animals , Biological Assay/instrumentation , Biological Assay/methods , Humans , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods , Optics and Photonics/instrumentation , Optics and Photonics/methods
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