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In this study, an unreported Ir(III) complex 1 was identified by screening as a versatile G-quadruplex probe. It exhibited highly selective response for different G-quadruplex DNA over double strand, single strand and triplex DNA. Compared with the organic G-quadruplex probe thioflavin T, complex 1 displays a longer lifetime, a larger Stokes shift, comparable G-quadruplex/ssDNA enhancement ratios, and higher G-quadruplex/triplex DNA enhancement ratios. In consideration of the encouraging G-quadruplex probe performance of complex 1, we employed 1 to develop a G-quadruplex-based detection system for the detection of insulin as a "proof-of-principle" concept. We also demonstrate an optimization process that enhanced the sensitivity of this sensing assay. Compared to previously reported methods, our "mix-and-detect" detection methodology is easy operated, quick, and cost-effective. A detection limit as low as 80 pM for insulin can be achieved by this sensing approach, with a linear relationship between luminescence intensity and insulin concentration established from 80 pM to 20 nM. Moreover, this assay could work effectively in diluted human serum.
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http://dx.doi.org/10.1021/acs.analchem.5b04064 | DOI Listing |
Asian J Androl
September 2025
Joint Laboratory of Reproductive Medicine, Sichuan University-Chinese University of Hong Kong (SCU-CUHK), Key Laboratory of Obstetric, Gynaecologic and Paediatric Diseases and Birth Defects of Ministry of Education, West China Second University Hospital, Med-X Centre for Manufacturing, Sichuan Unive
In the evaluation of male infertility, precise assessment of sperm functional competence has surpassed the requirements of conventional semen parameters. Existing computer-aided analysis systems are deficient at the molecular diagnostic level and also face challenges in live-cell fluorescence quantification. To address these issues, we have developed a novel integrated computational-imaging platform that combines a fine-tuned You Only Look Once version 8 (YOLOv8) architecture, tailored for the EVISEN dataset, with dual-probe fluorescence microscopy image segmentation, enabling simultaneous quantification of intracellular pH (pHi) and mitochondrial DNA G-quadruplexes (mtDNA G4s).
View Article and Find Full Text PDFAnal Chem
September 2025
Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
Label-free fluorescent binding assays employing DNA staining dyes as probes are widely adopted techniques in the aptamer field. While many dyes have been used, thioflavin T (ThT) did not receive much attention for this purpose until recently, since it has long been perceived primarily as a G-quadruplex staining dye. Based on recent studies, ThT appears to serve as a reliable probe for evaluating the binding of non-G-quadruplex aptamers, and we seek to clarify the underlying mechanisms responsible for the exceptional performance of ThT.
View Article and Find Full Text PDFTalanta
August 2025
School of Pharmacy, Jiangsu University, Zhenjiang, 212013, PR China.
G-Quadruplex (G4) DNA structures play a crucial role in regulating various biological processes, rendering them attractive targets for diagnostic and therapeutic applications. The development of G4-targeted fluorescent probes will significantly enhance our understanding of G4 DNA biology in vivo and improve the precision of diagnosing and treating genetic diseases. Herein, we report on a regioisomer of a thiazole orange derivative (SQ2) as a promising fluorescent dye for G4 DNA structures and demonstrate its application in both cell and tumor tissue imaging.
View Article and Find Full Text PDFBiosens Bioelectron
December 2025
University of Science and Technology of China, Hefei, 230026, China; Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, 215163, China; Tianjin Guoke Medical Technology Development Co., Ltd, Tianjin, 300399, China. Electronic address:
In conventional electrochemical biosensors, nanobubbles generated by electrochemical reactions typically adhere to the electrode interface, which affect the accuracy of measurements. In this study, we take good use of nanobubbles as the signal source and effectively address this limitation. An ultrasensitive approach for the detection of miRNA is further developed coupling catalytic hairpin assembly and DNA-functionalized nanopores to generate nanobubbles.
View Article and Find Full Text PDFAnal Chem
September 2025
Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, Zhejiang 310022, China.
Cell-membrane-anchored DNA probes are powerful tools for membrane-localized biosensing, bioregulation, and biotherapy studies. However, the poor biostability, membrane-anchoring stability, and effectiveness of membrane-anchored DNA probes in complex physiological environments have seriously hindered their widespread applications. In this work, probe-encapsulated DNA nanotoolboxes were developed by immobilizing DNA probes into DNA cuboids to improve the performance of cell-membrane-anchored DNA probes in complex biological media.
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