Journal
LAB ON A CHIP
Volume 20, Issue 11, Pages 1939-1946Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c9lc01226e
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Funding
- City University of Hong Kong [9610467]
- National Research Foundation Singapore, Synthetic Biology Research Program (NRF, SBP) [R-397-000-276-281, R-397-000-323-592]
- National Research Foundation Singapore, Competitive Research Programme (NRF, CRP)
- National Medical Research Council Singapore, Open Fund - Individual Research Grant (NMRC, OFIRG) [R-397-000-289-213]
- Ministry of Education (MOE) Singapore, Tier-2 [R-397-000-271-112, R-279-000-501-112]
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Cell nucleocytoplasmic profiles of microRNAs (miRNAs) are critical to determining a single cell's essential functionalities, such as cellular transcription, nucleus export and degradation, which gives a comprehensive view of cellular processes. Despite the importance of addressing nucleocytoplasmic heterogeneity, the challenge of high-throughput screening remains. Although a droplet-based approach was developed for single-cell miRNA assays, the challenge of quantifying miRNA with high sensitivity to indicate nucleocytoplasmic heterogeneity remains. In this study, a nanoplasmon-enhanced droplet screening platform was developed to quantify single-cell nucleocytoplasmic heterogeneity with the high sensitivity of 0.1 nM. Droplet screening and multiplexed plasmonic assays are synergistic: droplet screening is used to isolate single cells for high-throughput screening, while enhanced nanoplasmonic assays are conducted to precisely determine different types of miRNAs, addressing the cell nucleocytoplasmic profile. Here, two nucleic acid-functionalized plasmonic nanosensors, silver nanoparticles functionalized with designed sequences to target miRNAs, are synthesized. After the targets are bound, competitive formation of sensor-target hybrids interferes with plasmonic coupling between the nanoparticles, decreasing a fluorescence signal and thus enabling high-sensitivity single-cell miRNA quantification. Using the fluorescence signal change as a readout allows continuous-flow measurement to provide a single-cell nucleocytoplasmic profile in a high-throughput manner (similar to 100 cells per minute) for effective quantitative cell biology.
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