Detecting Biothreat Agents: From Current Diagnostics to Developing Sensor Technologies.

Although a fundamental understanding of the pathogenicity of most biothreat agents has been elucidated and available treatments have increased substantially over the past decades, they still represent a significant public health threat in this age of (bio)terrorism, indiscriminate warfare, pollution, climate change, unchecked population growth, and globalization. The key step to almost all prevention, protection, prophylaxis, post-exposure treatment, and mitigation of any bioagent is early detection. Here, we review available methods for detecting bioagents including pathogenic bacteria and viruses along with their toxins. An introduction placing this subject in the historical context of previous naturally occurring outbreaks and efforts to weaponize selected agents is first provided along with definitions and relevant considerations. An overview of the detection technologies that find use in this endeavor along with how they provide data or transduce signal within a sensing configuration follows. Current "gold" standards for biothreat detection/diagnostics along with a listing of relevant FDA approved in vitro diagnostic devices is then discussed to provide an overview of the current state of the art. Given the 2014 outbreak of Ebola virus in Western Africa and the recent 2016 spread of Zika virus in the Americas, discussion of what constitutes a public health emergency and how new in vitro diagnostic devices are authorized for emergency use in the U.S. are also included. The majority of the Review is then subdivided around the sensing of bacterial, viral, and toxin biothreats with each including an overview of the major agents in that class, a detailed cross-section of different sensing methods in development based on assay format or analytical technique, and some discussion of related microfluidic lab-on-a-chip/point-of-care devices. Finally, an outlook is given on how this field will develop from the perspective of the biosensing technology itself and the new emerging threats they may face.

Unconventional slowing down of electronic recovery in photoexcited charge-ordered La1/3Sr2/3FeO3.

The coupling of ordered electronic phases with lattice, spin, and orbital degrees of freedom are of central interest in strongly correlated systems. Their interplay has been intensively studied from femtosecond to picosecond time scales, while their dynamics beyond nanoseconds are usually assumed to follow lattice cooling. Here, we report an unusual slowing down of the recovery of an electronic phase across a first-order phase transition. Following optical excitation, the recovery time of both transient optical reflectivity and X-ray diffraction intensity from the charge-ordered superstructure in a La1/3Sr2/3FeO3 thin film increases by orders of magnitude as the sample temperature approaches the phase transition temperature. In this regime, the recovery time becomes much longer than the lattice cooling time. The combined experimental and theoretical investigation shows that the slowing down of electronic recovery corresponds to the pseudo-critical dynamics that originates from magnetic interactions close to a weakly first-order phase transition.

Elevated Temperature Photophysical Properties and Morphological Stability of CdSe and CdSe/CdS Nanoplatelets.

Elevated temperature optoelectronic performance of semiconductor nanomaterials remains an important issue for applications. Here we examine 2D CdSe nanoplatelets (NPs) and CdS/CdSe/CdS shell/core/shell sandwich NPs at temperatures ranging from 300 to 700 K using static and transient spectroscopies as well as in situ transmission electron microscopy. NPs exhibit reversible changes in PL intensity, spectral position, and emission line width with temperature elevation up to ∼500 K, losing a factor of ∼8 to 10 in PL intensity at 400 K relative to ambient. Temperature elevation above ∼500 K yields thickness-dependent, irreversible degradation in optical properties. Electron microscopy relates stability of the core-only NP morphology up to 555 and 600 K for the four and five monolayer NPs, respectively, followed by sintering and evaporation at still higher temperatures. Reversible PL loss, based on differences in decay dynamics between time-resolved photoluminescence and transient absorption, results primarily from hole trapping in both NPs and sandwich NPs.

Charge Carriers Modulate the Bonding of Semiconductor Nanoparticle Dopants As Revealed by Time-Resolved X-ray Spectroscopy.

Understanding the electronic structure of doped semiconductors is essential to realize advancements in electronics and in the rational design of nanoscale devices. Reported here are the results of time-resolved X-ray absorption studies on copper-doped cadmium sulfide nanoparticles that provide an explicit description of the electronic dynamics of the dopants. The interaction of a dopant ion and an excess charge carrier is unambiguously observed via monitoring the oxidation state. The experimental data combined with DFT calculations demonstrate that dopant bonding to the host matrix is modulated by its interaction with charge carriers. Furthermore, the transient photoluminescence and the kinetics of dopant oxidation reveal the presence of two types of surface-bound ions that create midgap states.

Transient Melting and Recrystallization of Semiconductor Nanocrystals Under Multiple Electron-Hole Pair Excitation.

Ultrafast optical pump, X-ray diffraction probe experiments were performed on CdSe nanocrystal (NC) colloidal dispersions as functions of particle size, polytype, and pump fluence. Bragg peak shifts related to heating and peak amplitude reduction associated with lattice disordering are observed. For smaller NCs, melting initiates upon absorption of as few as ∼15 electron-hole pair excitations per NC on average (0.89 excitations/nm3 for a 1.5 nm radius) with roughly the same excitation density inducing melting for all examined NCs. Diffraction intensity recovery kinetics, attributable to recrystallization, occur over hundreds of picoseconds with slower recoveries for larger particles. Zincblende and wurtzite NCs revert to initial structures following intense photoexcitation suggesting melting occurs primarily at the surface, as supported by simulations. Electronic structure calculations relate significant band gap narrowing with decreased crystallinity. These findings reflect the need to consider the physical stability of nanomaterials and related electronic impacts in high intensity excitation applications such as lasing and solid-state lighting.

Nanomaterial-based sensors for the detection of biological threat agents.

The danger posed by biological threat agents and the limitations of modern detection methods to rapidly identify them underpins the need for continued development of novel sensors. The application of nanomaterials to this problem in recent years has proven especially advantageous. By capitalizing on large surface/volume ratios, dispersability, beneficial physical and chemical properties, and unique nanoscale interactions, nanomaterial-based biosensors are being developed with sensitivity and accuracy that are starting to surpass traditional biothreat detection methods, yet do so with reduced sample volume, preparation time, and assay cost. In this review, we start with an overview of bioagents and then highlight the breadth of nanoscale sensors that have recently emerged for their detection.

Giant optical enhancement of strain gradient in ferroelectric BiFeO3 thin films and its physical origin.

Through mapping of the spatiotemporal strain profile in ferroelectric BiFeO3 epitaxial thin films, we report an optically initiated dynamic enhancement of the strain gradient of 10(5)-10(6) m(-1) that lasts up to a few ns depending on the film thickness. Correlating with transient optical absorption measurements, the enhancement of the strain gradient is attributed to a piezoelectric effect driven by a transient screening field mediated by excitons. These findings not only demonstrate a new possible way of controlling the flexoelectric effect, but also reveal the important role of exciton dynamics in photostriction and photovoltaic effects in ferroelectrics.

Fast, Ratiometric FRET from Quantum Dot Conjugated Stabilized Single Chain Variable Fragments for Quantitative Botulinum Neurotoxin Sensing.

Botulinum neurotoxin (BoNT) presents a significant hazard under numerous realistic scenarios. The standard detection scheme for this fast-acting toxin is a lab-based mouse lethality assay that is sensitive and specific, but slow (∼2 days) and requires expert administration. As such, numerous efforts have aimed to decrease analysis time and reduce complexity. Here, we describe a sensitive ratiometric fluorescence resonance energy transfer scheme that utilizes highly photostable semiconductor quantum dot (QD) energy donors and chromophore conjugation to compact, single chain variable antibody fragments (scFvs) to yield a fast, fieldable sensor for BoNT with a 20-40 pM detection limit, toxin quantification, adjustable dynamic range, sensitivity in the presence of interferents, and sensing times as fast as 5 min. Through a combination of mutations, we achieve stabilized scFv denaturation temperatures of more than 60 °C, which bolsters fieldability. We also describe adaptation of the assay into a microarray format that offers persistent monitoring, reuse, and multiplexing.

Electric Field Modulation of Semiconductor Quantum Dot Photoluminescence: Insights Into the Design of Robust Voltage-Sensitive Cellular Imaging Probes.

The intrinsic properties of quantum dots (QDs) and the growing ability to interface them controllably with living cells has far-reaching potential applications in probing cellular processes such as membrane action potential. We demonstrate that an electric field typical of those found in neuronal membranes results in suppression of the QD photoluminescence (PL) and, for the first time, that QD PL is able to track the action potential profile of a firing neuron with millisecond time resolution. This effect is shown to be connected with electric-field-driven QD ionization and consequent QD PL quenching, in contradiction with conventional wisdom that suppression of the QD PL is attributable to the quantum confined Stark effect.

Synthesis and Ligand Exchange of Thiol-Capped Silicon Nanocrystals.

Hydride-terminated silicon (Si) nanocrystals were capped with dodecanethiol by a thermally promoted thiolation reaction. Under an inert atmosphere, the thiol-capped nanocrystals exhibit photoluminescence (PL) properties similar to those of alkene-capped Si nanocrystals, including size-tunable emission wavelength, relatively high quantum yields (>10%), and long radiative lifetimes (26-280 µs). X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy confirmed that the ligands attach to the nanocrystal surface via covalent Si-S bonds. The thiol-capping layer, however, readily undergoes hydrolysis and severe degradation in the presence of moisture. Dodecanethiol could be exchanged with dodecene by hydrosilylation for enhanced stability.

Picosecond energy transfer and multiexciton transfer outpaces Auger recombination in binary CdSe nanoplatelet solids.

Fluorescence resonance energy transfer (FRET) enables photosynthetic light harvesting, wavelength downconversion in light-emitting diodes (LEDs), and optical biosensing schemes. The rate and efficiency of this donor to acceptor transfer of excitation between chromophores dictates the utility of FRET and can unlock new device operation motifs including quantum-funnel solar cells, non-contact chromophore pumping from a proximal LED, and markedly reduced gain thresholds. However, the fastest reported FRET time constants involving spherical quantum dots (0.12-1 ns; refs 7-9) do not outpace biexciton Auger recombination (0.01-0.1 ns; ref. 10), which impedes multiexciton-driven applications including electrically pumped lasers and carrier-multiplication-enhanced photovoltaics. Few-monolayer-thick semiconductor nanoplatelets (NPLs) with tens-of-nanometre lateral dimensions exhibit intense optical transitions and hundreds-of-picosecond Auger recombination, but heretofore lack FRET characterizations. We examine binary CdSe NPL solids and show that interplate FRET (∼6-23 ps, presumably for co-facial arrangements) can occur 15-50 times faster than Auger recombination and demonstrate multiexcitonic FRET, making such materials ideal candidates for advanced technologies.

Intracellular FRET-based probes: a review.

Probes that exploit Förster resonance energy transfer (FRET) in their feedback mechanism are touted for their sensitivity, robustness, and low background, and thanks to the exceptional distance dependence of the energy transfer process, they provide a means of probing lengthscales well below the resolution of light. These attributes make FRET-based probes superbly suited to an intracellular environment, and recent developments in biofunctionalization and expansion of imaging capabilities have put them at the forefront of intracellular studies. Here, we present an overview of the engineering and execution of a variety of recent intracellular FRET probes, highlighting the diversity of this class of materials and the breadth of application they have found in the intracellular environment.

Silicon nanocrystals at elevated temperatures: retention of photoluminescence and diamond silicon to ß-silicon carbide phase transition.

We report the photoluminescence (PL) properties of colloidal Si nanocrystals (NCs) up to 800 K and observe PL retention on par with core/shell structures of other compositions. These alkane-terminated Si NCs even emit at temperatures well above previously reported melting points for oxide-embedded particles. Using selected area electron diffraction (SAED), powder X-ray diffraction (XRD), liquid drop theory, and molecular dynamics (MD) simulations, we show that melting does not play a role at the temperatures explored experimentally in PL, and we observe a phase change to ß-SiC in the presence of an electron beam. Loss of diffraction peaks (melting) with recovery of diamond-phase silicon upon cooling is observed under inert atmosphere by XRD. We further show that surface passivation by covalently bound ligands endures the experimental temperatures. These findings point to covalently bound organic ligands as a route to the development of NCs for use in high temperature applications, including concentrated solar cells and electrical lighting.

Near-infrared photoluminescence enhancement in Ge/CdS and Ge/ZnS Core/shell nanocrystals: utilizing IV/II-VI semiconductor epitaxy.

Ge nanocrystals have a large Bohr radius and a small, size-tunable band gap that may engender direct character via strain or doping. Colloidal Ge nanocrystals are particularly interesting in the development of near-infrared materials for applications in bioimaging, telecommunications and energy conversion. Epitaxial growth of a passivating shell is a common strategy employed in the synthesis of highly luminescent II-VI, III-V and IV-VI semiconductor quantum dots. Here, we use relatively unexplored IV/II-VI epitaxy as a way to enhance the photoluminescence and improve the optical stability of colloidal Ge nanocrystals. Selected on the basis of their relatively small lattice mismatch compared with crystalline Ge, we explore the growth of epitaxial CdS and ZnS shells using the successive ion layer adsorption and reaction method. Powder X-ray diffraction and electron microscopy techniques, including energy dispersive X-ray spectroscopy and selected area electron diffraction, clearly show the controllable growth of as many as 20 epitaxial monolayers of CdS atop Ge cores. In contrast, Ge etching and/or replacement by ZnS result in relatively small Ge/ZnS nanocrystals. The presence of an epitaxial II-VI shell greatly enhances the near-infrared photoluminescence and improves the photoluminescence stability of Ge. Ge/II-VI nanocrystals are reproducibly 1-3 orders of magnitude brighter than the brightest Ge cores. Ge/4.9CdS core/shells show the highest photoluminescence quantum yield and longest radiative recombination lifetime. Thiol ligand exchange easily results in near-infrared active, water-soluble Ge/II-VI nanocrystals. We expect this synthetic IV/II-VI epitaxial approach will lead to further studies into the optoelectronic behavior and practical applications of Si and Ge-based nanomaterials.

In situ optical and structural studies on photoluminesence quenching in CdSe/CdS/Au heterostructures.

We report here detailed in situ studies of nucleation and growth of Au on CdSe/CdS nanorods using synchrotron SAXS technique and time-resolved spectroscopy. We examine structural and optical properties of CdSe/CdS/Au heterostructures formed under UV illumination. We compare the results for CdSe/CdS/Au heterostructures with the results of control experiments on CdSe/CdS nanorods exposed to gold precursor under conditions when no such heterostructures are formed (no UV illumination). Our data indicate similar photoluminescence (PL) quenching and PL decay profiles in both types of samples. Via transient absorption and PL, we show that such behavior is consistent with rapid (faster than 3 ps) hole trapping by gold-sulfur sites at the surface of semiconductor nanoparticles. This dominant process was overlooked in previous end-point studies on semiconductor/metal heterostructures.

Thermal stability of colloidal InP nanocrystals: small inorganic ligands boost high-temperature photoluminescence.

We examine the stability of excitons in quantum-confined InP nanocrystals as a function of temperature elevation up to 800 K. Through the use of static and time-resolved spectroscopy, we find that small inorganic capping ligands substantially improve the temperature dependent photoluminescence quantum yield relative to native organic ligands and perform similarly to a wide band gap inorganic shell. For this composition, we identify the primary exciton loss mechanism as electron trapping through a combination of transient absorption and transient photoluminescence measurements. Density functional theory indicates little impact of studied inorganic ligands on InP core states, suggesting that reduced thermal degradation relative to organic ligands yields improved stability; this is further supported by a lack of size dependence in photoluminescence quenching, pointing to the dominance of surface processes, and by relative thermal stabilities of the surface passivating media. Thus, small inorganic ligands, which benefit device applications due to improved carrier access, also improve the electronic integrity of the material during elevated temperature operation and subsequent to high temperature material processing.

Tetraalkylammonium uranyl isothiocyanates.

Three tetraalkylammonium uranyl isothiocyanates, [(CH(3))(4)N](3)UO(2)(NCS)(5) (1), [(C(2)H(5))(4)N](3)UO(2)(NCS)(5) (2), and [(C(3)H(7))(4)N](3)UO(2)(NCS)(5) (3), have been synthesized from aqueous solution and their structures determined by single-crystal X-ray diffraction. All of the compounds consist of the uranyl cation equatorially coordinated to five N-bound thiocyanate ligands, UO(2)(NCS)(5)(3-), and charge-balanced by three tetraalkylammonium cations. Raman spectroscopy data have been collected on compounds 1-3, as well as on solutions of uranyl nitrate with increasing levels of sodium thiocyanate. By tracking the Raman signatures of thiocyanate, the presence of both free and bound thiocyanate is confirmed in solution. The shift in the Raman signal of the uranyl symmetric stretching mode suggests the formation of higher-order uranyl thiocyanate complexes in solution, while the solid-state Raman data support homoleptic isothiocyanate coordination about the uranyl cation. Presented here are the syntheses and crystal structures of 1-3, pertinent Raman spectra, and a discussion regarding the relationship of these isothiocyanates to previously described uranyl halide phases, UO(2)X(4)(2-).

Uranyl sensitization of samarium(III) luminescence in a two-dimensional coordination polymer.

Heterometallic carboxyphosphonates UO(2)(2+)/Ln(3+) have been prepared from the hydrothermal reaction of uranyl nitrate, lanthanide nitrate (Ln = Sm, Tb, Er, Yb), and phosphonoacetic acid (H(3)PPA). Compound 1, (UO(2))(2)(PPA)(HPPA)(2)Sm(H(2)O)·2H(2)O (1) adopts a two-dimensional structure in which the UO(2)(2+) metal ions bind exclusively to the phosphonate moiety, whereas the Ln(3+) ions are coordinated by both phosphonate and carboxylate functionalities. Luminescence studies of 1 show very bright visible and near-IR samarium(III)-centered emission upon direct excitation of the uranyl moiety. The Sm(3+) emissive state exhibits a double-exponential decay with lifetimes of 67.2 ± 6.5 and 9.0 ± 1.3 µs as measured at 594 nm, after excitation at both 365 and 420 nm. No emission is observed in the region typical of the uranyl cation, indicating that all energy is either transferred to the Sm(3+) center or lost to nonradiative processes. Herein we report the synthesis, crystal structure, and luminescent behavior of 1, as well as those of the isostructural terbium, erbium, and ytterbium analogues.

Hydrothermal synthesis of uranyl squarates and squarate-oxalates: hydrolysis trends and in situ oxalate formation.

We report the synthesis of two uranyl squarates and two mixed-ligand uranyl squarate-oxalates from aqueous solutions under hydrothermal conditions. These products exhibit a range of uranyl building units from squarates with monomers in (UO(2))(2)(C(4)O(4))(5).6NH(4).4H(2)O (1; a = 16.731(17) A, b = 7.280(8) A, c = 15.872(16) A, beta = 113.294(16) degrees , monoclinic, P2(1)/c) and chains in (UO(2))(2)(OH)(2)(H(2)O)(2)(C(4)O(4)) (2; a = 12.909(5) A, b = 8.400(3) A, c = 10.322(4) A, beta = 100.056(7) degrees , monoclinic, C2/c) to two squarate-oxalate polymorphs with dimers in (UO(2))(2)(OH)(C(4)O(4))(C(2)O(4)).NH(4).H(2)O (3; a = 9.0601(7) A, b = 15.7299(12) A, c = 10.5108(8) A, beta = 106.394(1) degrees , monoclinic, P2(1)/n; and 4; a = 8.4469(6) A, b = 7.7589(5) A, c = 10.5257(7) A, beta = 105.696(1) degrees , monoclinic, P2(1)/m). The dominance at low pH of monomeric species and the increasing occurrence of oligomeric species with increasing pH suggests that uranyl hydrolysis, mUO(2)(2+) + nH(2)O right harpoon over left harpoon [(UO(2))(m)(OH)(n)](2m-n) + nH(+), has a significant role in the identity of the inorganic building unit. Additional factors that influence product assembly include in situ hydrolysis of squaric acid to oxalic acid, dynamic metal to ligand concentration, and additional binding modes resulting from the introduction of oxalate anions. These points and the effects of uranyl hydrolysis with changing pH are discussed in the context of the compounds presented herein.

Capturing hydrolysis products in the solid state: effects of pH on uranyl squarates under ambient conditions.

Two uranyl squarates, (UO(2))(6)(C(4)O(4))(3)(OH)(6)O(2)·9H(2)O·4NH(4) (1; a = 16.6897(7) Å, cubic, I23) and (UO(2))(C(4)O(4))(OH)(2)·2NH(4) (2; a = 8.5151(4), b = 15.6822(8), c = 7.3974, orthorhombic, Pbcm), have been synthesized from ambient aqueous solutions as a function of pH. Oligomerization of the uranyl cation from monomeric pentagonal bipyramids (pH < 5) to [(UO(2))(3)O(OH)(3)] trimers (5 < pH < 8) in 1 and ultimately [(UO(2))(OH)(2)](n) chains (7 < pH < 8) in 2 is observed. This evolution of speciation versus pH is consistent with what has been observed in solution and thus may be represented by the uranyl hydrolysis equilibrium, mUO(2)(2+) + nH(2)O ↔ [(UO(2))(m)(OH)(n)](2m - n) + nH(+). Structural systematics, physical properties, and a discussion of species selectivity by squarate anions are presented.