Targeted nanodiamonds for identification of subcellular protein assemblies in mammalian cells.

Transmission electron microscopy (TEM) can be used to successfully determine the structures of proteins. However, such studies are typically done ex situ after extraction of the protein from the cellular environment. Here we describe an application for nanodiamonds as targeted intensity contrast labels in biological TEM, using the nuclear pore complex (NPC) as a model macroassembly. We demonstrate that delivery of antibody-conjugated nanodiamonds to live mammalian cells using maltotriose-conjugated polypropylenimine dendrimers results in efficient localization of nanodiamonds to the intended cellular target. We further identify signatures of nanodiamonds under TEM that allow for unambiguous identification of individual nanodiamonds from a resin-embedded, OsO4-stained environment. This is the first demonstration of nanodiamonds as labels for nanoscale TEM-based identification of subcellular protein assemblies. These results, combined with the unique fluorescence properties and biocompatibility of nanodiamonds, represent an important step toward the use of nanodiamonds as markers for correlated optical/electron bioimaging.

Nanodiamond landmarks for subcellular multimodal optical and electron imaging.

There is a growing need for biolabels that can be used in both optical and electron microscopies, are non-cytotoxic, and do not photobleach. Such biolabels could enable targeted nanoscale imaging of sub-cellular structures, and help to establish correlations between conjugation-delivered biomolecules and function. Here we demonstrate a sub-cellular multi-modal imaging methodology that enables localization of inert particulate probes, consisting of nanodiamonds having fluorescent nitrogen-vacancy centers. These are functionalized to target specific structures, and are observable by both optical and electron microscopies. Nanodiamonds targeted to the nuclear pore complex are rapidly localized in electron-microscopy diffraction mode to enable "zooming-in" to regions of interest for detailed structural investigations. Optical microscopies reveal nanodiamonds for in-vitro tracking or uptake-confirmation. The approach is general, works down to the single nanodiamond level, and can leverage the unique capabilities of nanodiamonds, such as biocompatibility, sensitive magnetometry, and gene and drug delivery.

NMR probe of metallic states in nanoscale topological insulators.

A 125Te NMR study of bismuth telluride nanoparticles as a function of particle size revealed that the spin-lattice relaxation is enhanced below 33 nm, accompanied by a transition of NMR spectra from the single to the bimodal regime. The satellite peak features a negative Knight shift and higher relaxivity, consistent with core polarization from p-band carriers. Whereas nanocrystals follow a Korringa law in the range 140-420 K, micrometer particles do so only below 200 K. The results reveal increased metallicity of these nanoscale topological insulators in the limit of higher surface-to-volume ratios.

Real-time maps of fluid flow fields in porous biomaterials.

Mechanical forces such as fluid shear have been shown to enhance cell growth and differentiation, but knowledge of their mechanistic effect on cells is limited because the local flow patterns and associated metrics are not precisely known. Here we present real-time, non-invasive measures of local hydrodynamics in 3D biomaterials based on nuclear magnetic resonance. Microflow maps were further used to derive pressure, shear and fluid permeability fields. Finally, remodeling of collagen gels in response to precise fluid flow parameters was correlated with structural changes. It is anticipated that accurate flow maps within 3D matrices will be a critical step towards understanding cell behavior in response to controlled flow dynamics.