miR-122 direct detection in human serum by time-gated fluorescence imaging

Abstract

A new time-gated fluorescence-based imaging method to analyse microRNAs directly from human serum using dynamic chemistry labelling.

Simultaneous Detection of Drug-Induced Liver Injury Protein and microRNA Biomarkers Using Dynamic Chemical Labelling on a Luminex MAGPIX System

“…Prompted by these current analytical limitations, our group developed seqCOMBO, a new method to overcome the inability of current technology to analyse miRNAs without affecting proteins. In seqCOMBO, our DCL transformative technology to interrogate miRNAs [17][18][19][20][27][28][29][30][31][32][33] was combined with an antibody-dependant method on the Luminex MAGPIX system.…”

“…This first step, to hybridise the miR-122, was performed in a 96-well plate using a microplate orbital at 700 rpm for 1 h at 40 • C. After the hybridization, the DGL-122 beads were washed three times with the wash buffer. The DGL-122 beads were resuspended in 50 µL of assay buffer containing 5 µM SMART-C biotin and 1 mM sodium cyanoborohydride [17][18][19][20][27][28][29][30][31][32][33]. The 96-well plate was shaken at 700 rpm at 40 • C for 1 h. The beads were washed three more times and incubated with 70 µL of 2 µg/mL SA-PE for 5 min at 40 • C while being shaken at 700 rpm.…”

“…With the advances made by our team with dynamic chemical labelling (DCL) technology for the direct detection of nucleic acids, the deliverable of simultaneous detection of protein and miRNA in clinical samples is now possible. The DCL approach [17][18][19][20][27][28][29][30][31][32][33] is particularly well suited to deliver consistent and reliable quantitative readings of miRNAs in clinical samples when merged with bead-based systems. By simplifying the workflow, especially removing extraction, isolation and amplification steps, DCL was able to direct detect miRNAs in enzyme-linked immunosorbent assay (ELISA)-type format without affecting protein co-analytes, overcoming the current limitation issues that inhibit the development of simultaneous detection of proteins and miRNAs with high specificity and accuracy.…”

High resolution biosensor to test the capping level and integrity of mRNAs

“…( 18 ) proposed a biosensor able to analyse complete mRNAs, based on a bead-based silicon photonic resonator in conjunction with chaperone probes to reduce the mRNA secondary structure and enhance the access to the target. There are also biosensors focusing on the detection of specific miRNAs and mRNAs or even multiplexed detection of nucleic acids that have a great interest for diagnosis ( 19–21 ).…”

A fluorescent calix[4]arene with naphthalene units at the upper rim exhibits long fluorescence emission lifetime without fluorescence quenching

“…Also, only a few uorescent probes have achieved both high intensity and long lifetime. [13][14][15] In recent years, it was reported that bright and long-lived uorescence can be obtained from the excimer state 13,[16][17][18][19] that was conventionally thought to be the cause of quenching due to the low-energy excimer trap states with forbidden radiative transition and activated non-radiative process. [20][21][22][23][24] However, most of these reports were in the solid state, where the molecular movement is suppressed and the luminescence occurs in a single crystal.…”