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1.
Angew Chem Int Ed Engl ; 63(9): e202316965, 2024 Feb 26.
Article in English | MEDLINE | ID: mdl-38100133

ABSTRACT

Enzymatic reactions are used to detect analytes in a range of biochemical methods. To measure the presence of an analyte, the enzyme is conjugated to a recognition unit and converts a substrate into a (colored) product that is detectable by visible (VIS) light. Thus, the lowest enzymatic turnover that can be detected sets a limit on sensitivity. Here, we report that substrates and products of horseradish peroxidase (HRP) and ß-galactosidase change the near-infrared (NIR) fluorescence of (bio)polymer modified single-walled carbon nanotubes (SWCNTs). They translate a VIS signal into a beneficial NIR signal. Moreover, the affinity of the nanosensors leads to a higher effective local concentration of the reactants. This causes a non-linear sensor-based signal amplification and translation (SENSAT). We find signal enhancement up to ≈120x for the HRP substrate p-phenylenediamine (PPD), which means that reactions below the limit of detection in the VIS can be followed in the NIR (≈1000 nm). The approach is also applicable to other substrates such as 3,3'-5,5'-tetramethylbenzidine (TMB). An adsorption-based theoretical model fits the observed signals and corroborates the sensor-based enhancement mechanism. This approach can be used to amplify signals, translate them into the NIR and increase sensitivity of biochemical assays.


Subject(s)
Nanotubes, Carbon , Light , Horseradish Peroxidase , Adsorption , Biological Assay
2.
ACS Appl Bio Mater ; 6(1): 309-317, 2023 01 16.
Article in English | MEDLINE | ID: mdl-36538701

ABSTRACT

Fluorophores emitting in the near-infrared (NIR) wavelength region present optimal characteristics for photonics and especially bioimaging. Unfortunately, only few NIR fluorescent materials are known, and even fewer are biocompatible. For this reason, the scientific interest in designing NIR fluorophores is very high. Egyptian Blue (CaCuSi4O10, EB) is an NIR fluorescent layered silicate that can be exfoliated into fluorescent nanosheets (EB-NS). So far, its surface chemistry has not been tailored, but this is crucial for colloidal stability and biological targeting. Here, we demonstrate covalent surface functionalization of EB nanosheets (EBfunc) via Si-H activation using hydrosilanes with variable functionalities. In the first part of this work, EB-NS are grafted with the visible fluorescent pyrene (Pyr) moieties to demonstrate conjugation by colocalization of the Vis/NIR fluorescence on the (single) EB-NS level. Next, the same grafting procedure was repeated and validated with carboxyl group (COOH)-containing hydrosilanes. These groups serve as a generic handle for further (bio)functionalization of the EB-NS surface. In this way, folic acid (FA) could be conjugated to EB-NS, allowing the targeting of folic acid receptor-expressing cancer cells. These results highlight the potential of this surface chemistry approach to modify EB-NS, enabling targeted NIR imaging for biomedical applications.


Subject(s)
Fluorescent Dyes , Silicates , Copper , Folic Acid
3.
Proc Natl Acad Sci U S A ; 119(22): e2202842119, 2022 05 31.
Article in English | MEDLINE | ID: mdl-35613050

ABSTRACT

The neurotransmitter dopamine (DA) controls multiple behaviors and is perturbed in several major brain diseases. DA is released from large populations of specialized structures called axon varicosities. Determining the DA release mechanisms at such varicosities is essential for a detailed understanding of DA biology and pathobiology but has been limited by the low spatial resolution of DA detection methods. We used a near-infrared fluorescent DA nanosensor paint, adsorbed nanosensors detecting release of dopamine (AndromeDA), to detect DA secretion from cultured murine dopaminergic neurons with high spatial and temporal resolution. We found that AndromeDA detects discrete DA release events and extracellular DA diffusion and observed that DA release varies across varicosities. To systematically detect DA release hotspots, we developed a machine learning­based analysis tool. AndromeDA permitted the simultaneous visualization of DA release for up to 100 dopaminergic varicosities, showing that DA release hotspots are heterogeneous and occur at only ∼17% of all varicosities, indicating that many varicosities are functionally silent. Using AndromeDA, we determined that DA release requires Munc13-type vesicle priming proteins, validating the utility of AndromeDA as a tool to study the molecular and cellular mechanism of DA secretion.


Subject(s)
Axons , Dopamine , Dopaminergic Neurons , Nanostructures , Neurotransmitter Agents , Optical Imaging , Animals , Axons/metabolism , Brain/metabolism , Dopamine/analysis , Dopamine/metabolism , Dopaminergic Neurons/metabolism , Fluorescent Dyes/chemistry , Mice , Neurotransmitter Agents/analysis , Neurotransmitter Agents/metabolism , Optical Imaging/methods , Paint , Spectroscopy, Near-Infrared/methods
4.
Acta Crystallogr Sect E Struct Rep Online ; 66(Pt 2): i13, 2010 Jan 30.
Article in English | MEDLINE | ID: mdl-21579587

ABSTRACT

In contrast to the previous structure determinations of the title structure, (NH(4))(2)[MoS(4)], the present determination at 173 K localized the positions of the H atoms. The title structure belongs to the ß-K(2)SO(4) family and all the ions are located on crystallographic mirror planes. The ions are held together by N-H⋯S hydrogen bonds (some of which are bifurcated), forming a three-dimensional network. One of the N atoms has nine contacts to the S atoms shorter than 4 Å, and the other has ten.

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