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.
Diamond has received increasing attention for its promising biomedical applications. The material is highly biocompatible and can be easily conjugated with bioactive molecules. Recently, nanoscale diamond has been applied as light scattering labels and luminescent optical markers. The luminescence, arising from photoexcitation of colour centres, can be substantially enhanced when type Ib diamond nanocrystals are bombarded by a high-energy particle beam and then annealed to form negatively charged nitrogen-vacancy centres. The centre absorbs strongly at 560 nm, fluoresces efficiently in the far-red region and is exceptionally photostable (without photoblinking and photobleaching). It is an ideal candidate for long-term imaging and tracking in complex cellular environments. This review summarizes recent advances in the development of fluorescent nanodiamonds for optical bioimaging with single particle sensitivity and nanometric resolution.
Cell therapy is a promising strategy for the treatment of human diseases. While the first use of cells for therapeutic purposes can be traced to the 19th century, there has been a lack of general and reliable methods to study the biodistribution and associated pharmacokinetics of transplanted cells in various animal models for preclinical evaluation. Here, we present a new platform using albumin-conjugated fluorescent nanodiamonds (FNDs) as biocompatible and photostable labels for quantitative tracking of human placenta choriodecidual membrane-derived mesenchymal stem cells (pcMSCs) in miniature pigs by magnetic modulation. With this background-free detection technique and time-gated fluorescence imaging, we have been able to precisely determine the numbers as well as positions of the transplanted FND-labeled pcMSCs in organs and tissues of the miniature pigs after intravenous administration. The method is applicable to single-cell imaging and quantitative tracking of human stem/progenitor cells in rodents and other animal models as well.
Dynamics of fluorescent diamond nanoparticles in HeLa cells has been studied with two-photon fluorescence correlation spectroscopy (FCS). Fluorescent nanodiamond (FND) is an excellent fluorescent probe for bioimaging application, but they are often trapped in endosomes after cellular uptake. The entrapment prohibits FCS from being performed in a time frame of 60 s. Herein, we show that the encapsulation of FNDs within a lipid layer enhances the diffusion of the particles in the cytoplasm by more than one order of magnitude, and particles as small as 40 nm can be probed individually with high image contrast by two-photon excited luminescence. The development of the technique together with single particle tracking through one-photon excitation allows probing of both short-term and long-term dynamics of single FNDs in living cells.
Fluorescent nanodiamonds (FNDs) of 10–20 nm in size have been produced by helium ion irradiation and isolated by differential centrifugation. Spectroscopic characterization of the FNDs with fluorescence microscopy and fluorescence correlation spectroscopy indicates perfect photostability and use as an alternative to far‐red fluorescent proteins for biolabeling and fluorescence resonance energy transfer applications.
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