ArticleAnalytical chemistry2014
Self-digitization microfluidic chip for absolute quantification of mRNA in single cells.
Article in Analytical chemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
24 citing papers in PubMed, 63 citations in OpenAlex.
- Digital PCR for Single-Cell Analysis.Biosensors · 2024Review
- Robust and rapid partitioning in thermoplastic.The Analyst · 2023Article
- Dielectrophoresis-Assisted Self-Digitization Chip for High-Efficiency Single-Cell Analysis.Methods in molecular biology (Clifton, N.J.) · 2023Article
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- A reinforced PDMS mold for hot embossing of cyclic olefin polymer in the fabrication of microfluidic chips.Lab on a chip · 2022Article
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- Microdroplet-based one-step RT-PCR for ultrahigh throughput single-cell multiplex gene expression analysis and rare cell detection.Scientific reports · 2021Article
- A Double-Deck Self-Digitization Microfluidic Chip for Digital PCR.Micromachines · 2020Article
- PCR past, present and future.BioTechniques · 2020Review
- Enhanced sample filling and discretization in thermoplastic 2D microwell arrays using asymmetric contact angles.Biomicrofluidics · 2020Article
- A microfluidic alternating-pull-push active digitization method for sample-loss-free digital PCR.Lab on a chip · 2019Article
- scMatch: a single-cell gene expression profile annotation tool using reference datasets.Bioinformatics (Oxford, England) · 2019Article
- Self-digitization chip for quantitative detection of human papillomavirus gene using digital LAMP.Lab on a chip · 2019Article
- Ultrasensitive quantification of tumor mRNAs in extracellular vesicles with an integrated microfluidic digital analysis chip.Lab on a chip · 2018Article
- SD-chip enabled quantitative detection of HIV RNA using digital nucleic acid sequence-based amplification (dNASBA).Lab on a chip · 2018Article
- A Self-Digitization Dielectrophoretic (SD-DEP) Chip for High-Efficiency Single-Cell Capture, On-Demand Compartmentalization, and Downstream Nucleic Acid Analysis.Angewandte Chemie (International ed. in English) · 2018Article
- dPCR: A Technology Review.Sensors (Basel, Switzerland) · 2018Review
- Ultrasensitive Quantification of Cytokine Proteins in Single Lymphocytes From Human Blood FollowingFrontiers in immunology · 2018Article
- Article
- Review
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
Quantification of mRNA in single cells provides direct insight into how intercellular heterogeneity plays a role in disease progression and outcomes. Quantitative polymerase chain reaction (qPCR), the current gold standard for evaluating gene expression, is insufficient for providing absolute measurement of single-cell mRNA transcript abundance. Challenges include difficulties in handling small sample volumes and the high variability in measurements. Microfluidic digital PCR provides far better sensitivity for minute quantities of genetic material, but the typical format of this assay does not allow for counting of the absolute number of mRNA transcripts samples taken from single cells. Furthermore, a large fraction of the sample is often lost during sample handling in microfluidic digital PCR. Here, we report the absolute quantification of single-cell mRNA transcripts by digital, one-step reverse transcription PCR in a simple microfluidic array device called the self-digitization (SD) chip. By performing the reverse transcription step in digitized volumes, we find that the assay exhibits a linear signal across a wide range of total RNA concentrations and agrees well with standard curve qPCR. The SD chip is found to digitize a high percentage (86.7%) of the sample for single-cell experiments. Moreover, quantification of transferrin receptor mRNA in single cells agrees well with single-molecule fluorescence in situ hybridization experiments. The SD platform for absolute quantification of single-cell mRNA can be optimized for other genes and may be useful as an independent control method for the validation of mRNA quantification techniques.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.