Fluorescent nanodiamond (FND) has recently played a central role in fueling new discoveries in interdisciplinary fields spanning biology, chemistry, physics, and materials sciences. The nanoparticle is unique in that it contains a high density ensemble of negatively charged nitrogen-vacancy (NV(-)) centers as built-in fluorophores. The center possesses a number of outstanding optical and magnetic properties. First, NV(-) has an absorption maximum at ∼550 nm, and when exposed to green-orange light, it emits bright fluorescence at ∼700 nm with a lifetime of longer than 10 ns. These spectroscopic properties are little affected by surface modification but are distinctly different from those of cell autofluorescence and thus enable background-free imaging of FNDs in tissue sections. Such characteristics together with its excellent biocompatibility render FND ideal for long-term cell tracking applications, particularly in stem cell research. Next, as an artificial atom in the solid state, the NV(-) center is perfectly photostable, without photobleaching and blinking. Therefore, the NV-containing FND is suitable as a contrast agent for super-resolution imaging by stimulated emission depletion (STED). An improvement of the spatial resolution by 20-fold is readily achievable by using a high-power STED laser to deplete the NV(-) fluorescence. Such improvement is crucial in revealing the detailed structures of biological complexes and assemblies, including cellular organelles and subcellular compartments. Further enhancement of the resolution for live cell imaging is possible by manipulating the charge states of the NV centers. As the "brightest" member of the nanocarbon family, FND holds great promise and potential for bioimaging with unprecedented resolution and precision. Lastly, the NV(-) center in diamond is an atom-like quantum system with a total electron spin of 1. The ground states of the spins show a crystal field splitting of 2.87 GHz, separating the ms = 0 and ±1 sublevels. Interestingly, the transitions between the spin sublevels can be optically detected and manipulated by microwave radiation, a technique known as optically detected magnetic resonance (ODMR). In addition, the electron spins have an exceptionally long coherence time, making FND useful for ultrasensitive detection of temperature at the nanoscale. Pump-probe-type nanothermometry with a temporal resolution of better than 10 μs has been achieved with a three-point sampling method. Gold/diamond nanohybrids have also been developed for highly localized hyperthermia applications. This Account provides a summary of the recent advances in FND-enabled technologies with a special focus on long-term cell tracking, super-resolution imaging, and nanoscale temperature sensing. These emerging and multifaceted technologies are in synchronicity with modern imaging modalities.
Measuring temperature in nanoscale spatial resolution either at or far from equilibrium is of importance in many scientific and technological applications. Although negatively charged nitrogen-vacancy (NV(-)) centers in diamond have recently emerged as a promising nanometric temperature sensor, the technique has been applied only under steady state conditions so far. Here, we present a three-point sampling method that allows real-time monitoring of the temperature changes over ±100 K and a pump-probe-type experiment that enables the study of nanoscale heat transfer with a temporal resolution of better than 10 μs. The utility of the time-resolved luminescence nanothermometry was demonstrated with 100 nm fluorescent nanodiamonds spin-coated on a glass substrate and submerged in gold nanorod solution heated by a near-infrared laser, and the validity of the measurements was verified with finite-element numerical simulations. The combined theoretical and experimental approaches will be useful to implement time-resolved temperature sensing in laser processing of materials and even for devices in operation at the nanometer scale.
Much of the current understanding of thermal effects in biological systems is based on macroscopic measurements. There is little knowledge about the local thermostability or heat tolerance of subcellular components at the nanoscale. Herein, we show that gold nanorod-fluorescent nanodiamond (GNR-FND) hybrids are useful as a combined nanoheater/nanothermometer in living cells. With the use of a 594 nm laser for both heating and probing, we measure the temperature changes by recording the spectral shifts of the zero-phonon lines of negatively charged nitrogen-vacancy centers in FNDs. The technique allows us to determine the rupture temperatures of individual membrane nanotubes in human embryonic kidney cells, as well as to generate high temperature gradients on the cell membrane for photoporation and optically controlled hyperthermia. Our results demonstrate a new paradigm for hyperthermia research and application.
Highly stable lipid-encapsulated fluorescent nanodiamonds (FNDs) are produced by photo-crosslinking of diacetylene-containing lipids physically attached to the FND surface. Not only is this coating method simple and fast, but also it gives the FND-lipid hybrids favorable properties for bioapplications. The hybrids are useful as fluorescent biolabels as well as fiducial markers for correlative light and electron microscopy.
Recent advances in quantum technology have demonstrated the potential use of negatively charged nitrogen-vacancy (NV -) centers in diamond for temperature and magnetic sensing at sub-cellular levels. Fluorescent nanodiamonds (FNDs) containing high-density ensembles of NV -centers are appealing for such applications because they are inherently biocompatible and non-toxic. Here, we show that FNDs conjugated with gold nanorods (GNRs) are useful as a combined nanoheater and nanothermometer for highly localized hyperthermia treatment using near-infrared (NIR) lasers as the heating source. A temperature rise of ∼10 K can be readily achieved at a NIR laser power of 0.4 mW in cells. The technique is compatible with the presence of static magnetic fields and allows for simultaneous temperature and magnetic sensing with nanometric spatial resolution. To elucidate the nanoscale heating process, numerical simulations are conducted with finite element analysis, providing an important guideline for the use of this new tool for active and high-precision control of temperature under diverse environmental conditions.
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