Quantitative and multiplex microRNA assays from unprocessed cells in isolated nanoliter well arrays

Lab on a Chip
Augusto M TentoriPatrick S Doyle

Abstract

MicroRNAs (miRNAs) have recently emerged as promising biomarkers for the profiling of diseases. Translation of miRNA biomarkers to clinical practice, however, remains a challenge due to the lack of analysis platforms for sensitive, quantitative, and multiplex miRNA assays that have simple and robust workflows suitable for translation. The platform we present here utilizes functionalized hydrogel posts contained within isolated nanoliter well reactors for quantitative and multiplex assays directly from unprocessed cell samples without the need of prior nucleic acid extraction. Simultaneous reactor isolation and delivery of miRNA extraction reagents is achieved by sealing an array of wells containing the functionalized hydrogel posts and cells against another array of wells containing lysis and extraction reagents. The nanoliter well array platform features >100× better sensitivity compared to previous technology utilizing hydrogel particles without relying on signal amplification and enables >100 parallel assays in a single device. These advances provided by this platform lay the groundwork for translatable and robust analysis technologies for miRNA expression profiling in samples with small populations of cells and in precious,...Continue Reading

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Citations

Dec 11, 2019·International Journal of Molecular Sciences·Wen Cai Zhang
May 19, 2020·Microsystems & Nanoengineering·Maxwell B NagarajanPatrick S Doyle
Dec 3, 2020·APL Bioengineering·Alden C Moss, Amy E Herr

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Methods Mentioned

BETA
nucleic acid extraction
RNA-seq
biosensors
biopsies
fluorescence imaging
Fluorescence

Software Mentioned

MATLAB
ImageJ
Andor Solis

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