Ratiometric fluorescent sensing of pyrophosphate with sp³-functionalized single-walled carbon nanotubes.

Inorganic pyrophosphate is a key molecule in many biological processes from DNA synthesis to cell metabolism. Here we introduce sp3-functionalized (6,5) single-walled carbon nanotubes (SWNTs) with red-shifted defect emission as near-infrared luminescent probes for the optical detection and quantification of inorganic pyrophosphate. The sensing scheme is based on the immobilization of Cu2+ ions on the SWNT surface promoted by coordination to covalently attached aryl alkyne groups and a triazole complex. The presence of Cu2+ ions on the SWNT surface causes fluorescence quenching via photoinduced electron transfer, which is reversed by copper-complexing analytes such as pyrophosphate. The differences in the fluorescence response of sp3-defect to pristine nanotube emission enables reproducible ratiometric measurements in a wide concentration window. Biocompatible, phospholipid-polyethylene glycol-coated SWNTs with such sp3 defects are employed for the detection of pyrophosphate in cell lysate and for monitoring the progress of DNA synthesis in a polymerase chain reaction. This robust ratiometric and near-infrared luminescent probe for pyrophosphate may serve as a starting point for the rational design of nanotube-based biosensors.

Stochastic Formation of Quantum Defects in Carbon Nanotubes.

Small perturbations in the structure of materials significantly affect their properties. One example is single wall carbon nanotubes (SWCNTs), which exhibit chirality-dependent near-infrared (NIR) fluorescence. They can be modified with quantum defects through the reaction with diazonium salts, and the number or distribution of these defects determines their photophysics. However, the presence of multiple chiralities in typical SWCNT samples complicates the identification of defect-related emission features. Here, we show that quantum defects do not affect aqueous two-phase extraction (ATPE) of different SWCNT chiralities into different phases, which suggests low numbers of defects. For bulk samples, the bandgap emission (E11) of monochiral (6,5)-SWCNTs decreases, and the defect-related emission feature (E11*) increases with diazonium salt concentration and represents a proxy for the defect number. The high purity of monochiral samples from ATPE allows us to image NIR fluorescence contributions (E11 = 986 nm and E11* = 1140 nm) on the single SWCNT level. Interestingly, we observe a stochastic (Poisson) distribution of quantum defects. SWCNTs have most likely one to three defects (for low to high (bulk) quantum defect densities). Additionally, we verify this number by following single reaction events that appear as discrete steps in the temporal fluorescence traces. We thereby count single reactions via NIR imaging and demonstrate that stochasticity plays a crucial role in the optical properties of SWCNTs. These results show that there can be a large discrepancy between ensemble and single particle experiments/properties of nanomaterials.

Guanine Quantum Defects in Carbon Nanotubes for Biosensing.

Fluorescent single-wall carbon nanotubes (SWCNTs) are used as nanoscale biosensors in diverse applications. Selectivity is built in by noncovalent functionalization with polymers such as DNA. Recently, covalent functionalization was demonstrated by conjugating guanine bases of adsorbed DNA to the SWCNT surface as guanine quantum defects (g-defects). Here, we create g-defects in (GT)10-coated SWCNTs (Gd-SWCNTs) and explore how this affects molecular sensing. We vary the defect densities, which shifts the E11 fluorescence emission by 55 nm to a λmax of 1049 nm. Furthermore, the Stokes shift between absorption and emission maximum linearly increases with defect density by up to 27 nm. Gd-SWCNTs represent sensitive sensors and increase their fluorescence by >70% in response to the important neurotransmitter dopamine and decrease it by 93% in response to riboflavin. Additionally, the extent of cellular uptake of Gd-SWCNTs decreases. These results show how physiochemical properties change with g-defects and that Gd-SWCNTs constitute a versatile optical biosensor platform.