Particle Counting in Non-Cleanrooms Instantaneous Microbial Detection Presentation to National Air Filtration Association Particle Counting in Non-Cleanrooms Instantaneous Microbial Detection What’s In It For You JP Jiang, PhD, Chief Technology Officer Chuck Bolotin, VP, Sales 9-24-05

Agenda About BioVigilant Laboratories Particle Counting in Non-Cleanrooms How Microbial Contamination Testing is Done Now What is “Instantaneous Microbial Detection”? How IMD Differs from How It’s Done Now Industry Applications for IMD IAQ Consulting Military and Homeland Security Pharmaceutical Manufacturing Medical Device Manufacturing Personal Care Food Processing Healthcare “Sick Buildings” Joint Business Opportunities

About BioVigilant Laboratories Mission Statement Apply our scientific capabilities in the service of our customers’ needs. Headquarters in world famous “Optics Valley”, Tucson, Arizona Highly skilled researchers, scientists, engineers -- 12 patents issued, 7 pending Initial focus was homeland security Novel application of Mie Scattering Only company to combine simultaneous determination of inert or biological Currently applying technology to commercial markets Strong management team Experienced: average of 23+ years per person in successful tech development and deployment Multi-disciplinary: optical physics, software development, mechanical engineering, bio-sciences, medical, manufacturing, semiconductor, government.

Particle Counting in Non-Cleanrooms Q: What is the primary problem using a conventional particle counter in a non-cleanroom environment? A: The device is easily overwhelmed by the quantity of particles. BioVigilant’s detector was designed to work in a non-cleanroom environment. Not overwhelmed by quantity of particles Continuous monitoring design enables trending Applications: QA in filter manufacturing Part of air filtration system in buildings Monitor emissions in “dirty” environment

How Microbial Contamination Testing is Done Now Traditional Methods Plate culture of viable cells Antibody immunoassay “Rapid Methods” Plate scanning cytometer Flow cytometer PCR

What is “Instantaneous Microbial Detection”? Instantaneous Microbial Detection (or “IMD”) n. The process that provides immediate and continuous quantification of microbes present in a specified volume of air or liquid. <BioVigilant

Relevant Particle Size Range Color Guide = biological = inert = virus pollen bacteria mold bio-agent dust smog Virus 0.2mm 0.5mm 1mm 5mm 50mm Particle size

What is “Instantaneous Microbial Detection”? Can do: Single particle detection, size and count Resolution to +/- 0.25 micron; highly accurate sizing Simultaneously and in real time determine if each individual particle is inert or biological Broad range of microbial detection No need for reagent Cannot do: Detect viruses Identify genus

What is “Instantaneous Microbial Detection”? Scientific Principle Microbes (bacteria, fungi) contain metabolites, which emit fluorescence under UV illumination. Intrinsic fluorescence emission is used as the marker for microbes. Deterministic single particle sizing / fluorescence measurements provide ability to differentiate pollens and smoke from bacteria. Methodology eliminates the need of reagent dye labeling and achieves instantaneous detection.

Fluorescence Excitation Spectra   Metabolite Excitation Wavelength Peak / (Range) Tryptophan 270 nm (220 nm – 300 nm) NADH 340 nm (300 nm – 410 nm) Riboflavin 400 nm (320 nm – 420 nm) Relative Fluorescence Intensity (AFU) (J.K. Li, et al, Biotechnol. Prog. Vol. 7, 21, 1991) Relative Fluorescence Intensity (AFU) NAD(P)H Riboflavin 300 320 340 360 380 400 300 320 340 360 380 400 Wavelength (nm) Wavelength (nm)

Fluorescence Emission Spectra (Hill et al, Field Ana. Chem. & Tech, 3(4-5), 221,1999)

What is “Instantaneous Microbial Detection”? Instantaneous Microbial Detector Key features: High resolution particle counter Integration of BioVigilant’s patented single path technology Individual particle analysis 0.5 – 20 μm range (high resolution) Simultaneous fluorescence detection Single wavelength UV laser source Targeting major metabolites Technical background Leverage bio-defense sensor technology Advent of UV laser sources Model IMD-A

Continuous, Simultaneous & Instantaneous Microbial Detection Display of Microbe-free Air (Yellow bars are inert particles.) Microbe-free air data collection

Continuous, Simultaneous & Instantaneous Microbial Detection Display of Dispersed Baker’s Yeast (Saccharomyces Cerevisiae) (Yellow bars are inert particles. Red bars are microbe counts.) Yellow bars: inert particles Red bars: microbe counts

How IMD Differs from How It’s Done Now Instantaneous vs. 1 – 9 days Information for bio-terror attack is time sensitive Throw out less production Fewer planned halts in production Unplanned halts in production Easier to remediate and verify remediation Value as sales tool to demonstrate contamination and remediation Continuous monitoring and real-time feedback vs. episodic testing and time-delayed feedback Able to trend, spot anomalies / problems earlier Lowers chances of contamination

Industry Applications for IMD IAQ Consulting As a sales tool to show what work needs to be done As a sales tool to verify that remediation worked Military and Homeland Security Trigger device against bio-terror attack HVAC In the open Pharmaceutical Manufacturing FDA regulations Internal requirements Reputation Legal exposure Recalls Replace much of: settle plates, particle detectors, rapid methods devices Significant, quantifiable, understood, urgent need

Industry Applications for IMD Medical device manufacturing Similar requirements to pharmaceutical manufacturing Personal care (cosmetics) Significant internal requirements Food processing Limited but valuable in certain areas, no strict regulatory requirements Hospitals Hospital Acquired Illnesses Legal exposure “Sick Buildings” Commercial Government Schools

Joint Business Opportunities Internal use as sales and process tool Distribution Licensing ?