Autonomous, broad-spectrum detection of hazardous aerosols in seconds

Actual or surrogate chemical, biological, radiological, nuclear, and explosive materials and illicit drug precursors can be rapidly detected and identified when in aerosol form by a Single-Particle Aerosol Mass Spectrometry (SPAMS) system. This entails not only the sampling of such particles but also the physical analysis and subsequent data analysis leading to a highly reliable alarm state. SPAMS hardware is briefly reviewed. SPAMS software algorithms are discussed in greater detail. A laboratory experiment involving actual threat and surrogate releases mixed with ambient background aerosols demonstrates broad-spectrum detection within seconds. Data from a field test at the San Francisco International Airport demonstrate extended field operation with an ultralow false alarm rate. Together these data sets demonstrate a significant and important advance in rapid aerosol threat detection.     

Simultaneous laser-induced fluorescence and scattering detection of individual particles separated by capillary electrophoresis

This technical note describes a detector capable of simultaneously monitoring scattering and fluorescence signals of individual particles separated by capillary electrophoresis. Due to its nonselective nature, scattering alone is not sufficient to identify analyte particles. However, when the analyte particles are fluorescent, the detector described here is able to identify simultaneously occurring scattering and fluorescent signals, even when contaminating particles (i.e., nonfluorescent) are present. Both fluorescent polystyrene particles and 10-nonyl acridine orange (NAO)-labeled mitochondria were used as models. Fluorescence versus scattering (FVS) plots made it possible to identify two types of particles and a contaminant in a mixture of polystyrene particles. We also analyzed NAO-labeled mitochondria before and after cryogenic storage; the mitochondria FVS plots changed with storage, which suggests that the detector reported here is suitable for monitoring subtle changes in mitochondrial morphology that would not be revealed by monitoring only fluorescence or scattering signals.     

Conditional-sampling spectrograph detection system for fluorescence measurements of individual airborne biological particles

We report the design and operation of a prototype conditional-sampling spectrograph detection system that can record the fluorescence spectra of individual, micrometer-sized aerosols as they traverse an intense 488-nm intracavity laser beam. The instrument's image-intensified CCD detector is gated by elastic scattering or by undispersed fluorescence from particles that enter the spectrograph's field of view. It records spectra only from particles with preselected scattering-fluorescence levels (a fiber-optic-photomultiplier subsystem provides the gating signal). This conditional-sampling procedure reduces data-handling rates and increases the signal-to-noise ratio by restricting the system's exposures to brief periods when aerosols traverse the beam. We demonstrate these advantages by reliably capturing spectra from individual fluorescent microspheres dispersed in an airstream. The conditional-sampling procedure also permits some discrimination among different types of particles, so that spectra may be recorded from the few interesting particles present in a cloud of background aerosol. We demonstrate such discrimination by measuring spectra from selected fluorescent microspheres in a mixture of two types of microspheres, and from bacterial spores in a mixture of spores and nonfluorescent kaolin particles.     

Reagentless mediated laccase electrode for the detection of enzyme modulators

We have investigated aerobic mediation of electron transfer to a laccase enzyme by the solution redox couples [Os(bpy)(2)Cl(2)](+/0) and [Os(bpy)(2)(MeIm)Cl](2+/+), where bpy is 2,2-bipyridine and MeIm is N-methylimidazole. The factors that influence the homogeneous mediation reaction are investigated and discussed. Investigation of ionic strength, pH, and temperature effects on the kinetics of intermolecular electron transfer elucidates the governing factors in the mediator-enzyme reactions. Coimmobilization of both enzyme and an osmium redox mediator in a hydrogel on glassy carbon electrodes results in a biosensor for the reagentless addressing of enzyme activity, consuming only oxygen present in solution. Thus, these immobilized enzyme biosensors can be utilized for the detection of modulators of laccase activity, such as the inhibitor sodium azide. The enzyme inhibition biosensor can detect levels of azide as low as 2.5 × 10(-6) mol dm(-3) in solution.     

Reagentless detection of Mycobacteria tuberculosis H37Ra in respiratory effluents in minutes

Two similar mycobacteria, Mycobacteria tuberculosis H37Ra and Mycobacteria smegmatis are rapidly detected and identified within samples containing a complex background of respiratory effluents using single-particle aerosol mass spectrometry (SPAMS). M. tuberculosis H37Ra (TBa), an avirulent strain, is used as a surrogate for virulent tuberculosis; M. smegmatis (MSm) is utilized as a near-neighbor confounder for TBa. Bovine lung surfactant and human exhaled breath condensate are used as first-order surrogates for infected human lung expirations from patients with pulmonary tuberculosis. This simulated background sputum is mixed with TBa or MSm and nebulized to produce conglomerate aerosol particles, single particles that contain a bacterium embedded within a background respiratory matrix. Mass spectra of single conglomerate particles exhibit ions associated with both respiratory effluents and mycobacteria. Spectral features distinguishing TBa from MSm in pure and conglomerate particles are shown. SPAMS pattern matching alarm algorithms are able to distinguish TBa-containing particles from background matrix and MSm for >50% of the test particles, which is sufficient to enable a high probability of detection and a low false alarm rate if an adequate number of such particles are present. These results indicate the potential usefulness of SPAMS for rapid, reagentless tuberculosis screening.     

Laser power dependence of mass spectral signatures from individual bacterial spores in bioaerosol mass spectrometry

Bioaerosol mass spectrometry is being developed to analyze and identify biological aerosols in real time. Characteristic mass spectra from individual bacterial endospores of Bacillus subtilis var. niger were obtained in a bipolar aerosol time-of-flight mass spectrometer using a pulsed 266-nm laser for molecular desorption and ionization. Spectra from single spores collected at an average fluence of approximately 0.1 J/cm2 frequently contain prominent peaks attributed to arginine, dipicolinic acid, and glutamic acid, but the shot-to-shot (spore-to-spore) variability in the data may make it difficult to consistently distinguish closely related Bacillus species with an automated routine. Fortunately, a study of the laser power dependence of the mass spectra reveals clear trends and a finite number of "spectral types" that span most of the variability. This, we will show, indicates that a significant fraction of the variability must be attributed to fluence variations in the profile of the laser beam.     

Compression properties of individual micron-sized acrylic particles

Compression behaviours of individual micron-sized acrylic particles have been studied by using a nanoindentation-based method with a flat-end tip, instead of a commonly used sharp tip. The effect of loading rate on the mechanical properties of the high monodisperse acrylic particles, such as K-value, breaking force and breaking displacement, has been investigated. A wide range of loading rate conditions has been applied to examine the sensitivity of selected mechanical properties. It has been observed that the loading rate has a significant effect on K-value and breaking force whereas breaking displacement is independent of loading rate. These results facilitate the industrial applications of acrylic particles.     

Detection of pesticide residues on individual particles

An aerosol time-of-flight mass spectrometer (ATOFMS) is used to analyze the size and composition of individual particles containing pesticides. Pesticide residues are found in the atmosphere as a result of spray drift, volatilization, and suspension of coated soils. The ability of the ATOFMS to identify the presence of these contaminants on individual particles is assessed for particles created from pure solutions of several commonly used pesticides, as well as pesticides mixed with an organic matrix, and coated on soils. The common names of the pesticides studied are 2,4-D, atrazine, chlorpyrifos, malathion, permethrin, and propoxur. Analysis of the mass spectra produced by single- and two-step laser desorption/ionization of pesticide-containing particles allows for identification of peaks that can be used for detection of pesticide residues in the ambient aerosol. The identified marker peaks are used to approximate detection limits for the pesticides applied to soils, which are on the order of a fraction of a monolayer for individual particles. Results suggest that this technique may be useful for studying the real-time partitioning and distribution of pesticides in the atmosphere immediately following application in agricultural regions.     

Achieving high detection sensitivity (14 zmol) of biomolecular ions in bioaerosol mass spectrometry

Bioaerosol mass spectrometry (BAMS) performs single-cell analysis in real time. However, the specificity of BAMS mass signatures has been limited by low sensitivity at high masses. To increase the mass range and sensitivity of BAMS, a novel design was developed that utilizes a linear flight tube with delayed extraction and an electrostatic ion guide. This study quantifies the sensitivity limits of the novel BAMS design and evaluates the feasibility of BAMS to detect higher mass biomarkers from single cells. All experiments were carried out using MALDI aerosol particles that were nebulized from solution. Sensitivity was assessed by generating particles with decreasing amounts of analyte via serial dilutions. The amount of analyte contained within each particle was calculated based on particle size, density, and molarity of the analyte within solution. A variety of biomolecular ions were studied and signals obtained from particles containing 300 zmol of maltopentaose, 132 zmol of alpha-cyclodextrin, and 14 zmol (approximately 8400 molecules) of gramicidin S are reported. The detection of 14 zmol of gramicidin S is to the best of our knowledge a record in sensitivity for MALDI TOF-MS.     

Measurement of rotation of individual spherical particles in cohesive granulates

To explore dynamical processes in granular matter, we use a combination of 3D imaging and mechanical testing. We analyze structural changes using confocal microscopy while applying a compression load simultaneously. Fluorescently labeled polydisperse silica particles were hydrophobized with long alkyl chains and dispersed in an index-matching liquid. The particles show a weak attraction. Photobleaching the central plane of individual particles generates an optical anisotropy without changing particle interaction. In a series of 3D images, we follow trajectories and rotation of single particles. We focus on particle translation and rotation in dependency of the local volume fraction. During compression, restructuring happens predominantly in regions of low packing density. We show that rotation plays an important role and is hence a key parameter for explaining dynamical processes in granular systems.     

Detection of negative ions from individual ultrafine particles

Aerosol mass spectrometers can be used to classify individual airborne particles on the basis of chemical composition. While positive ion mass spectra are normally used to characterize ultrafine particles (defined here as particles smaller than 200 nm in diameter), negative ion mass spectra can provide complementary information. To effectively utilize the negative ion mass spectra of ultrafine particles, it is important to understand biases in the formation and detection of negative ions. It is found that the intensity of negative ions is generally less than that of positive ions, due to the creation of electrons in the ablation process that must react to form negative ions. The ablation efficiency, defined as the probability that an ablated particle produces a detectable ion signal, exhibits both size and composition dependencies. The ablation efficiency for detection of negative ions follows the same trends as the ablation efficiency for the detection of positive ions: sodium chloride and ammonium nitrate have higher ablation efficiencies than oleic acid, and the ablation efficiency decreases with the particle diameter. The ablation efficiency of negative ions is less than or equal to the ablation efficiency of positive ions, and the relative difference increases as the particle diameter decreases. Pure ammonium sulfate particles exhibit an ablation efficiency too low to be measured in the present experiments. However, trace amounts of sulfate in mixed-composition particles can be readily detected in the negative ion mass spectra.     

Investigation of the utility of laser-secondary neutral mass spectrometry for the detection of polyaromatic hydrocarbons in individual atmospheric aerosol particles

The distribution of polyaromatic hydrocarbons (PAHs) in ambient aerosol particles is of importance to both human health and climate forcing. Although time-of-flight secondary ion mass spectrometry (ToF-SIMS) has proven useful for studying the distribution of organic compounds in individual aerosol particles, it is difficult to detect PAHs at relevant concentrations in individual aerosol particles because of their low ion yield. In this study, we explore the potential of using laser secondary neutral mass spectrometry (Laser-SNMS) to study three PAHs: pyrene, anthracene, and naphthalene. Because of the high volatility of PAHs, a cryostage was required for the analysis to prevent sublimation of the molecules into the vacuum chamber. We studied two laser systems, a 157 nm excimer laser, which is capable of single-photon ionization of the PAHs, and a 193 nm laser, which requires multiphoton ionization. Under optimized conditions for laser power density and primary ion pulse length, 193 nm postionization resulted in a 2-50-fold increase in ion yield over ToF-SIMS. Using the 157 nm laser, the yield was increased by more than 3 orders of magnitude for all 3 PAHs studied. The single-photon postionization process proved superior in terms of both yield enhancement and reduced fragmentation. By using the optimized 157 nm laser system and a cryostage, we were able to detect PAHs on the surface of 2 μm diameter ambient aerosol particles.     

Electro-elastic tuning of single particles in individual self-assembled quantum dots

We investigate the effect of uniaxial stress on InGaAs quantum dots in a charge tunable device. Using Coulomb blockade and photoluminescence, we observe that significant tuning of single particle energies (≈-0.22 meV/MPa) leads to variable tuning of exciton energies (+18 to -0.9 μeV/MPa) under tensile stress. Modest tuning of the permanent dipole, Coulomb interaction and fine-structure splitting energies is also measured. We exploit the variable exciton response to tune multiple quantum dots on the same chip into resonance.     

Comprehensive assignment of mass spectral signatures from individual Bacillus atrophaeus spores in matrix-free laser desorption/ionization bioaerosol mass spectrometry

We have fully characterized the mass spectral signatures of individual Bacillus atrophaeus spores obtained using matrix-free laser desorption/ionization bioaerosol mass spectrometry (BAMS). Mass spectra of spores grown in unlabeled, 13C-labeled, and 15N-labeled growth media were used to determine the number of carbon and nitrogen atoms associated with each mass peak observed in mass spectra from positive and negative ions. To determine the parent ion structure associated with fragment ion peaks, the fragmentation patterns of several chemical standards were independently determined. Our results confirm prior assignments of dipicolinic acid, amino acids, and calcium complex ions made in the spore mass spectra. The identities of several previously unidentified mass peaks, key to the recognition of Bacillus spores by BAMS, have also been revealed. Specifically, a set of fragment peaks in the negative polarity is shown to be consistent with the fragmentation pattern of purine nucleobase-containing compounds. The identity of m/z = +74, a marker peak that helps discriminate B. atrophaeus from Bacillus thuringiensis spores grown in rich media is [N1C4H12]+. A probable precursor molecule for the [N1C4H12]+ ion observed in spore spectra is trimethylglycine (+N(CH3)3CH2COOH), which produces a m/z = +74 peak when ionized in the presence of dipicolinic acid. A clear assignment of all the mass peaks in the spectra from bacterial spores, as presented in this work, establishes their relationship to the spore chemical composition and facilitates the evaluation of the robustness of "marker" peaks. This is especially relevant for peaks that have been used to discriminate Bacillus spore species, B. thuringiensis and B. atrophaeus, in our previous studies.     

Mathematical modeling of thermal processing of individual solid-fuel particles

A mathematical model, an algorithm, and a program for calculating the thermal processing of individual solid-fuel particles are developed with account for moisture evaporation, escape of volatiles, and burn-out of the carbon residue. Numerical calculations of the influence of regime conditions on the gasification-combustion of individual particles of Chelyabinsk brown coal are performed. A comparison with experiment is made.Institute for Problems of Energy Conservation, Academy of Sciences of Ukraine, Branch of High-Temperature Energy Conversion, Kiev. Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 68, No. 1, pp. 96–109, January–February, 1995.     

Material-specific detection and classification of single nanoparticles

Detection and classification of nanoparticles are important for environmental monitoring, contamination mitigation, biological label tracking, and biodefense. Detection techniques involve a trade-off between sensitivity, discrimination, and speed. This paper presents a material-specific dual-color common-path interferometric detection system. Two wavelengths are simultaneously used to discriminate between 60 nm silver and 80 nm diameter gold particles in solution with a detection time of τ ≈ 1 ms. The detection technique is applicable to situations where both particle size and material are of interest.     

Determination of low-Z elements in individual environmental particles using windowless EPMA

The determination of low-Z elements such as carbon, nitrogen, and oxygen in atmospheric aerosol particles is of interest in studying environmental pollution. Conventional electron probe microanalysis technique has a limitation for the determination of the low-Z elements, mainly because the Be window in an energy-dispersive X-ray (EDX) detector hinders the detection of characteristic X-rays from light elements. The feasibility of low-Z element determination in individual particles using a windowless EDX detector is investigated. To develop a method capable of identifying chemical species of individual particles, both the matrix and the geometric effects of particles have to be evaluated. X-rays of low-Z elements generated by an electron beam are so soft that important matrix effects, mostly due to X-ray absorption, exist even within particles in the micrometer size range. Also, the observed radiation, especially that of light elements, experiences different extents of absorption, depending on the shape and size of the particles. Monte Carlo calculation is applied to explain the variation of observed X-ray intensities according to the geometric and chemical compositional variation of individual particles, at different primary electron beam energies. A comparison is carried out between simulated and experimental data, collected for standard individual particles with chemical compositions as generally observed in marine and continental aerosols. Despite the many fundamental problematic analytical factors involved in the observation of X-rays from low-Z elements, the Monte Carlo calculation proves to be quite reliable to evaluate those matrix and geometric effects. Practical aspects of the Monte Carlo calculation for the determination of light elements in individual particles are also considered.     

Spatially correlated fluorescence/AFM of individual nanosized particles and biomolecules

Individual fluorescent polystyrene nanospheres (<10-100-nm diameter) and individual fluorescently labeled DNA molecules were dispersed on mica and analyzed using time-resolved fluorescence spectroscopy and atomic force microscopy (AFM). Spatial correlation of the fluorescence and AFM measurements was accomplished by (1) positioning a single fluorescent particle into the near diffraction-limited confocal excitation region of the optical microscope, (2) recording the time-resolved fluorescence emission, and (3) measuring the intensity of the excitation laser light scattered from the apex of an AFM probe tip and the AFM topography as a function of the lateral position of the tip relative to the sample substrate. The latter measurements resulted in concurrent high-resolution (approximately 10-20 nm laterally) images of the laser excitation profile of the confocal microscope and the topography of the sample. Superposition of these optical and topographical images enabled unambiguous identification of the sample topography residing within the excitation region of the optical microscope, facilitating the identification and structural characterization of the nanoparticle(s) or biomolecule(s) responsible for the fluorescence signal observed in step 2. These measurements also provided the lateral position of the particles relative to the laser excitation profile and the surrounding topography with nanometer-scale precision and the relationship between the spectroscopic and structural properties of the particles. Extension of these methods to the study of other types of nanostructured materials is discussed.     

Pyrometric measurement of the temperature and size of individual combusting fuel particles

A two-color pyrometric technique was developed for the in situ simultaneous measurement of individual fuel-particle temperatures and sizes in a pressurized entrained flow reactor (PEFR). A method that requires only a single optical port was developed, and a specially designed optical probe was manufactured for the measurements at the PEFR. The fuel-particle temperature was obtained by applying two-color pyrometry. The particle size was determined from the same pyrometric signals. A discrimination method was developed to confirm that the detected particle was valid for particle sizing. Several series of measurements were made at a PEFR at different process conditions, and some typical results are shown.     

Direct detection of Dark Matter particles

Arguments on the investigation of the Dark Matter particles in the galactic halo are addressed. Recent results obtained by the DAMA/LIBRA set-up—exploiting the model independent annual modulation signature for Dark Matter (DM) particles—are shortly summarized. In fact, the DAMA project is an observatory for rare processes and it is operative deep underground at the Gran Sasso National Laboratory of the INFN. Its main apparatus is at present the DAMA/LIBRA set-up, whose sensitive volume is made of highly radiopure NaI(Tl) detectors. The first DAMA/LIBRA results confirm the evidence for the presence of DM particles in the galactic halo as pointed out by the former DAMA/NaI set-up; cumulatively the data support such evidence at 8.2σ CL and satisfy all the many peculiarities of the DM annual modulation signature. Future perspectives are briefly addressed.