Dr. Frank E. Inscore is a research and development professional with over 15 years of experience in chemistry, materials science, spectroscopy, and leadership of small start-up companies developing new technologies. His background includes a PhD in chemistry from the University of Arizona and postdoctoral research at the University of New Mexico, where he used techniques like X-ray absorption spectroscopy, magnetic circular dichroism, electronic absorption spectroscopy, and resonance Raman spectroscopy to study the structural and functional role of pyranopterin cofactors in molybdenum and tungsten enzymes. He has since worked in industrial research developing portable chemical analyzers and sensors using surface-enhanced Raman spectroscopy to identify chemicals and pathogens.

1. Dr. Frank E. Inscore
Research & Development Professional
“delivering excellence in leadership & management and success driving
innovation in new technology, application & product development”

2. OVERVIEW / OUTLINE
INTRODUCTION
Chemistry / Materials / Spectroscopy
Small Start-Up Company : Leadership / Management
KEY / 15+ YEARS EXPERIENCE / TRANSFERABLE
• I. PhD / Post-Doctoral Research Experience
• II. Industrial Research / Development Experience
CONCLUSION / FUTURE RELEVANCE
Q&A

3. I. PhD / Post-Doctoral Research Experience
Enemark Research Group
Dept. of Chemistry
University of Arizona
Kirk Research Group
Dept. of Chemistry
University of New Mexico
National Institutes of Health
National Science Foundation
Petroleum Research Fund
Sandia National Laboratories
Postdoc 2000 - 2003
Prof. John H. Enemark
Ph.D. 2000
Prof. Martin L. Kirk
National Science Foundation
Mo
S
S
O
Mo
S
S
S
Mo
S
S
O
S
SO
S-Cys
O
S er-O
OH2
Sulfite Oxidase Xanthine Oxidase DMSO Reductase
Pyranopterin Molybdenum and Tungsten Enzymes
HN
N N
N
OH2N OPO
3
2-
S-
S-
O H
H
Pyranopterin cofactor
• XAS / EXAFS [Mo(VI,V,IV)]
• MCD/ EPR [Mo(V)]
• Electronic Absorption
• Resonance Raman
Primary Issue
What is Structural and Functional Role of the
Pyranopterin Ene-1,2-Dithiolate Unit During
Course of Catalysis?
• Single Crystal XRD

4. Chemistry : Synthesis of Mo / W Dithiolene Systems
S
S
O
MN N
N N
N
N
S
S
MoN N
N N
N
N
S
S
S
MoN N
N N
N
N
N
O
HB
H3C
H3C
H3C
CH3
CH3
CH3
HB
H3C
H3C
H3C
CH3
CH3
CH3
HB
H3C
H3C
H3C
CH3
CH3
CH3
0,-1
M
S
S
M
S
SS
S
M
S
S
O
S
S
M
S
S
S
S
S
S
0,+1
-1,-2
-1,0
0,-1,-2
M
S
S
O
S
S
M
S
S
OR
S
S
M
S
S
O
S
S
M
Y
SR
O
S
S
M
Y
SR
O
S
S
RO
O
O
-1
-1
-2
-2
-1
Four Prototypical Ene-1,2-Dithiolate Systems:
S
S-S
-S
-S
-S
H
H N
N
-S
-S-S
-S- S
- S
M
S
S =

5. -4 10
-6
-3 10
-6
-2 10
-6
-1 10
-6
0
1 10
-6
2 10
-6
3 10
-6
4 10
-6
-1500.0-1000.0-500.000.0000500.001000.01500.0
(Tp*)MoO(bdt) (1)
I
II
1 5.7 14.7 13.7 12.7 11.7 10.7 9.7 8.7 7.7 6.7 5.7
Ionization Energy (eV)
C27
C23
C24
C17
C13
C14
C16
O
MO
C15
C26
N21
B
N32
C37
C33
C34
C36
N31
S1 C1 C6
C5
C4
C3
C2
S2
Resonance Raman Spectroscopy
Electrochemistry Magnetic Circular Dichroism
Electronic AbsorptionPhotoelectron Spectroscopy
Electron Paramagnetic Resonance
X-ray Crystallography
Density Functional Theory
Synthesis/ Purification
Characterization:
NMR, IR, HR-MS, XAS
Minimal
Structural/ Effective
Spectroscopic Active Site
Models
-1 10
5
-5 10
4
0
5 10
4
1 10
5
1.5 10
5
3100 3200 3300 3400 3500 3600 3700
Field (Gauss)
n= 9.4510 GHz
(3)
Physical Characterization: The (Tp*)MoO(bdt) Benchmark

12
3
5
67
4
a''op
a''ip
a'op
a'ip
a'z2
a''x2-y2
a''x'za'y'z
a'xy
O
O
M
= 90 0
M
a'
xy + a'
ip
S
S
> 90 0
a'
xy + a'
op a'
yz + a'
op

6. MCD and Electronic Absorption Spectroscopy
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
-1000
-500
0
500
1000
8000 12000 16000 20000 24000 28000 32000
Absorbance
MCDIntensity(mdeg)
Energy (wavenumbers)
1 2 (3, 4) 5 6 7
Complimentary selection rules:
resolve electronic transitions in spectra
MCD (5K /7T)
Low Temperature Solid-State (PDMSO Mull) Studies
Absorption (5K)

7. Band Assignments from Combined Spectroscopic Approach
0
800
1600
2400
3200
4000
4800
5600
6400
8000 12000 16000 20000 24000 28000 32000
Epsilon(M
-1
cm
-1
)
Energy (wavenumbers)
1 2 (3, 4)
5
6
7

1 2
3
5
6 7
4
a''
op
a''
ip
a'
op
a'
ip
a'
z2
a''
x2-y2
a''
x'z a'
y'z
a'
xy
O
O
M
= 90 0
M
a'
xy + a'
ip
S
S
> 90 0
a'
xy + a'
op a'
yz + a'
op
Solution Electronic Absorption (DCE)
Gas-phase PES / DFT

8. The Resonance Raman Experiment

9. Vibrational Analysis & Raman Excitation Profiles
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0
0.5
1
1.5
2
16000 18000 20000 22000 24000
Energy (wavenumbers)
S
S
M
O
3( A' )
S
S
M
O
S
S
M
O
1( A' )
6( A' )
(Tp*)MoO(bdt): 8K PDMSO mull EA; 100K RR NaCl/ Na2SO4
Observe large differential enhancement of Mo=O
Key Points from solid-state / solution studies
Transitions probed are orthogonal (in-plane vs out-of-plane)
Conclusions:
Sip  Mo dxy CT probes covalent contributions to ground-state
S Mo CT probes electronic contributions to redox potentials
Sip  Mo dxy
Sop  Mo dxz,yz
3 bands observed
polarized (A’ symmetry)

10. Implications for Catalytic Reactivity in Enzymes
xy
a'
ip
a'
xy
a'
ip
a'
S-Moxy
3-center
pseudo- antibonding
S-Moxy
3-center
pseudo- bonding
S
S
S
O
cys
Mo
OH2
(IV)
S
S
S
O
cys
Mo
OH
(V)
S
S
S
O
cys
Mo (VI)
O
SO3
2-
SO4
2-
H2O
H+
, e-
H+
, e-
Lowest energy (intense) CT must be Sip  Mo dxy
This CT transition probes covalency contributions to ET pathway.
Criteria for efficient ET
Good M-L overlap/ Minimize ROE
Reason Nature has chosen ene-1,2-dithiolate and M=O groups
M=O aligns redox orbital for facile ET via unique 3-center 2-electron bond.

11. Providing Chemical Information When & Where You Need It
Mission:
To provide superior chemical analyzers
(faster, portable, easy to use, rugged, more sensitive, less expensive)
To meet specific needs of
Department of Defense (Fuel Analysis, IED Identification)
Homeland Security (CWA, BWA, IED Identification)
Chemical Manufacturing Industry (Process Control)
Medical - Pharmaceutical (Drugs, HIV, TB)
Food - Beverage (food borne pathogens, pesticide contamination)
General Information:
Experience: 2003 - 2013 Director / Manager R&D (Raman-SERS Applications)
Products: RamanPro, RamanID, Portable Fuel Analyzer, and SERS-ID;
SERS Sensor vials, capillaries, microplates, disks, chips;
Sample probes, readers
Spectral analysis software / libraries
Services : Custom applications - analyzers, analysis, sensors / chemistries
11
II. Industrial Research Experience

12. RTA’s Raman Analyzers
Advantages:
• No sample preparation
• Simple integration via fiber
optics
• Remote analysis, multi-
component
• No fluorescence interference
• Complete spectral coverage
• Wavelength stability
• No analyzer calibration required
• Confident spectral subtraction
• and library search/match
• Real-time, On-demand analysis
• Long term stability
• Temperature and vibration
immune
• Operate from 25 to 100 oF
• Shock resistant
It’s an Interferometer…no x-axis drift!
Providing Chemical Information When & Where You Need It

13. RTA Raman Advantage: No Fluorescence
More Chemical Information
No Sample Preparation
785 nm
Laser
Excitation
1064 nm
Laser
Excitation
Diesel
Fuel
13
Providing Chemical Information When & Where You Need It
Virtually all natural materials fluoresce
using 785 nm
Wavenumbers (cm-1)
Raman
Infrared
Near Infrared

14. RTA Advantage: No X-Axis Shift
Aspirin
Aspirin Shifted
Ibuprofen
14
Providing Chemical Information When & Where You Need It
X axis shifts limit: quantitation, multi-component
analysis, and unknown identification

15. Raman Applications: Process Chemical Analyzer
In-Process, Continuous Analysis!
No Sample Preparation!
Providing Chemical Information When & Where You Need It
Fluorescence Free Raman
Analyzer Always Calibrated
Universal Interface
Industry Hardened
Biofuel Monitoring
NeSSi Applications
Microreactors
Drug Synthesis

16. Portable Fuel Analyzer
Fuel Properties in 3 minutes!
No Sample Preparation!
Providing Chemical Information When & Where You Need It
Cloud Pt
Pour Pt
Flash Pt
Freeze Pt
Distill Pts
Cetane
Density
Lubricity
Net Heat
Viscosity
Acid #
Aromatics
Olefins
Saturates
Sulfur
Water

17. Identify PPM chemicals in 10 seconds:
Field SERS ID Analyzer
Drugs
Explosives
Food Adulterants
Pathogens
Chemical Agents
Pesticides
Water Pollutants
Providing Chemical Information When & Where You Need It

18. ALSO, chemical contribution
can provide additional 103
enhancement
Sub part-per-billion detection
becomes possible with SERS
SERS
provides
Single Molecule Detection
some argue this requires
enhancement factor (EF)
of 1012 -1014
Others say 105 -108
Surface-enhanced Raman Spectroscopy
Raman, although weak effect,
provides molecular specificity
BUT, when a molecule is within
a laser induced plasmon field,
the efficiency of Raman scattering
increase’s by 106 i.e. 1 million times!
Sub part-per million detection possible
h
Plasmon Field

H
N
H
H
H
H
N
N
N
N
Ag
Surface-Enhanced
Raman Photon

19. Commercial SERS Substrates:
Benzenethiol
10-3M
10-5M
10-8M
(~10ppb)
benzenethiol
RTA: LMC ~10-11M (0.01 ng/mL or 10 ppt)
Conc. EF
102
104
107

20. Approach: RTA SERS Patented Sampling Systems
provide instant response in seconds as opposed to 30-min or more
Metal Particle
Sol-Gel Matrix
Adsorbed
Molecules
Molecules
in Solution
Laser
Raman
Scattering
Simple SERS Sample Vials
1 10
SER-Active Capillary
silver gold
SERS Microplate
High Throughput Screening Extraction and Pre-Concentration
RTA Patents
6,623,977; 6,943,031; 6,943,032; 7,312,088; 7,393,691; 7,393,692; 7,462,492; and 7,462,493
SERS spin-coated Disk

21. Functionalized Sol-Gel SERS
(affords greater selectivity and sensitivity)
PC OTC
Std chromatographic media
Patents: 6623977, 6943031, 6943032, 7312088,
7393691, 7393692, 7462492, 7462493, 7713914
R&D: SERS Lab-On-Chip

22. Example - The Overall Need/Problem
Detection of Bioagents
• Intentional Dispersal by Terrorist
• Domestic/Military Targets
• e.g. weaponized aerosols, contamination of water & food supplies
• B. anthracis, Y. pestis, F. tularensis, C. botulinum A, P. hanta
Detection of Waterborne Pathogens
• Unintentional Water Contamination
• Domestic/Military
• Cryptosporidium, Giardia, V. cholerae, Campylobacter jejuni
Detection of Foodborne Pathogens
• Unintentional Food Contamination
• Domestic/Military
• Listeria monocytogenes, E. coli (O157:H7), Salmonella enterica
Detection of Clinical Pathogens
• Unintentional Spread of Infections
• Domestic/Military Hospitals
• S. aureus (MERSA), Tuberculosis, AIDs
BA spores –almost perfect bioagent.
Most likely to be employed by terrorist :
right size so can be inhaled – most lethal route.
RTA is developing complete package to
address each of these application areas.

23. The Challenge & Goal
Detect bioagents and pathogens
on surfaces, in aerosols, water, in biofluids, and food.
(Category A bioagents 1st priority: Bacillus anthracis spores)
The device must provide the following:
• Sensitivity: Detect 10,000 spores Anthrax LD50 ~10,000 spores (100 ng)
• Speed: Within 15 minutes
• Specificity: Identify and discriminate pathogens
(No False Positives!)
• Reproducibility: Accurate and Repeatable
(No False Negatives!)
• Current methods:
• DNA or RNA enumeration: (Culture growth - 24 hours)
or Polymerase Chain Reactions (PCR, >4 hours)
• Test kits (limited shelf life, very high false positive rate)

24. Preliminary Analysis of Spores: Raman Spectroscopy
B. cereus
B. subtilis
B. anthracis
CaDPA
Core Wall
(proteins-cysteine,
)Ca dipicolinate
Cortex
(peptidoglycan)
Exosporium
Spore Coat
DNA
Ribosomes
C C
O O
O ON
2-
Ca 2+
J. Raman Spectrosc., 35, 82-86 (2004); Spectrosc., (2005)
Caveat: no consensus spectra in literature.
Variability: growth/media conditions.
Limitations: sensitivity/sample issues.
Portable 1064 nm system: Chem ID, Hazmat & Hoax Material Analyzer
PROBLEM: need both high Specificity & Sensitivity

25. The Solution: SERS
Inherent specificity: all chemicals/biochemicals produce a unique Raman
spectrum allowing unequivocal identification (no false-positives).
Improved sensitivity: Ag and Au nanoparticle substrates used to generate
SERS amplify Raman signals (increase scattering efficiency) by 1 million times
or more with potential detection at sub-ppb, i.e. 10-8 M (no false-negatives).
Enhancement Factor ~ 105
SERS 1ppm DPA (aq)
RS 20,000 ppm DPA (aq)
RS 83,000 ppm DPA (KOH)
RS solid DPA
100 mW, 1-min
290 mW, 5-min
100 mW, 5-min
290 mW, 5-min
DPA (dipicolinic acid)
LMC = ~ 1 ppb
1ppm DPA
83,000 ppm DPA
SERS
RS
FT-R 785 nm

26. APPROACH: Molecular Recognition Elements
(peptides / antibodies / aptamers)
Example - Peptide-Functionalized Silver Sol-gel
• Ag nanoparticles immobilized within porous sol-gel in a glass capillary were successfully functionalized
with a short peptide that specifically binds Bacillus anthracis spores.
• Peptide functionalization and spore binding are verified by SERS.
• The functionalized spectrum dominated by sulfur of the pep-cys-Ag bond (660 cm-1 ).
SERS of
BA specific peptide
cys-Ag
Cysteine-Ag
Peptide
Ag
Sol-Gel
Glass Capillary Wall
wash
pictures are not to scale
pep-cys-Ag
tf = 0.5-72 hrs
Use Immediately or Seal (stable >6-months)functionalize

27. ASSAY: Spore Capture - Incubation and Wash
Signal Amplification - SERS Enhancer Treatments
A 5-step SERS assay was successfully developed.
Peptide
Ag
Sol-Gel
Glass Capillary Wall
Spore
1) Draw 10 µL sample into a peptide functionalized capillary, wait 5-15 minutes.
2) Perform washes to minimize non-specific interactions (40 sec)
3) Treat sol-gel capture matrix with new proprietary reagent wash (10-sec)
4) Perform additional treatment using AA wash (10-sec)
sol-gel & spore treatmentscleaning washes
washes applied after spore incubation:
minimize non-specific interactions within capture matrix
while
treatments amplify spore signal via DPA enhancement
incubation

28. 5) Place capillary in Raman analyzer, measure spectrum (1 minute).
Spore Detection & Sensitivity: SERS
A 5-step SERS assay for BA was successfully developed.
(also similar BC and BS assays)
Sensitivity:
10 spore spectrum!
NO False Negatives!
1/100th of 1000 spores/mL sample is in capillary
measure
t = 60 seconds
Total Assay: 7-17 min
depends on spore
incubation time
(5-15 min)

29. Spore Assay Specificity / Repeatability
A 5-step SERS assay for BA was successfully developed.
1/100th of 10000 BA spores/mL sample is in capillary
Selectivity:
BA 100 spore assay challenged:
10-100X [higher] BC, BM, & BS.
NO False Positives!
BA-S BC
BM
BS
Repeatability:
100 BA spores - 12 for 12
Capillaries!
No False Negatives!

30. VALIDATION : at US ARMY ECBC
B. anthracis - Ames spores

31. CONCLUSION: Future Relevance
KEY / 15+ YEARS EXPERIENCE / TRANSFERABLE
WILL DELIVER EXCELLENCE and EXPERTISE
- Chemistry / Materials / Spectroscopy
- Leadership / Management
- Anti-Counterfeiting / Multilayered Defense
- Nanomaterials / Fluorescence