期刊
JOURNAL OF NANOBIOTECHNOLOGY
卷 20, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s12951-022-01274-3
关键词
Ratiometric fluorescence; Microfluidic; MicroRNA-21; Dual emissive assay; Carbon dots; Cell imaging
资金
- research office of the University of Kurdistan [98.11.2533]
- Iranian Nanotechnology Initiative
A strategy for constructing a dual-emission fluorescent sensor using carbon dots was reported for ratiometric microRNA-21 sensing and bioimaging of cancer cells. The sensor showed a wide linear range and low detection limit, demonstrating potential for clinical diagnosis. Additionally, the sensor successfully detected microRNA-21 in diluted human serum samples and MCF-7 cell lines, showcasing its versatility in potential clinical applications.
Background: MicroRNAs have short sequences of 20 similar to 25-nucleotides which are similar among family members and play crucial regulatory roles in numerous biological processes, such as in cell development, metabolism, proliferation, differentiation, and apoptosis. Results: We reported a strategy for the construction of a dual-emission fluorescent sensor using carbon dots (CDs) and confirmed their applications for ratiometric microRNA-21 sensing and bioimaging of cancer cells in a microfluidic device. The composition of blue CDs (B-CDs) and yellow CDs (Y-CDs) depicts dual-emission behavior which is centered at 409 and 543 nm under an excitation wavelength of 360 nm. With increasing microRNA-21 concentration, the robust and specific binding of DNA probe functionalized B-CDs to complementary microRNA-21 target induced perturbations of probe structure and led to changing fluorescence intensity in both wavelengths. Consequently, the ratio of turn-on signal to turn-off signal is greatly altered. With monitoring of the inherent ratiometric fluorescence variation (Delta F-540nm/Delta F-410nm), as-prepared BY-CDs were established as an efficient platform for ratiometric fluorescent microRNA-21 sensing, with a wide linear range of 0.15 fM to 2.46 pM and a detection limit of 50 aM. Conclusions: Furthermore, the proposed assay was applied for detecting microRNA-21 in dilute human serum samples with satisfactory recovery and also in MCF-7 cell lines in the range 3000 to 45,000 (cell mL(-1)) with a detection limit (3 cells in 10 mu L), demonstrating the potential of the assay for clinic diagnosis of microRNA-associated disease. More importantly, the images revealed that MCF-7 cells well labeled with BY-CDs could exhibit the applicability of the proposed microfluidic system as an effective cell trapping device in bioimaging.
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