A turn-on fluorescence sensor for rapid sensing of ATP based on luminescence resonance energy transfer between upconversion nanoparticles and Cy3 in vivo or vitro
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Keywords:
Biocompatibility
Nanosensor
Aptamer
Adenosine triphosphate
Photon Upconversion
Intracellular pH
Fluorescence-lifetime imaging microscopy
Colloidal quantum dots (QDs) possess numerous physical and optical properties that are ideal for biosensing and multiplexing applications. Fluorescence resonance energy transfer (FRET) is a well-established technique for detecting molecular-scale interactions due to proximity-driven changes in fluorescence. We have shown that QDs are excellent energy donors where dye labeled proteins serve as acceptors, and developed a number of prototype nanosensors for small molecules in solution. Among the more promising potential uses of QD-based FRET nanosensors is the ability to "multiplex" signal channels for parallel detection. Because of their very broad absorption and narrow symmetric emission spectra, QDs are ideal fluorophores for multiplexing applications. In this paper, we describe the benefits of QDs in FRET-based assays (as donors and acceptors) and the potential for signal multiplexing in nanoscale biosensors.
Nanosensor
Multiplex
Molecular biophysics
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Acid-sensing ion channels (ASICs) are trimeric cation-selective channels activated by decreases in extracellular pH. The intracellular N and C terminal tails of ASIC1 influence channel gating, trafficking, and signaling in ischemic cell death. Despite several X-ray and cryo-EM structures of the extracellular and transmembrane segments of ASIC1, these important intracellular tails remain unresolved. Here, we describe the coarse topography of the chicken ASIC1 intracellular domains determined by fluorescence resonance energy transfer (FRET), measured using either fluorescent lifetime imaging or patch clamp fluorometry. We find the C terminal tail projects into the cytosol by approximately 35 Å and that the N and C tails from the same subunits are closer than adjacent subunits. Using pH-insensitive fluorescent proteins, we fail to detect any relative movement between the N and C tails upon extracellular acidification but do observe axial motions of the membrane proximal segments toward the plasma membrane. Taken together, our study furnishes a coarse topographic map of the ASIC intracellular domains while providing directionality and context to intracellular conformational changes induced by extracellular acidification.
Acid-sensing ion channel
Intracellular pH
Cell membrane
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A novel DNA biosensor system on silica microspheres as solid carriers which based on the fluorescence resonance energy transfer (FRET) was presented in this work when CdTe quantum dots (QDs) were as energy donors and Au nanoparticles (AuNPs) were as energy accepters. Compared with the fluorescent intensity of CdTe QDs, the fluorescent intensity of DNA biosensors decreased extremely, which indicated that the FRET occurred between CdTe QDs and AuNPs. The biosensor system would have a certain degree recovery of fluorescence when the complementary single stranded DNA was introduced into this system. The DNA detection results indicated that this novel fluorescent DNA probe system could recognize the existence of complementary target DNA or not.
Cadmium telluride photovoltaics
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In this study, a turn-on nanosensor for detecting Hg2+ was developed based on the fluorescence resonance energy transfer (FRET) between long-strand aptamers-functionalized upconversion nanoparticles (UCNPs) and short-strand aptamers-functionalized gold nanoparticles (GNPs). In the absence of Hg2+, FRET between UCNPs and GNPs occurred because of the specific matching between two aptamers, resulting in the fluorescence quenching of UCNPs. In the presence of Hg2+, long-stranded aptamers fold back into a hairpin structure due to the stable binding interactions between Hg2+ and thymine, leading to the release of GNPs from UCNPs, resulting in the quenched fluorescence restoration. Under the optimized conditions, the nanosensor achieved a linear detection range of 0.2–20 μM and a low detection limit (LOD) of 60 nM. Meanwhile, it showed good selectivity and has been applied to detecting Hg2+ in tap water and milk samples with good precision.
Aptamer
Nanosensor
Photon Upconversion
Linear range
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During the last two decades, porous silicon (PSi) has been proposed as a high-performance biosensing platform due to its biocompatibility, surface tailorability and reproducibility. This review focuses on the recent developments and progress in the area related to hybrid PSi biosensors using plasmonic metal nanoparticles (MNPs), fluorescent quantum dots (QDs) or a combination of both MNPs and QDs for creating hybrid nanostructured architectures for ultrasensitive detection of biomolecules. The review discusses the mechanisms of sensitivity enhancement based on Localized Surface Plasmon Resonance (LSPR) of MNPs, Fluorescence Resonance Energy Transfer (FRET) in the case of MNPs/QDs donor-acceptor interactions, and photoluminescence/ fluorescence enhancement resulting from the embedded fluorescent QDs inside the PSi microcavity. The review highlights the key features of hybrid PSi/ MNPs/ QDs biosensors for dual-mode detection applications.
Nanosensor
Nanomaterials
Biomolecule
Biocompatibility
Porous Silicon
Surface Modification
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The formation of extracellular amyloid-β (Aβ) plaques is a common molecular change that underlies several debilitating human conditions, including Alzheimer's disease (AD); however, the existing near-infrared (NIR) fluorescent probes for the in vivo detection of Aβ plaques are limited by undesirable fluorescent properties and poor brain kinetics. In this work, we designed, synthesized, and evaluated a new family of efficient NIR probes that target Aβ plaques by incorporating hydroxyethyl groups into the ligand structure. Among these probes, DANIR 8c showed excellent fluorescent properties with an emission maximum above 670 nm upon binding to Aβ aggregates and also displayed a high sensitivity (a 629-fold increase in fluorescence intensity) and affinity (Kd = 14.5 nM). Because of the improved hydrophilicity that was induced by hydroxyls, 8c displayed increased initial brain uptake and a fast washout from the brain, as well as an acceptable biostability in the brain. In vivo NIR fluorescent imaging revealed that 8c could efficiently distinguish between AD transgenic model mice and normal controls. Overall, 8c is an efficient and veritable NIR fluorescent probe for the in vivo detection of Aβ plaques in the brain.
Amyloid (mycology)
Fluorescence-lifetime imaging microscopy
Autofluorescence
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Aptamer
Fluorescence-lifetime imaging microscopy
Internalization
Adenosine triphosphate
Live cell imaging
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Owing to the significant roles of adenosine triphosphate (ATP) in diverse biological processes, ATP level is used to research and evaluate the physiological processes of organisms. Aptamer-based biosensors have been widely reported to achieve this purpose, which are superior in their flexible biosensing mechanism, with a high sensitivity and good biocompatibility; however, the aptamers currently used for ATP detection have a poor ability to discriminate ATP from adenosine diphosphate (ADP) and adenosine monophosphate (AMP). Herein, an ATP-specific aptamer was screened and applied to construct a fluorescent aptasensor for ATP by using graphene oxide (GO) and strand displacement amplification (SDA). The fluorescence intensity of the sensor is linearly related to the concentration of ATP within 0.1 μM to 25 μM under optimal experimental conditions, and the detection limit is 33.85 nM. The biosensor exhibits a satisfactory specificity for ATP. Moreover, the experimental results indicate that the biosensor can be applied to determine the ATP in human serum. In conclusion, the screened aptamer and the biosensor have promising applications in the determination of the real energy charge level and ATP content in a complex biological system.
Aptamer
Adenosine triphosphate
Adenosine monophosphate
Adenosine diphosphate
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A versatile pH-dependent fluorescent protein was applied to intracellular pH measurements by means of the phasor approach to fluorescence lifetime imaging. By this fit-less method we obtain intracellular pH maps under resting or altered physiological conditions by single-photon confocal or two-photon microscopy.
Fluorescence-lifetime imaging microscopy
Intracellular pH
Two-photon excitation microscopy
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Carbon dots were synthesized by a simple and green strategy for selective and sensitive Cu(2+) ion detection using both down and upconversion fluorescence. These fluorescent nanosensors show low cytotoxicity and are applied for intracellular sensing and imaging of Cu(2+) in biological systems.
Nanosensor
Photon Upconversion
Carbon fibers
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Citations (269)