BUSCADOR RECOLECTA

MicroRNAs (miRs) have emerged as promising biomarkers for early disease diagnosis and personalised treatment monitoring. However, but their clinical utility has been hampered by technical limitations. Dynamic chemical labelling (DCL) based on capturing abasic PNA probes and reactive nucleobases, so-called SMART bases, is a PCR-free approach that has proven very useful for the direct interrogation of circulating miRs. In this work, we expand the palette of available tools for DCL miR detection methods with a newly synthesised SMART nucleobase, SMART-C-Eu. This nucleobase contains a stable lanthanide cryptate. Using this SMART-C-Eu base and time-gated (TG) luminescence imaging, we successfully detect and quantify miR-122-5p in human serum samples. miR-122-5p is a well-known biomarker for drug-induced liver injury. A bead-counting analysis approach improved statistical robustness and allowed the detection of miR-122-5p concentrations in the nanomolar range. Furthermore, we extend this approach to multiplexed detection of three different miRs (miR-371a-3p, miR-451a-5p, and miR-122-5p) using spectral and temporal filtering. Consistent results were obtained using machine learning algorithms for automatic bead classification. Although this technique requires further sensitivity improvements to detect miRs at lower concentrations, it represents a significant advancement in miR analysis. It offers multiplexing capabilities and the potential for automation, paving the way for more accurate and robust clinical applications in the future., European Union through the H2020 diaRNAgnosis project, under grant agreement No. 101007934, Agencia Estatal de Investigación (Spain) through grant PID2020-114256RB-I00 AEI/10.13039/501100011033, Agencia Estatal de Investigación (Spain) through grant CTQ2017-85658-R AEI/10.13039/501100011033/FEDER “Una manera de hacer Europa”.

This work was supported by grant CTQ2017-85658-R funded by MCIN/AEI/10.13039/501100011033/ FEDER “Una manera de hacer Europa”, grant PID2020-114256RB-I00 funded by MCIN/AEI/10.13039/501100011033, and the diaRNAgnosis project funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 101007934. P.M.R.P. acknowledges funding from Fundação para a Ciência e a Tecnologia, I.P., through projects UIDB/00100/2020 and 2022.04076.PTDC., Controlling the nano- and micropatterning of metal structures is an important requirement for various technological applications in photonics and biosensing. We present a method for controllably creating silver micropatterns by laser-induced photo-sculpting. Photo-sculpting is driven by plasmonic interactions between pulsed laser radiation and silver nanorods (AgNRs) in aqueous suspension; this process leads to optical binding forces transporting the AgNRs in the surroundings, while electronic thermalization results in photooxidation, melting, and ripening of the AgNRs into well-defined 3D structures. We called these structures Airy castles due to their structural similarity with a diffraction-limited Airy disk. The photo-sculpted Airy castles contain emissive Ag nanoclusters, allowing for the visualization and examination of the aggregation process using luminescence microscopy. We comprehensively examine the factors that define the photo-sculpting process, namely, the concentration and shape of the AgNRs, as well as the energy, power and repetition rate of the laser. Finally, we investigate the potential applications by measuring the metal-enhanced luminescence of a europium-based luminophore using Airy castles., Grant CTQ2017-85658-R funded by MCIN/AEI/10.13039/501100011033/ FEDER “Una manera de hacer Europa”, Grant PID2020-114256RB-I00 funded by MCIN/AEI/10.13039/501100011033, diaRNAgnosis project funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 101007934, Fundação para a Ciência e a Tecnologia, I.P., through projects UIDB/00100/2020 and 2022.04076.PTDC