Vasanthi Sivaprakasam

Multiple UV wavelength excitation and fluorescence of bioaerosols

A two-wavelength excitation bioaerosol sensor has been developed and characterized for classifying various types of aerosols, including biological organisms and non-biological interferents. Single aerosols, smaller than 10 μm, are interrogated with 266 nm and 355 nm laser pulses separated in time by 400 ns. Fluorescence signals excited by these pulses are detected in three broad spectral bands centered at 350 nm, 450 nm and 550 nm. The results indicate that bacterial spores, vegetative bacterial cells and proteins can be differentiated based on the two wavelength excitation approach.

Spectral characterization of biological aerosol particles using two-wavelength excited laser-induced fluorescence and elastic scattering measurements

A two-wavelength laser-induced fluorescence (LIF) instrument has been developed and used to characterize individual biological aerosol particles, including biological warfare (BW) agent surrogates. Fluorescence in discrete spectral bands from widely different species, and also from similar species under different growth conditions were measured and compared. The two-wavelength excitation approach was found to increase discrimination among several biological materials, and especially with respect to diesel exhaust particles, a common interferent for LIF BW detection systems. The spectral characteristics of a variety of biological materials and ambient air components have been studied as a function of aerosol particle size and incident fluence.

Classification and selective collection of individual aerosol particles using laser-induced fluorescence.

We describe the development and performance evaluation of a system for optical interrogation, subsequent selection, and collection of individual aerosol particles entrained in an inlet air stream. Elastic scatter and laser-induced fluorescence obtained from single particles on-the-fly provide compositional information for classification criteria. Individual particles could then be selectively electrically charged and captured to a conductive substrate with an electric potential. The optical subsystem also includes a novel two-beam velocimeter to provide accurate downstream timing. Good overall quantitative performance values are reported for particles in the size range of 1-8 μm at mean rates up to 4 kHz.

Optical classification of bioaerosols using UV fluorescence and IR absorption spectroscopy

A partnership that includes the Naval Research Laboratory (NRL), MIT Lincoln Laboratories and the Edgewood Chemical and Biological Command is engaged in an effort to develop optical techniques for the rapid detection and classification of biological aerosols. This paper will describe two efforts at NRL: development of an improved UV fluorescence front-end trigger and the use of infrared absorption spectroscopy to classify biological aerosol particles. UV Laser-induced fluorescence (UVLIF) has been demonstrated to provide very high sensitivity for differentiating between biological and inorganic aerosol particles. Unfortunately, current UVLIF systems have unacceptably high false alarm rates due to interferences from man made and naturally occurring organic and biological particulates. We have developed a two-wavelength, UVLIF technique that offers a higher level of discrimination than is possible using single wavelength UVLIF. Infrared absorption spectroscopy coupled with multivariate analysis demonstrates a high potential for differentiation among members of biological and chemical sample classes. Two-wavelength UVLIF in combination with the IR interrogation of collected bioaerosols could provide a rapid, reagentless approach to specific classification of biological particles according to an operational level of discrimination - the degree of particle characterization required in order to signal the presence of pathogenic material.

Continuous flow, explosives vapor generator and sensor chamber.

A novel liquid injection vapor generator (LIVG) is demonstrated that is amenable to low vapor pressure explosives, 2,4,6-trinitrotoluene and hexahydro-1,3,5-trinitro-1,3,5-triazine. The LIVG operates in a continuous manner, providing a constant and stable vapor output over a period of days and whose concentration can be extended over as much as three orders of magnitude. In addition, a large test atmosphere chamber attached to the LIVG is described, which enables the generation of a stable test atmosphere with controllable humidity and temperature. The size of the chamber allows for the complete insertion of testing instruments or arrays of materials into a uniform test atmosphere, and various electrical feedthroughs, insertion ports, and sealed doors permit simple and effective access to the sample chamber and its vapor.

Field test results and ambient aerosol measurements using dual wavelength fluorescence excitation and elastic scatter for bioaerosols

A bioaerosol sensor based on dual wavelength fluorescence excitation and multiple wavelength elastic scattering has been developed and characterized for classifying micron-sized particles on the fly. The UVLIF instrument successfully completed a field trial in which we detected and correctly identified over 90% of the simulant releases over the 2 week testing period.

Single aerosol particle selection and capture using laser scattering and fluorescence

A dual wavelength UV-LIF fluorescence system that uses 266 nm and 355 nm laser pulses to sequentially excite single aerosol particles has been shown to provide significant discrimination between biological and ambient as well as differentiation among classes of biological particles. This particle classification data can then be used to trigger an electrostatic capture mechanism to deposit individual potential bio-threats particles onto a stainless steel substrate and particles that are not classified as targets are discharged with the exiting airflow. Timing and velocity information for each on-the-fly particle are critical for setting an appropriate delay to capture the particles of interest. A novel CW laser beam technique has been developed to measure the velocity of each particle and initiate a timing sequence. The electrostatic capture mechanism then electrically charges identified particles and produces a time-delayed electric field to drive them into the stainless steel substrate. The resulting collected sample is highly enriched with target, or potential threat, particles in comparison to their percentage in the ambient air. This presentation will describe the unique optical interrogation and diagnostic techniques that have been developed to make this achievement possible, as well as provide the latest system performance results.

Metallic-nanoparticles-enhanced fluorescence from individual micron-sized aerosol particles on-the-fly

Fluorescence spectra from individual aerosol particles that were either coated or embedded with metallic nanoparticles (MNPs) was acquired on-the-fly using 266 nm and 355 nm excitation. Using aqueous suspensions of MNPs with either polystyrene latex (PSL) spheres or dissolved proteins (tryptophan or ovalbumin), we generated PSL spheres coated with MNPs, or protein clusters embedded with MNPs as aerosols. Both enhanced and quenched fluorescence intensities were observed as a function of MNP concentration. Optimizing MNP material, size and spacing should yield enhanced sensitivity for specific aerosol materials that could be exploited to improve detection limits of single-particle, on-the-fly fluorescence or Raman based spectroscopic sensors.

Green and ultraviolet pulse generation with a compact, fiber laser, chirped-pulse amplification system for aerosol fluorescence measurements

We use a compact chirped-pulse amplified system to harmonically generate ultrashort pulses for aerosol fluorescence measurements. The seed laser is a compact, all-normal dispersion, mode-locked Yb-doped fiber laser with a 1050 nm center wavelength operating at 41 MHz. Average powers of more than 1.2 W at 525 nm and 350 mW at 262 nm are generated with <500 fs pulse durations. The pulses are time-stretched with high-dispersion fiber, amplified by a high-power, large-mode-area fiber amplifier, and recompressed using a chirped volume holographic Bragg grating. The resulting high-peak-power pulses allow for highly efficient harmonic generation. We also demonstrate for the first time to our knowledge, the use of a mode-locked ultraviolet source to excite individual biological particles and other calibration particles in an inlet air flow as they pass through an optical chamber. The repetition rate is ideal for biofluorescence measurements as it allows faster sampling rates as well as the higher peak powers as compared to previously demonstrated Q-switched systems while maintaining a pulse period that is longer than the typical fluorescence lifetimes. Thus, the fluorescence excitation can be considered to be quasicontinuous and requires no external synchronization and triggering.

Generation and optical characterization of aerosol particles with controlled mixed composition.

A method for controlled generation of composite aerosol particles is achieved by coating a core particle material, such as glass or polymer beads, with a second (analyte) material on the core surface. The mass fraction of the analyte can be varied over a wide range to generate resultant composite aerosol particles, which for the low end of analyte mass fractions has little influence on the particle size, but can be varied up to mass fractions nearly equivalent to the core material, as demonstrated in this paper. Analysis of this method was carried out using fluorescent analyte and core particle materials in separable spectral bands to measure both particle size distributions and fluorescent emission distributions on individual particle basis.