Measurement and autocorrelation analysis of two-dimensional light-scattering patterns from living cells for label-free classification.

We incorporate optics and an ICCD to record the two-dimensional angular optical scattering (TAOS) patterns retrieved from single aerosolized cells. We analyze these patterns by performing autocorrelations and demonstrate that we are able to retrieve cell size from the locations of the secondary maxima. Additional morphological information is contained in the autocorrelation functions and decay rate of the heights of the autocorrelation peaks. We demonstrate these techniques with C6 and Y79 cells, which are readily distinguishable. One key advantage of this methodology is that there is no requirement for antibody and fluorescent labeling molecules.

Fluorescence spectra of atmospheric aerosol particles measured using one or two excitation wavelengths: comparison of classification schemes employing different emission and scattering results.

An improved Dual-wavelength-excitation Particle Fluorescence Spectrometer (DPFS) has been reported. It measures two fluorescence spectra excited sequentially by lasers at 263 nm and 351 nm, from single atmospheric aerosol particles in the 1-10 mum diameter size range. Here we investigate the different levels of discrimination capability obtained when different numbers of excitation and fluorescence-emission wavelengths are used for analysis. We a) use the DPFS to measure fluorescence spectra of Bacillus subtilis and other aerosol particles, and a 25-hour sample of atmospheric aerosol at an urban site in Maryland, USA; b) analyze the data using six different algorithms that employ different levels of detail of the measured data; and c) show that when more of the data measured by the DPFS is used, the ability to discriminate among particle types is significantly increased.

Particle-fluorescence spectrometer for real-time single-particle measurements of atmospheric organic carbon and biological aerosol.

A particle-fluorescence spectrometer (PFS) for real-time measurements of single-particle UV-laser-induced fluorescence (UV-LIF) excited with a pulsed (263-nm) laser is reported. The dispersed UV-LIF spectra are measured by a 32-anode PMT detector with spectral coverage from 280-600 nm. The PFS represents a significant improvement over our previous apparatus [Pinnick et al., Atmos. Environ. 2004, 38, 1657] and can (1) measure fluorescence spectra of bacterial particles having light-scattering sizes as small as 1 microm (previously limited to about 3 microm) and so can measure particles with size in the range of 1-10 microm, (2) measure each particle's elastic scattering which can be used to estimate particle size (not available previously), (3) measure single-particle fluorescence spectra with a laser and detector that can record spectra as fast as 90,000/s, although the highest rates we have found experimentally in atmospheric measurements is only several hundred per second (previously limited by detectors to only 25/s), and (4) provide a time stamp for a data block of spectra with time resolution from 10 ms to 10 min. In addition, the PFS has been modified to be more robust, transportable, and smaller. The use of an aerodynamic-focusing sheath inlet nozzle assembly has improved the sample rate. The PFS has been employed to measure UV-LIF spectra from individual atmospheric particles during October-December 2006 and January-May 2008 in New Haven, CT, and during January-May 2007 in Las Cruces, NM.

Real-time measurement of dual-wavelength laser-induced fluorescence spectra of individual aerosol particles.

We report the development of an in-situ aerosol detection system capable of rapidly measuring dual-wavelength laser-induced fluorescence spectra of single particles on the fly using a single spectrometer and a single 32-anode photomultiplier array. We demonstrate the capability of this system with both reference samples and outdoor air. We present spectra from separate excitation wavelengths from the same particle that demonstrate improved discrimination capability compared with only using one excitation wavelength.

Wavelength and intensity switching in directly coupled semiconductor microdisk lasers.

We demonstrate output wavelength and intensity switching in a three-element directly coupled microdisk device consisting of one spiral microdisk coupled to two semicircle microdisks. The gapless coupling mechanism used allows individual elements to achieve lasing while achieving optimal transfer of optical power between adjacent microdisks. By controlling the transparency of the center element via injection current, the edge elements can be allowed to exchange their amplified spontaneous emission. In this manner, on-off-on switching of the output intensity, as well as discontinuous shifts in the output wavelength, can be achieved as a function of increasing injection current.

Linear and non-linear spectroscopy of microparticles: basic principles, new techniques and promising applications.

In the introduction a brief recollection is made of how one of us (RKC), accidentally, got into this field of linear and nonlinear spectroscopy of a dielectric micro-particle that can be treated as a micro-cavity or a micro-resonator. The basic principles of whispering gallery modes (WGMs) and their relationship with electromagnetic theory are presented. To simplify the mathematics, we only discuss an example from a 2-d case of light illumination perpendicular to the fiber axis. This 2-d example has relevance to semiconductor circular disk lasers, nonlinear optics in torroids, fibers and spheres at the tip of a fiber. The internal and near-field distribution of a WGM are graphically plotted to give the reader a chance to get a physical understanding of the spatial distribution as well as spectral distribution of WGMs. Several new techniques that enable the measurements of: (1) nanometer changes in the cladding diameter over a centimeter length of fiber; (2) some aspects of the morphology of micro-particles by elastic scattering; and (3) biochemical reactions at the interface of liquid media with a sphere at the end of a fiber. A few interesting nonlinear optical experimental results pertaining to stimulated Raman scattering (SRS) are touched upon. We present some preliminary results for promising applications in the area of bioaerosols. These include ambient aerosol characterization and identification with elastic scattering, fluorescence spectroscopy, and other optical and/or biochemical identifiers.

Angularly resolved light scattering from aerosolized spores: observations and calculations.

Angularly resolved elastic light scattering patterns from individual aerosolized Bacillus subtilis spores were qualitatively compared with simulations. Two-dimensional angular optical scattering patterns of the spores were collected for polar scattering angles varying from approximately 77 degrees to 130 degrees and azimuthal angles varying from 0 degrees to 360 degrees . Computations were performed with single T-matrix formalism by simulating a spore with three different particle shapes: (1) a finite-length cylinder with spherical end caps, (2) a spheroid, and (3) two spheres in contact. Excellent agreement between computation and measurement was found for the finite-length cylinder with spherical end caps, poorer agreement was found for the spheroids, and the poorest agreement was for the two spheres in contact.

Simultaneous forward- and backward-hemisphere elastic-light-scattering patterns of respirable-size aerosols.

Two-dimensional angular optical scattering (TAOS) patterns of aerosols are measured simultaneously from the forward hemisphere 15 degrees < theta < 90 degrees as well as the backward hemisphere 90 degrees < theta < 165 degrees (detecting 63% of the 4pi sr of scattered light) by using an ellipsoidal reflector and an intensified CCD detector. TAOS patterns were obtained from polystyrene-latex spheres (individuals and aggregates) and from single Bacillus subtilis spores. These information-rich patterns, measured with a single laser pulse for individual particles on the fly, suggest that forward-TAOS and backward-TAOS measurements may be used for rapid classification of single aerosol particles.

Optical scattering by biological aerosols: experimental and computational results on spore simulants.

We present both a computational and an experimental approach to the problem of biological aerosol characterization, joining the expertises reached in the field of theoretical optical scattering by complex, arbitrary shaped particles (multipole expansion of the electromagnetic fields and Transition Matrix), and a novel experimental technique based on two-dimensional angular optical scattering (TAOS). The good agreement between experimental and computational results, together with the possibility for a laboratory single-particle angle-resolved investigation, opens a new scenario in biological particle modelling, and might have major implications for a rapid discrimination of airborne particles.

Multivariate analysis and classification of two-dimensional angular optical scattering patterns from aggregates.

Two-dimensional light-scattering patterns from aggregates have undergone feature extraction followed by multivariate statistical analysis. The aggregates are comprised of primary particles of varying shape and size. Morphological descriptors (features) were extracted by a nonlinear filtering algorithm (spectrum enhancement) and then processed by principal component analysis and discriminant function analysis. The analysis was performed on two data sets, one in which the aggregates had a fixed primary particle size but varied in overall dimension and another in which the aggregate size was fixed but the primary particle size varied. Classification of the samples was performed adequately, providing some distinction among the limited classes that were analyzed.

Two-dimensional angular optical scattering patterns of microdroplets in the mid infrared with strong and weak absorption.

Two-dimensional angular optical scattering (TAOS) patterns of droplets composed of a mixture of H2O and D2O are detected in the mid infrared. First, a lens is used in the Abbé sine condition to collect a small solid angle of light, where the scattering pattern matches well numerical simulations based on Mie theory. Next, TAOS patterns from droplets spanning a large (approximately 27pi sr) solid angle are captured simultaneously at two wavelengths. The effects of absorption are evident in the patterns and are discernible without the need for curve matching by Mie theory.

Application of light-emitting diodes for aerosol fluorescence detection.

We demonstrate a proof-of-concept optical spectroscopic system for bioaerosol-particle fluorescence detection, in which a pulsed high-power laser is replaced by a highly compact linear array of sequentially fired light from blue light-emitting diodes. The results suggest that low-cost, compact optical aerosol detection may be feasible with the contemporary emergence of efficient UV light-emitting diodes.

Characterizing and monitoring respiratory aerosols by light scattering.

The elastic-scattering intensity pattern from a single particle as a function of spherical coordinate angles theta and phi provides detailed information on the pattern's morphology. By use of an ellipsoidal reflector and a CCD camera, a single-laser-shot intensity pattern from a large angular range (theta from 90 degrees to 168 degrees and phi from 0 degrees to 360 degrees) was detected from a single aerosol (e.g., a Bacillus subtilisspore, a 1-microm-diameter polystyrene latex sphere, or a cluster of either of these) flowing through the reflectors focal volume at 5 m/s. Noticeable difference in the large-angle-range two-dimensional angular optical scattering (LATAOS) suggest that the LATAOS pattern could be useful in differentiating and classifying life-threatening aerosols from normal background aerosols.

White-light nanosource with directional emission.

We report the first observation of white-light emission from femtosecond laser-induced plasma in a water droplet. Such emission is not observed with water in a cell. The microdroplet acts as a lens, focusing the incident light to nanosized regions within itself and directing the emission from these regions primarily back toward the laser source. This focusing increases the intensity so that multiphoton ionization generates plasma and causes it to reach the critical density during the initial part of the pulse, enabling the rest of the pulse to heat the plasma enough to emit in the visible.

Backward-enhanced fluorescence from clusters of microspheres and particles of tryptophan.

Measured fluorescence from single-particle clusters of dye-doped polystyrene microspheres, dried nonspherical particles of tryptophan, and single polystyrene microspheres is enhanced in the backward direction (180 degrees from the incident laser). This enhancement (a factor of 2-3 compared to 90 degrees), which can be interpreted as a consequence of the reciprocity principle, increases with the particle refractive index.