1. 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

2. 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

3. 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 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.

4. 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

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

6. 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

7. 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

8. What is “Instantaneous Microbial Detection”?
Can do:
Single particle detection, size and count
Resolution to +/ 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

9. 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.

10. 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
Wavelength (nm)
Wavelength (nm)

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

12. 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

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

14. 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

15. 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

16. 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

17. 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

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