High-throughput microfluidic single-cell RT-qPCR
Adam K. White,Michael VanInsberghe,Oleh Petriv,Mani Hamidi,Darek Sikorski,Marco A. Marra,James M. Piret,Samuel Aparicio,Carl L. Hansen +8 more
TLDR
This work presents a fully integrated microfluidic device capable of performing high-precision RT-qPCR measurements of gene expression from hundreds of single cells per run, and shows that nanoliter volume processing reduced measurement noise, increased sensitivity, and provided single nucleotide specificity.Abstract:
A long-sought milestone in microfluidics research has been the development of integrated technology for scalable analysis of transcription in single cells Here we present a fully integrated microfluidic device capable of performing high-precision RT-qPCR measurements of gene expression from hundreds of single cells per run Our device executes all steps of single-cell processing, including cell capture, cell lysis, reverse transcription, and quantitative PCR In addition to higher throughput and reduced cost, we show that nanoliter volume processing reduced measurement noise, increased sensitivity, and provided single nucleotide specificity We apply this technology to 3,300 single-cell measurements of (i) miRNA expression in K562 cells, (ii) coregulation of a miRNA and one of its target transcripts during differentiation in embryonic stem cells, and (iii) single nucleotide variant detection in primary lobular breast cancer cells The core functionality established here provides the foundation from which a variety of on-chip single-cell transcription analyses will be developedread more
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Dissertation
Heat, Fluid, and Sample Control in Point-of-Care Diagnostics
TL;DR: Heat, Fluid, and Sample Control in Point-of-Care Diagnostics examines the role of heat, fluid, and sample control in point- of-Care diagnosis.
Journal ArticleDOI
Cutting-Edge Microfabricated Biomedical Tools for Human Pluripotent Stem Cell Research
TL;DR: This review describes advanced microfabricated tools for studying the mechanisms of in vivo stem cell regulation and precise manipulation of stem cells, notably hPSCs, and concerns of using such tools for stem cell research.
Journal ArticleDOI
A microfluidic chip for the versatile chemical analysis of single cells.
TL;DR: A microfluidic device that enables the quantitative determination of intracellular biomolecules in multiple single cells in parallel, and many microchambers can be arranged in an array format, allowing the analysis of many cells at once, given that suitable optical instruments are used for monitoring.
Journal ArticleDOI
Quantitative Detection and Real-Time Monitoring of Endogenous mRNA at the Single Live Cell Level Using a Ratiometric Molecular Beacon.
TL;DR: The designed RMB could detect the Hsp27 mRNA with high signal-to-noise ratio and sensitivity as well as excellent specificity and anti-degradation capability proved in vitro and in live cells, demonstrating that the proposed RMB can be a potential quantitative endogenous mRNA detection tool especially at a single live cell level.
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Advances in biotechnology and linking outputs to variation in complex traits: Plant and Animal Genome meeting January 2012
TL;DR: The Plant and Animal Genome meeting in January 2012 provided insights into the advances in plant, animal, and microbe genome studies particularly as they impact on the authors' understanding of complex biological systems.
References
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Journal ArticleDOI
MicroRNAs: Genomics, Biogenesis, Mechanism, and Function
TL;DR: Although they escaped notice until relatively recently, miRNAs comprise one of the more abundant classes of gene regulatory molecules in multicellular organisms and likely influence the output of many protein-coding genes.
Journal ArticleDOI
MicroRNA expression profiles classify human cancers
Jun Lu,Gad Getz,Eric A. Miska,Eric A. Miska,Ezequiel Alvarez-Saavedra,Justin Lamb,David Peck,Alejandro Sweet-Cordero,Alejandro Sweet-Cordero,Benjamin L. Ebert,Benjamin L. Ebert,Raymond H. Mak,Raymond H. Mak,Adolfo A. Ferrando,James R. Downing,Tyler Jacks,H. Robert Horvitz,H. Robert Horvitz,Todd R. Golub,Todd R. Golub,Todd R. Golub +20 more
TL;DR: A new, bead-based flow cytometric miRNA expression profiling method is used to present a systematic expression analysis of 217 mammalian miRNAs from 334 samples, including multiple human cancers, and finds the miRNA profiles are surprisingly informative, reflecting the developmental lineage and differentiation state of the tumours.
Journal ArticleDOI
Real-time quantification of microRNAs by stem–loop RT–PCR
Caifu Chen,Dana Ridzon,Adam Broomer,Zhaohui Zhou,Danny H. Lee,Julie T. Nguyen,Maura Barbisin,Nan Lan Xu,Vikram R. Mahuvakar,Mark R. Andersen,Kaiqin Lao,Kenneth J. Livak,Karl J. Guegler +12 more
TL;DR: A novel microRNA quantification method has been developed using stem–loop RT followed by TaqMan PCR analysis, which enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases.
Journal ArticleDOI
Monolithic microfabricated valves and pumps by multilayer soft lithography
TL;DR: An extension to the soft lithography paradigm, multilayersoft lithography, with which devices consisting of multiple layers may be fabricated from soft materials is described, to build active microfluidic systems containing on-off valves, switching valves, and pumps entirely out of elastomer.
Journal ArticleDOI
A mammalian microRNA expression atlas based on small RNA library sequencing.
Pablo Landgraf,Mirabela Rusu,Robert L. Sheridan,Alain Sewer,Alain Sewer,Nicola Iovino,Alexei A. Aravin,Sébastien Pfeffer,Amanda J. Rice,Alice O. Kamphorst,Markus Landthaler,Carolina Lin,Nicholas D. Socci,Leandro C. Hermida,Valerio Fulci,Sabina Chiaretti,Robin Foà,Julia Schliwka,Uta Fuchs,Astrid Novosel,Roman-Ulrich Müller,Roman-Ulrich Müller,Bernhard Schermer,Ute Bissels,Jason M. Inman,Quang Phan,Minchen Chien,David B. Weir,Ruchi Choksi,Gabriella De Vita,Daniela Frezzetti,Hans Ingo Trompeter,Veit Hornung,Grace Teng,Gunther Hartmann,Miklós Palkovits,Roberto Di Lauro,Peter Wernet,Giuseppe Macino,Charles E. Rogler,James W. Nagle,Jingyue Ju,F. Nina Papavasiliou,Thomas Benzing,Peter Lichter,Wayne Tam,Michael J. Brownstein,Andreas Bosio,Arndt Borkhardt,James J. Russo,Chris Sander,Mihaela Zavolan,Mihaela Zavolan,Thomas Tuschl +53 more
TL;DR: A relatively small set of miRNAs, many of which are ubiquitously expressed, account for most of the differences in miRNA profiles between cell lineages and tissues.
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