NANOTECHNOLOGY AND BIOSENSORS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0207104
• Project No.: ARZT-1377160-R22-130
• Multistate No.: NC-_old1194
• Project Start Date: Oct 1, 2011
• Project End Date: Sep 30, 2016

Project Director
Yoon, JE.

Recipient Organization
UNIVERSITY OF ARIZONA
888 N EUCLID AVE
TUCSON,AZ 85719-4824

Performing Department
Agri & Biosystems Engineering

Non Technical Summary
Development of sensing and processing technologies based on nanoscale phenomena becomes increasingly relevant for our society as we continue to advance our knowledge of biological phenomena related to food, agriculture, environment and energy. As we understand the molecular mechanisms that underlie the emergence and spread of pathogens and their consequent impact on our agricultural and food systems, it becomes evident that the technology to intervene and mitigate their effects on public health need to be correspondingly small, i.e. within the realm of nanotechnology. According to the FY 2010-2015 strategic plan published by the USDA, one of the strategic goals for this period is to ensure that all of America's children have access to safe, nutritious and balanced meals. Within this goal, specific objective 3 states that we must protect public health by ensuring that food is safe. Reduction in the number of foodborne illness will require a comprehensive "farm-to-fork" risk assessment, coupled with appropriate technological measures to screen, identify, and eliminate offending food pathogens.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
404	4099	2000	100%

Knowledge Area
404 - Instrumentation and Control Systems;

Subject Of Investigation
4099 - Microorganisms, general/other;

Field Of Science
2000 - Chemistry;

Keywords

biosensors

mems

lab on a chip

microfluidics

protein array

food safety

foodborne pathogens

Goals / Objectives
3. Develop biological interfaces and sensing systems incorporating microfabrication and nanotechnology. 4. Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems.

Project Methods
Objective 3 - Device design: Focus is on the design and fabrication of novel biosensor platforms using conventional state-of-the-art fabrication facilities. Specific activities include: i) Fabrication of multi-array channels for simultaneous detection in one sample application using electrochemical and optical signal measurement, iv) Development of a multi-component protein nanoarray (also described in the previous objective), v) System design and protocol development, viii) Use of computational simulation and modeling techniques for rapid development and evaluation of MEMS/nano-scale devices. Objective 4: Development of technologies for interrogating the impact of nanotechnology on living organisms: The uncertainty behind the environmental impact of nanotechnology arises from the following premise: nanoscale components of a system have the potential to interfere with biological processes that also happen to operate at the nanoscale. Conversely, in order to study the mechanisms of interaction between nanoscale systems with the biological world, we will need to develop advanced nanotechnology-based biosensing and monitoring systems.

Progress 10/01/11 to 09/30/16

Outputs
Target Audience:Academic research; food safety, veterinary diagnostics, medical diagnostics, tissue engineering and emerging biotech industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?YEAR 1 (10/01/2011-9/30/2012): OUTPUTS: 1. A new collaborative research contract was made to me, from QIA (Animal, Plant & Fisheries Quarantine & Inspection Agency, South Korea), about developing a handheld polymerase chain reaction (PCR) device for rapid veterinary diagnostics. I have visited QIA in May-June 2012 to formulate the collaborative research, and three researchers from QIA have also visited my lab in December 2012 to check our progress. Discussion is currently made to commercialize this technology. 2. The same technology is being applied to detect blood infections. Two awards have been made in December 2012 (both projects start in January 2013), one from AZ Furnace program and the other from Tech Launch Arizona, with the aim of commercializing this technology. A spin-off company, Fast PCR Diagnostics, LLC, has been created in December 2012. Negotiations are currently being made, including option agreement, exclusive practice right, conflict of interest, etc. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Bumsang Kim, Tae Hoon Oh (visiting scholars); David J. You, Tu San Park (post-docs); Jessica R. Gamboa, Pei-Shih Liang, Christopher F. Fronczek, Dustin K. Harshman, Scott V. Angus (graduate students); Katherine E. McCracken, Wenyue Li, Samir Mohandes, Roberto Reyes, Benjamin Erlick, Mariana Yanes, Lily Walsh, Katrina DeCook, Dalton Sycks (undergraduate students). (2) Partner organization: Animal, Plant & Fisheries Quarantine & Inspection Agency (QIA), South Korea. (3) Collaborators: Jae-Young Song (QIA), Marvin J. Slepian (University of Arizona College of Medicine). TARGET AUDIENCES: academic research, food safety, veterinary diagnostics, medical diagnostics, tissue engineering, emerging biotech industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period. YEAR 2 (10/01/2012-9/30/2013): 1. With the new collaborative research contract from QIA (Animal, Plant & Fisheries Quarantine & Inspection Agency, South Korea), a handheld polymerase chain reaction (PCR) device has been developed for rapid veterinary diagnostics. My research team (me and two of my graduate students) have visited QIA in June-July 2013 to deliver the alpha prototype and conducted experimental demonstrations. Discussion is currently being made to commercialize this technology. 2. The same technology has been applied to detect blood infections. Two awards have been made, which have started in January 2013, one from AZ Furnace program and the other from Tech Launch Arizona, with the aim of commercializing this technology. A spin-off company, Fast PCR Diagnostics, LLC, has been created, Negotiations are currently being pursued, including option agreement, exclusive practice right, conflict of interest, etc. YEAR 3 (10/01/2013-9/30/2014): Two post-docs, 9 graduate students and 9 undergraduate students were mentored by the PI during the project period. YEAR 4 (10/01/2014-9/30/2015): One post-doc, 7 graduate students, and 6 undergraduate students were mentored by the PI during the project period. YEAR 5 (10/01/2015-9/30/2016): One post-doc, 6 graduate students, and 9 undergraduate students were mentored by the PI during the project period. How have the results been disseminated to communities of interest?YEAR 2 (10/01/2012-9/30/2013): Journal paper publications, book publication, conference presentations, and seminar presentations. YEAR 3 (10/01/2013-9/30/2014): 1. Academic research: through disseminating our work in professional society's annual meetings, including MicroTAS 2013 (International Conference on Miniaturized Systems for Chemistry and Life Sceicnes), Biosensors 2014 (World Congress on Biosensors), IBE 2014 (Annual Meeting of Institute of Biological Engineering), SFB 2014 (Society for Biomaterials Annual Meeting and Exposition), IEEE-NEMS 2014 (IEEE International Conference on Nano/Micro Engineered and Molecular Systems), as well as numerous seminars in universities and national laboratories. 2. Industry: through a series of discussion and seminars with industry. YEAR 4 (10/01/2014-9/30/2015): 1. Academic research: through disseminating our work in professional society's annual meetings, including KSBB 2014 (Korean Society of Biotechnology and Bioengineering Fall Meeting and International Symposium), MicroTAS 2014 (International Conference on Miniaturized Systems for Chemistry and Life Sciences), IBE 2015 (Annual Meeting of Institute of Biological Engineering), In Vitro Biology 2015 (In Vitro Cellular and Developmental Biology Meeting), ASABE 2015 (American Society of Agricultural and Biological Engineers Annual International Meeting), as well as numerous seminars in universities and national laboratories. 2. Industry: through a series of discussion and seminars with industry. YEAR 5 (10/01/2015-9/30/2016): 1. Academic research: through disseminating our work in professional society's annual meetings, including IBE 2016 (Annual Meeting of Institute of Biological Engineering), as well as numerous seminars in universities and national laboratories. 2. Industry: through a series of discussion and seminars with industry. What do you plan to do during the next reporting period to accomplish the goals?YEAR 2 (10/01/2012-9/30/2013): 1. Through the sabbatical leave approved for fall 2013 semester, establish solid collaborations with the top-notch researchers outside the University of Arizona. 2. Participate in multi-institutional and multi-disciplinary collaborative proposals (including research center and training grants). 3. Continue to produce top-quality, high-impact peer-reviewed publications. YEAR 3 (10/01/2013-9/30/2014): 1. Wire-guided droplet PCR technology have been applied for veterinary diagnostics (influenza A), blood infection, and endocarditis, which is very fast and works with blood and tissue samples. A handheld prototype has recently been developed. Further miniaturization and implementation to solve real-world problems will be pursued, along with securing extramural grants. 2. Current efforts in OOC have been focused on stem cell differentiation and cancer study, while our effort is the first of its kind towards environmental toxicology. Specifically, 1) the use of our novel nanoensemble surface towards OOC, and 2) connecting two OOCs (liver and kidney) in series will make our work to be the first-ever demonstration. A series of optimizations will be pursued, along with journal article publications and extramural grants. 3. Our paper microfluidics and smartphone-based optical detection can be used by general public, not just the healthcare or environmental professionals, due to its small size, ease-of-use, and rapid assay time. Prototype development will be the main focus in the next reporting period. YEAR 4 (10/01/2014-9/30/2015): 1. Continue to publish top-quality, high-impact peer-reviewed publications. 2. Submit bigger research grants to NIH, NSF, and USDA based on the current collaborative grants. YEAR 5 (10/01/2015-9/30/2016): 1. Continue to publish top-quality, high-impact peer-reviewed publications. 2. Submit bigger research grants to NIH, NSF, and USDA based on the current collaborative grants.

Impacts
What was accomplished under these goals? YEAR 1 (10/01/2011-9/30/2012): 1. Wire-guided droplet PCR technology have been applied for veterinary diagnostics (influenza A) and blood infection, which is very fast and works with blood and tissue samples. A handheld prototype is currently being developed. A PCT application has been filed. 2. A novel method of creating ensemble nanotextured surfaces have been applied to not only endothelial cells, but also fibroblasts and smooth muscle cells. Controllable surfaces, using different types of polymer/nanofiber coatings and electrowetting technology, are also being developed towards creating better cardiovascular devices. 3. A simple paper microfluidics and smartphone-based optical detection is being considered for water quality monitoring (Cryptosporidium and endocrine-disrupting chemicals) and food safety application (Salmonella). YEAR 2 (10/01/2012-9/30/2013): 1. Wire-guided droplet PCR technology has been applied for veterinary diagnostics (influenza A) and blood infection, which is very fast and works with blood and tissue samples. A handheld prototype has been developed. 2. A novel method of creating ensemble nanotextured surfaces have been applied to not only endothelial cells, but also fibroblasts and smooth muscle cells. Controllable surfaces, using different types of polymer/nanofiber coatings and electrowetting technology, have also been developed towards creating better cardiovascular devices. 3. A simple paper microfluidics and smartphone-based optical detection is being considered for water quality monitoring (Cryptosporidium and endocrine-disrupting chemicals) and food safety application (Salmonella). YEAR 3 (10/01/2013-9/30/2014): 1. Yoon has actively participated AZ Furnace program from Arizona Commerce Authority and Proof of concept program from Tech Launch Arizona, to develop an extremely rapid and reprogrammable PCR machine that is based on wire-guided droplet PCR technology. The device is aimed to be used as point-of-care medical diagnostics specifically for bloodstream infection and heart valve endocarditis. 2. Yoon has recently started collaborative research with Terrence J. Monks (University of Arizona, College of Pharmacy), to study the behavior of human liver and kidney upon exposure to environmental toxins, using the miniaturized mock-up system called organ-on-a-chip (OOC). 3. Yoon has recently received two contracts from Seoul VioSys, one for developing a paper microfluidic device with smartphone identification for urinalysis (focused on diagnosing urinary tract infections, UTI, and sexually transmitted diseases - STDs, specifically gonorrhea, and the other for direct detection of particulate matter from HEPA filters (used in an air purifier). 4. Yoon has also started collaborative research with Kelly A. Reynolds (University of Arizona, College of Public Health), to develop a smartphone-based immunosensor strip towards multiplexed detection of water quality parameters, including E. coli, Cr(VI), Cl, caffeine, hardness, and pH. YEAR 4 (10/01/2014-9/30/2015): 1. Designed, fabricated, and tested a revolutionary biosensor that detect the presence of nucleic acid from complacted tissue samples in less than 5 min, referred to as DOTS qPCR. This work has been highlighted in numerous news magazines and websites. 2. Designed, fabricated, and tested a paper-based strip coupled with a smartphone-based sensor that detect the presence of pathogens from wastewater, food, urine, and blood. 3. Secured an NSF RAPID grant in collaboration with David Galbraith (Plant Sciences). Also secured an extension grant from SWEHSC in collaboration with Terrence Monks (Pharmacology and Toxicology), and another extension grant from WET in collaboration with Kelly Reynolds (Public Health). YEAR 5 (10/01/2015-9/30/2016): 1. The second edition of "Introduction to Biosensors," solely authored by Jeong-Yeol Yoon, was published by Springer. 2. Developedsmartphone-based handheld biosensors for water quality applications. 3. Developed a portable organ-on-a-chip (OOC) device that can be used for evaluating environmental toxicants as well as drugs. 4. Secured a new grant from BIO5 Institute. The SWEHSC and WET grants have been renewed.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Scott V. Angus, Soohee Cho, Dustin K. Harshman, Jae-Young Song and Jeong-Yeol Yoon, "A Portable, Shock-Proof, Surface-Heated Droplet PCR System for Escherichia coli Detection," Biosensors and Bioelectronics, 2015, 74: 360-368.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Christopher F. Fronczek and Jeong-Yeol Yoon, "Biosensors for Monitoring Airborne Pathogens," JALA - Journal of Laboratory Automation, 2015, 20(4): 390-410.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Pei-Shih Liang, Ariana M. Nicolini, Kimberly L. Ogden and Jeong-Yeol Yoon, "Use of Biosensors in Secondary Education Classrooms," Transactions of the ASABE, 2015, 58(2): 181-190.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ariana M. Nicolini, Christopher F. Fronczek and Jeong-Yeol Yoon, "Droplet-Based Immunoassay on a 'Sticky' Nanofibrous Surface for Multiplexed and Double Detection of Bacteria Using Smartphones," Biosensors and Bioelectronics, 2015, 67: 560-569.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Tu San Park and Jeong-Yeol Yoon, "Smartphone Detection of Escherichia coli from Field Water Samples on Paper Microfluidics," IEEE Sensors Journal, 2015, 15(3): 1902-1907.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Pei-Shih Liang, Tu San Park and Jeong-Yeol Yoon, "Rapid and Reagentless Detection of Microbial Contamination within Meat Utilizing a Smartphone-Based Biosensor," Scientific Reports, 2014, 4: 5953.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Seong-In Cho and Jeong-Yeol Yoon, "Paper Microfluidics for Red Wine Tasting," RSC Advances, 2014, 4(46): 24356-24362.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Hyuck-Jin Kwon, Christopher F. Fronczek, Scott V. Angus, Ariana M. Nicolini and Jeong-Yeol Yoon, "Rapid and Sensitive Detection of H1N1/2009 Virus from the Aerosol Samples with a Microfluidic Immunosensor," JALA - Journal of Laboratory Automation, 2014, 19(3): 322-331.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Christopher F. Fronczek, Tu San Park, Dustin K. Harshman, Ariana M. Nicolini and Jeong-Yeol Yoon, "Paper Microfluidic Extraction and Direct Smartphone-Based Identification of Pathogenic Nucleic Acid from Field and Clinical Samples," RSC Advances, 2014, 4(22): 11103-11110.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: C. Christopher Stemple, Scott V. Angus, Tu San Park and Jeong-Yeol Yoon, "Smartphone-Based Optofluidic Lab-on-a-Chip for Detecting Pathogens from Blood," JALA - Journal of Laboratory Automation, 2014, 19(1): 35-41.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Dustin K. Harshman, Roberto Reyes, Tu San Park, David J. You, Jae-Young Song and Jeong-Yeol Yoon, "Enhanced Nucleic Acid Amplification with Blood in Situ by Wire-Guided Droplet Manipulation (WDM)," Biosensors and Bioelectronics, 2014, 53: 167-174.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Tu San Park, Wenyue Li, Katherine E. McCracken and Jeong-Yeol Yoon, "Smartphone Quantifies Salmonella from Paper Microfluidics," Lab on a Chip, 2013, 13(24): 4832-4840.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Phat L. Tran, Jessica R. Gamboa, Katherine E. McCracken, Mark R. Riley, Marvin J. Slepian and Jeong-Yeol Yoon, "Nanowell-Trapped Charged Ligand-Bearing Nanoparticle Surfaces - A Novel Method of Enhancing Flow-Resistant Cell Adhesion," Advanced Healthcare Materials, 2013, 2(7): 1019-1027.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Katherine E. McCracken, Phat L. Tran, David J. You, Marvin J. Slepian and Jeong-Yeol Yoon, "Shear- vs. Nanotopography-Guided Control of Growth of Endothelial Cells on RGD-Nanoparticle-Nanowell Arrays," Journal of Biological Engineering, 2013, 7: 11.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Pei-Shih Liang and Jeong-Yeol Yoon, "Optofluidic Lab-on-a-chip Monitoring of Subsurface Bacterial Transport," Biological Engineering Transactions, 2013, 6(1): 17-28.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Jessica R. Gamboa, Samir Mohandes, Phat L. Tran, Marvin J. Slepian and Jeong-Yeol Yoon, "Linear Fibroblast Alignment on Sinusoidal Wave Micropatterns," Colloids and Surfaces B: Biointerfaces, 2013, 104: 318-325.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Christopher F. Fronczek, David J. You and Jeong-Yeol Yoon, "Single-Pipetting Microfluidic Assay Device for Rapid Detection of Salmonella from Poultry Package," Biosensors and Bioelectronics, 2013, 40(1): 342-349.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: David J. You, Tu San Park and Jeong-Yeol Yoon, "Cell-Phone-Based Measurement of TSH Using Mie Scatter Optimized Lateral Flow Assays," Biosensors and Bioelectronics, 2013, 40(1): 180-185.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Scott V. Angus, Hyuck-Jin Kwon and Jeong-Yeol Yoon, "Field-Deployable and Near-Real-Time Optical Microfluidic Biosensors for Single-Oocyst-Level Detection of Cryptosporidium parvum from Field Water Samples," Journal of Environmental Monitoring, 2012, 14(12): 3295-3304.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: David J. You and Jeong-Yeol Yoon, "Droplet Centrifugation, Droplet DNA Extraction, and Rapid Droplet Thermocycling for Simpler and Faster PCR Assay Using Wire-Guided Manipulations," Journal of Biological Engineering, 2012, 6: 15.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Jeong-Yeol Yoon and Bumsang Kim, "Lab-on-a-Chip Pathogen Sensor for Food Safety," Sensors, 2012, 12(8): 10713-10741.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: C. Christopher Stemple, Hyuck-Jin Kwon and Jeong-Yeol Yoon, "Rapid and Sensitive Detection of Malaria Antigen in Human Blood with Lab-on-a-Chip," IEEE Sensors Journal, 2012, 12(9): 2735-2736.
• Type: Journal Articles Status: Published Year Published: 2011 Citation: Jae-Young Song, Chang-Hee Lee, Eun-Jin Choi, Keesung Kim and Jeong-Yeol Yoon, "Sensitive Mie Scattering Immunoagglutination Assay of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) from Lung Tissue Samples in a Microfluidic Chip," Journal of Virological Methods, 2011, 178(1-2): 31-38.
• Type: Journal Articles Status: Published Year Published: 2011 Citation: David J. You, Kenneth J. Geshell and Jeong-Yeol Yoon, "Direct and Sensitive Detection of Foodborne Pathogens within Fresh Produce Samples Using a Field-Deployable Handheld Device," Biosensors and Bioelectronics, 2011, 28(1): 399-406.
• Type: Books Status: Published Year Published: 2016 Citation: Jeong-Yeol Yoon, "Introduction to Biosensors: From Electric Circuits to Immunosensors," Second Edition, Springer: New York, 2016, ISBN: 978-3-319-27411-9.
• Type: Books Status: Published Year Published: 2013 Citation: Jeong-Yeol Yoon, "Introduction to Biosensors: From Electric Circuits to Immunosensors," Springer: New York, 2013. ISBN: 978-1-4419-6021-4.
• Type: Book Chapters Status: Published Year Published: 2016 Citation: Pei-Shih Liang, Tu San Park and Jeong-Yeol Yoon, "Light Scattering Based Detection of Food Pathogens," in Light Scattering Technology for Food Property, Quality and Safety Assessment, Editor: Renfu Lu, CRC Press (Taylor & Francis): Boca Raton, 2016, pp.429-444.
• Type: Book Chapters Status: Published Year Published: 2016 Citation: Dustin K. Harshman and Jeong-Yeol Yoon, "Wire-Guided Droplet Manipulation for Molecular Biology," in Microfluidic Methods for Molecular Biology, Editors: Chang Lu and Scott Verbridge, Springer: Switzerland, 2016, pp.235-252.
• Type: Book Chapters Status: Published Year Published: 2016 Citation: Christopher F. Fronczek and Jeong-Yeol Yoon, "Detection of Foodborne Pathogens Using Biosensors," in Antimicrobial Food Packaging, Editor: Jorge Barros-Velazquez, Academic Press (Elsevier): London/San Diego/Waltham/Oxford, 2016, pp.153-166.
• Type: Other Status: Published Year Published: 2014 Citation: Jeong-Yeol Yoon, "Smartphone-Based Lab-on-a-Chip Sensor for Flu Detection," Resource, 2014, 21(1): 20-22.
• Type: Other Status: Published Year Published: 2012 Citation: Jeong-Yeol Yoon, "Who We Are & What We Can Do," Resource, 2012, 19(3): 19-21.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Ariana M. Nicolini, Dustin K. Harshman, Tyler D. Toth, M. Alejandra Mandel, David W. Galbraith and Jeong-Yeol Yoon, "DOTS qPCR: A Handheld, Rapid Molecular Diagnostic Tool for Ebola," Annual Meeting of IBE, Greenville, SC, April 7-9, 2016.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Soohee Cho, Argel Islas-Robles, Ariana M. Nicolini, Terrence J. Monks and Jeong-Yeol Yoon, "In Situ Monitoring of Induced Nephrotoxicity in Organ-on-a-Chip with Smartphone-Based Fluorescence Microscope," Annual Meeting of IBE, Greenville, SC, April 7-9, 2016.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Robin E. Sweeney and Jeong-Yeol Yoon, "Rapid and Non-Destructive Detection of Tissue Bacterial Infection," Annual Meeting of IBE, Greenville, SC, April 7-9, 2016.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Soohee Cho, Tu San Park, Tigran G. Nahapetian and Jeong-Yeol Yoon, "Smartphone-Based, Sensitive uPAD Detection of Urinary Tract Infection and Gonorrhea," Annual Meeting of IBE, Greenville, SC, April 7-9, 2016.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Innovative qPCR using interfacial effects to enable low threshold cycle detection and inhibition relief," ASABE Annual International Meeting, New Orleans, LA, July 26-29, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Mobile paper-based water quality monitoring via smartphones," ASABE Annual International Meeting, New Orleans, LA, July 26-29, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Smartphone biosensors and organ-on-a-chip," In In Vitro Cellular & Developmental Biology, Tucson, AZ, May 30 - June 3, 2015, 51: S6S7.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "IBE today: a platform for the future - niche areas for IBE," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Biosensors for monitoring airborne pathogens," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Dustin K. Harshman, Brianna M. Rao, Jean E. McLain, George S. Watts and Jeong-Yeol Yoon, "Interfacial effects revolutionize qPCR by low threshold cycle detection and inhibition relief," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Ariana M. Nicolini, Soohee Cho and Jeong-Yeol Yoon, "Pro-adhesive extracellular matrix mimic for use on organ-on-achip," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Katherine E. McCracken, Scott V. Angus, Tu San Park, Kelley A. Reynolds and Jeong-Yeol Yoon, "Smartphone for water quality," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Jeong-Yeol Yoon, "Smartphone biosensors and organ-on-a-chip," 2014 KSBB Fall Meeting and International Symposium, October 5-7, 2014, Changwon, South Korea.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini and Jeong-Yeol Yoon, "Pro-Adhesive Extracellular Matrix Mimic for Use in Organ-on-a-Chip," The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014), San Antonio, TX, October 26-30, 2014, pp.760-762.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Dustin K. Harshman, Roberto Reyes and Jeong-Yeol Yoon, "Rapid Molecular Diagnosis of Infectious Endocarditis: Developing uREx Dx," The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014), San Antonio, TX, October 26-30, 2014, pp.1051-1053.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Scott V. Angus, Soohee Cho, Dustin K. Harshman and Jeong-Yeol Yoon, "Quantitative, Surface Heated, Droplet Polymerase Chain Reaction for Detecting Pathogens," The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014), San Antonio, TX, October 26-30, 2014, pp.1452-1454.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Cayla Baynes, Tu San Park, Jeong-Yeol Yoon, "Enhanced fluorescent light scatter detection of cancer biomarkers using paper microfluidics," Biosensors 2014: World Congress on Biosensors, Melbourne, Australia, 27-30 May 2014. Paper number O76.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini, Christopher F. Fronczek, Jeong-Yeol Yoon, "Rapid, single-step, droplet-based bacterial assay platform on a nanofibrous substrate," Biosensors 2014: World Congress on Biosensors, Melbourne, Australia, 27-30 May 2014. Paper number O112.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Jeong-Yeol Yoon, "Paper microfluidics for red wine tasting," Biosensors 2014: World Congress on Biosensors, Melbourne, Australia, 27-30 May 2014. Paper number P3.198.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Dustin K. Harshman, Roberto Reyes, Tu San Park, Jeong-Yeol Yoon, "Extremely fast nucleic acid amplification by droplet manipulation for point-of-care diagnosis of blood infection," Annual Meeting of IBE, Raleigh, NC, 7-9 March 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Phat L. Tran, Daniel A. Martin, Jessica R. Gamboa, Jeong-Yeol Yoon, Marvin J. Slepian, "Nanopost fence: a novel strategy of preventing smooth muscle cells topographic migration," Society For Biomaterials (SFB) 2014 Annual Meeting and Exposition, Denver, CO, 16-19 April 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Seong-In Cho, Jeong-Yeol Yoon, "Paper microfluidics for red wine tasting," The 9th IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Waikiki Beach, HI, 13-16 April 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini, Celine M. Cohn, Jessica R. Gamboa, Marvin J. Slepian, Xiaoyi Wu, Jeong-Yeol Yoon, "Fabrication of a pro-adhesive surface using electrospun PCL nanofibers interspersed with peptide conjugated polystyrene particles," The 9th IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Waikiki Beach, HI, 13-16 April 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini, Celine M. Cohn, Marvin J. Slepian, Xiaoyi Wu, Jeong-Yeol Yoon, "Fabrication of a pro-adhesive surface using electrospun PCL nanofibers interspersed with peptide conjugated polystyrene particles," Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Jeong-Yeol Yoon, "Smartphone-based paper microfluidic detection of E. coli from field or waste water," Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Scott V. Angus, Soohee Cho, Dustin K. Harshman, Jeong-Yeol Yoon, "Wire-guided droplet manipulation based quantitative PCR device towards food and veterinary diagnostics," Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Jeong-Yeol Yoon, "Paper microfluidics for red wine tasting," Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Tu San Park, Dustin K. Harshman, Christopher F. Fronczek and Jeong-Yeol Yoon, "Smartphone Detection of Escherichia coli from Wastewater Utilizing Paper Microfluidics," The 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2013), Freiburg, Germany, 27-31 October 2013, pp. 1347-1349.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Christopher F. Fronczek, Tu San Park and Jeong-Yeol Yoon, "Paper Microfluidic Extraction of Bacterial and Viral Nucleic Acid from Field and Clinical Samples towards a Direct MicroTAS Apparatus," The 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2013), Freiburg, Germany, 27-31 October 2013, pp. 1114-1116.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Dustin K. Harshman, Roberto Reyes, Jeong-Yeol Yoon, "Direct Detection of Plasmid-Mediated Antibiotic Resistance in Bloodstream Infection by PCR Using Wire-Guided Droplet Manipulation (WDM)," The 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2013), Freiburg, Germany, 27-31 October 2013, pp. 470-472
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Jeong-Yeol Yoon, Tu San Park, Wenyue Li, Pei-Shih Liang, "Paper microfluidics detection of Salmonella using a smartphone," ASABE Annual International Meeting, Kansas City, MO, 21-24 July, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Pei-Shih Liang, Jeong-Yeol Yoon, "Rapid detection of foodborne pathogens within meat utilizing a smartphone biosensor," ASABE Annual International Meeting, Kansas City, MO, 21-24 July, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Pei-Shih Liang, Marisa Kleiman, Jeong-Yeol Yoon, "Use of biosensor in secondary education curriculum to improve students interest and awareness of science, engineering, and current worldwide issues," ASABE Annual International Meeting, Kansas City, MO, 21-24 July, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Tu San Park, Wenyue Li, Jeong-Yeol Yoon, "Paper microfluidics detection of Salmonella using a smart phone," Annual Meeting of IBE, Raleigh, NC, 7-9 March 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: C. Christopher Stemple, Hyuck-Jin Kwon, Jeong-Yeol Yoon, "Rapid and Sensitive Detection of Malaria Antigen in Whole Blood Using a Handheld LOC Device," Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: David J. You, Phat L. Tran, Hyuck-Jin Kwon, Jeong-Yeol Yoon, "Extremely Rapid and Fully Reprogrammable Total PCR Assays Using Wire-Guided Droplet Microfluidics," Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: David J. You, Joel R. Ehrenkranz, Jeong-Yeol Yoon, "Cell-Phone-Based Measurement of TSH using Mie Scatter Optimized Lateral Flow Assays," Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Christopher F. Fronczek, David J. You, Jeong-Yeol Yoon, "Single-Pipetting Microfluidic Assay Device for Salmonella in Poultry Package Water," Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Katherine E. McCracken, Phat L. Tran, Marvin J. Slepian, Jeong-Yeol Yoon, "Nanoscale Patterning Under Shear Stress for Guided Endothelial Cell Growth," 2012 Annual Meeting of IBE, Indianapolis, IN, 1-3 March 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Jessica R. Gamboa, Phat L. Tran, Jeong-Yeol Yoon, "Linear Fibroblast Alignment on Sinusoidal Wave Micropatterns," 2012 Annual Meeting of IBE, Indianapolis, IN, 1-3 March 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Jessica R. Gamboa, Jeong-Yeol Yoon, Richard G. Smith, Marvin J. Slepian, "Evaluation of the Performance of a Left Ventricular Assist Device in a Novel In Vitro Heart Failure Model," 2012 Annual Meeting of IBE, Indianapolis, IN, 1-3 March 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Scott V. Angus, Hyuck-Jin Kwon, Jeong-Yeol Yoon, "Low-Level Detection of Cryptosporidium parvum in Field Water Using Optical Microfluidic Biosensors," Proceedings of SPIE, 2012, 8229: 82290F.
• Type: Conference Papers and Presentations Status: Published Year Published: 2011 Citation: Phat L. Tran, Jessica R. Gamboa, Katherine E. McCracken, Mark R. Riley, Marvin J. Slepian, Jeong-Yeol Yoon, "Nanowell-trapped Charged Ligand-bearing Nanoparticle Surfaces - a Novel Method for Enhancing Flow Resistant Cell Adhesion," BioInterface Workshop & Symposium, Minneapolis, MN, 24-26 October 2011.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Dustin K. Harshmann, Roberto Reyes, David J. You, Jeong-Yeol Yoon, "Device for Near-Instant Diagnosis of Clinical Infection by Convective Droplet Thermocycling and 16s rRNA Hybervariable Region Probes," BMES 2012 Annual Meeting, Atlanta, GA, 24-27 October 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: David J. You, Phat L. Tran, Hyuck-Jin Kwon, Jeong-Yeol Yoon, "Extremely Rapid and Fully Reprogrammable Total PCR Assays Using Wire-Guided Droplet Microfluidics," 2012 ASABE Annual International Meeting, Dallas, TX, 29 July - 1 August 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Christopher F. Fronczek, David J. You, Jeong-Yeol Yoon, "Single-Pipetting Microfluidic Assay Device for Salmonella in Poultry Package Water," 2012 ASABE Annual International Meeting, Dallas, TX, 29 July - 1 August 2012.
• Type: Conference Papers and Presentations Status: Published Year Published: 2011 Citation: Hyuck-Jin Kwon, Scott V. Angus, David J. You, C. Christopher Stemple, Jeong-Yeol Yoon, "Development of a Handheld Optofluidic Immunosensor to Track the Transport and Distribution of H1N1/2009 Virus in a Mock Classroom," The 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences (uTAS 2011), Seattle, WA, 2-6 October 2011, pp. 1421-1423.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Katherine E. McCracken and Jeong-Yeol Yoon, "Recent Approaches for Optical Smartphone Sensing in Resource-Limited Settings: A Brief Review," Analytical Methods, 2016, 8: 6591-6601.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Soohee Cho, Argel Islas-Robles, Ariana M. Nicolini, Terrence J. Monks and Jeong-Yeol Yoon, "In Situ, Dual-Mode Monitoring of Organ-on-a-Chip with Smartphone-Based Fluorescence Microscope," Biosensors and Bioelectronics, 2016, 86: 697-705.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Ariana M. Nicolini, Tyler D. Toth and Jeong-Yeol Yoon, "Tuneable Nanoparticle-Nanofiber Composite Substrate for Improved Cellular Adhesion," Colloids and Surfaces B: Biointerfaces, 2016, 145: 830-838.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Katherine E. McCracken, Scott V. Angus, Kelly A. Reynolds and Jeong-Yeol Yoon, "Multimodal Imaging and Lighting Bias Correction for Improved uPAD-based Water Quality Monitoring via Smartphones," Scientific Reports, 2016, 6: 27529.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Dustin K. Harshman, Brianna M. Rao, Jean E. McClain, George S. Watts and Jeong-Yeol Yoon, "Innovative qPCR Using Interfacial Effects to Enable Low Threshold Cycle Detection and Inhibition Relief," Science Advances, 2015, 1(8): e1400061.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Soohee Cho, Tu San Park, Tigran G. Nahapetian and Jeong-Yeol Yoon, "Smartphone-Based, Sensitive uPAD Detection of Urinary Tract Infection and Gonorrhea," Biosensors and Bioelectronics, 2015, 74: 601-611.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Academic research; food safety, veterinary diagnostics, medical diagnostics, tissue engineering and emerging biotech industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One post-doc, 7graduate students, and 6 undergraduate students were mentored by the PI during the project period. How have the results been disseminated to communities of interest?1. Academic research: through disseminating our work in professional society's annual meetings, including KSBB 2014 (Korean Society of Biotechnology and Bioengineering Fall Meeting and International Symposium), MicroTAS 2014 (International Conference on Miniaturized Systems for Chemistry and Life Sciences), IBE 2015 (Annual Meeting of Institute of Biological Engineering), In Vitro Biology 2015 (In Vitro Cellular and Developmental Biology Meeting), ASABE 2015 (American Society of Agricultural and Biological Engineers Annual International Meeting), as well as numerous seminars in universities and national laboratories. 2. Industry: through a series of discussion and seminars with industry. What do you plan to do during the next reporting period to accomplish the goals?1. Continue to publish top-quality, high-impact peer-reviewed publications. 2. Submit bigger research grants to NIH, NSF, and USDA based on the current collaborative grants.

Impacts
What was accomplished under these goals? 1. Designed, fabricated, and tested a revolutionary biosensor that detect the presence of nucleic acid from complacted tissue samples in less than 5 min, referred to as DOTS qPCR. This work has been highlighted in numerous news magazines and websites. 2. Designed, fabricated, and tested a paper-based strip coupled with a smartphone-based sensor that detect the presence of pathogens from wastewater, food, urine, andblood. 3. Secured an NSF RAPID grant in collaboration with David Galbraith (Plant Sciences). Also secured an extension grant from SWEHSC in collaboration with Terrence Monks (Pharmacology and Toxicology), and another extension grant from WET in collaboration with Kelly Reynolds (Public Health).

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ariana M. Nicolini, Christopher F. Fronczek and Jeong-Yeol Yoon, "Droplet-Based Immunoassay on a 'Sticky' Nanofibrous Surface for Multiplexed and Double Detection of Bacteria Using Smartphones," Biosensors and Bioelectronics, 2015, 67: 560-569.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Pei-Shih Liang, Ariana M. Nicolini, Kimberly L. Ogden and Jeong-Yeol Yoon, "Use of Biosensors in Secondary Education Classrooms," Transactions of the ASABE, 2015, 58(2): 181-190.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Christopher F. Fronczek and Jeong-Yeol Yoon, "Biosensors for Monitoring Airborne Pathogens," JALA - Journal of Laboratory Automation, 2015, 20(4): 309-410.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Scott V. Angus, Soohee Cho, Dustin K. Harshman, Jae-Young Song and Jeong-Yeol Yoon, "A Portable, Shock-Proof, Surface-Heated Droplet PCR System for Escherichia coli Detection," Biosensors and Bioelectronics, 2015, 74: 360-368.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Soohee Cho, Tu San Park, Tigran G. Nahapetian and Jeong-Yeol Yoon, "Smartphone-Based, Sensitive �PAD Detection of Urinary Tract Infection and Gonorrhea," Biosensors and Bioelectronics, 2015, 74: 601-611.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Dustin K. Harshman, Brianna M. Rao, Jean E. McClain, George S. Watts and Jeong-Yeol Yoon, "Innovative qPCR Using Interfacial Effects to Enable Low Threshold Cycle Detection and Inhibition Relief," Science Advances, 2015, 1(8): e1400061.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini and Jeong-Yeol Yoon, "Pro-Adhesive Extracellular Matrix Mimic for Use in Organ-on-a-Chip," The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014), San Antonio, TX, October 26-30, 2014, pp.760-762.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Dustin K. Harshman, Roberto Reyes and Jeong-Yeol Yoon, "Rapid Molecular Diagnosis of Infectious Endocarditis: Developing uREx Dx," The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014), San Antonio, TX, October 26-30, 2014, pp.1051-1053.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Scott V. Angus, Soohee Cho, Dustin K. Harshman and Jeong-Yeol Yoon, "Quantitative, Surface Heated, Droplet Polymerase Chain Reaction for Detecting Pathogens," The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014), San Antonio, TX, October 26-30, 2014, pp.1452-1454.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Katherine E. McCracken, Scott V. Angus, Tu San Park, Kelley A. Reynolds and Jeong-Yeol Yoon, "Smartphone for water quality," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Ariana M. Nicolini, Soohee Cho and Jeong-Yeol Yoon, "Pro-adhesive extracellular matrix mimic for use on organ-on-a-chip," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Dustin K. Harshman, Brianna M. Rao, Jean E. McLain, George S. Watts and Jeong-Yeol Yoon, "Interfacial effects revolutionize qPCR by low threshold cycle detection and inhibition relief," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Biosensors for monitoring airborne pathogens," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "IBE today: a platform for the future - niche areas for IBE," Annual Meeting of IBE, St. Louis, MO, March 5-7, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Smartphone biosensors and organ-on-a-chip," In In Vitro Cellular & Developmental Biology, Tucson, AZ, May 30 - June 3, 2015, 51: S6S7.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Innovative qPCR using interfacial effects to enable low threshold cycle detection and inhibition relief," ASABE Annual International Meeting, New Orleans, LA, July 26-29, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Jeong-Yeol Yoon, "Mobile paper-based water quality monitoring via smartphones," ASABE Annual International Meeting, New Orleans, LA, July 26-29, 2015.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Jeong-Yeol Yoon, "Smartphone biosensors and organ-on-a-chip," 2014 KSBB Fall Meeting and International Symposium, October 5-7, 2014, Changwon, South Korea.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Tu San Park and Jeong-Yeol Yoon, "Smartphone Detection of Escherichia coli from Field Water Samples on Paper Microfluidics," IEEE Sensors Journal, 2015, 15(3): 1902-1907.

Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Academic research; food safety, veterinary diagnostics, medical diagnostics, tissue engineering and emerging biotech industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Two post-docs, 9 graduate students and 9 undergraduate students were mentored by the PI during the project period. How have the results been disseminated to communities of interest? 1. Academic research: through disseminating our work in professional society's annual meetings, including MicroTAS 2013 (International Conference on Miniaturized Systems for Chemistry and Life Sciences), Biosensors 2014 (World Congress onBiosensors), IBE 2014 (Annual Meeting of Institute of Biological Engineering), SFB 2014 (Society for Biomaterials Annual Meeting and Exposition), IEEE-NEMS 2014 (IEEE International Conference on Nano/Micro Engineered and Molecular Systems), as well as numerous seminars in universities and national laboratories. 2. Industry: through a series of discussion and seminars with industry. What do you plan to do during the next reporting period to accomplish the goals? Wire-guided droplet PCR technology have been applied for veterinary diagnostics (influenza A), blood infection, and endocarditis, which is very fast and works with blood and tissue samples. A handheld prototype has recently been developed. Further miniaturization and implementation to solve real-world problems will be pursued, along with securing extramural grants. Current efforts in OOC have been focused on stem cell differentiation and cancer study, while our effort is the first of its kind towards environmental toxicology. Specifically, 1) the use of our novel nanoensemble surface towards OOC, and 2) connecting two OOCs (liver and kidney) in series will make our work to be the first-ever demonstration. A series of optimizations will be pursued, along with journal article publications and extramural grants. Our paper microfluidics and smartphone-based optical detection can be used by general public, not just the healthcare or environmental professionals, due to its small size, ease-of-use, and rapid assay time. Prototype development will be the main focus in the next reporting period.

Impacts
What was accomplished under these goals? Yoon has actively participated AZ Furnace program from Arizona Commerce Authority and Proof of concept program from Tech Launch Arizona, to develop an extremely rapid and reprogrammable PCR machine that is based on wire-guided droplet PCR technology. The device is aimed to be used as point-of-care medical diagnostics specifically for bloodstream infection and heart valve endocarditis. Yoon has recently started collaborative research with Terrence J. Monks (University of Arizona, College of Pharmacy), to study the behavior of human liver and kidney upon exposure to environmental toxins, using the miniaturized mock-up system called organ-on-a-chip (OOC). Yoon has recently received two contracts from Seoul VioSys, one for developing a paper microfluidic device with smartphone identification for urinalysis (focused on diagnosing urinary tract infections, UTI, and sexually transmitted diseases - STDs, specifically gonorrhea, and the other for direct detection of particulate matter from HEPA filters (used in an air purifier). Yoon has also started collaborative research with Kelly A. Reynolds (University of Arizona, College of Public Health), to develop a smartphone-based immunosensor strip towards multiplexed detection of water quality parameters, including E. coli, Cr(VI), Cl, caffeine, hardness, and pH.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Jeong-Yeol Yoon, Smartphone-based paper microfluidic detection of E. coli from field or waste water, Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Scott V. Angus, Soohee Cho, Dustin K. Harshman, Jeong-Yeol Yoon, Wire-guided droplet manipulation based quantitative PCR device towards food and veterinary diagnostics, Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Jeong-Yeol Yoon, Paper microfluidics for red wine tasting, Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.
• Type: Journal Articles Status: Awaiting Publication Year Published: 2014 Citation: Ariana M. Nicolini, Christopher F. Fronczek and Jeong-Yeol Yoon, "Droplet-Based Immunoassay on a Sticky Nanofibrous Surface for Multiplexed and Double Detection of Bacteria Using Smartphones," Biosensors and Bioelectronics, 2014, doi:10.1016/j.bios.2014.09.040.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Pei-Shih Liang, Tu San Park and Jeong-Yeol Yoon, "Rapid and Reagentless Detection of Microbial Contamination within Meat Utilizing a Smartphone-Based Biosensor," Scientific Reports, 2014, 4: 5953.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Seong-In Cho and Jeong-Yeol Yoon, "Paper Microfluidics for Red Wine Tasting," RSC Advances, 2014, 4(46): 24356-24362.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Hyuck-Jin Kwon, Christopher F. Fronczek, Scott V. Angus, Ariana M. Nicolini and Jeong-Yeol Yoon, "Rapid and Sensitive Detection of H1N1/2009 Virus from the Aerosol Samples with a Microfluidic Immunosensor," Journal of Laboratory Automation, 2014, 19(3): 322-331.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Christopher F. Fronczek, Tu San Park, Dustin K. Harshman, Ariana M. Nicolini and Jeong-Yeol Yoon, "Paper Microfluidic Extraction and Direct Smartphone-Based Identification of Pathogenic Nucleic Acid from Field and Clinical Samples," RSC Advances, 2014, 4(22): 11103-11110.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: C. Christopher Stemple, Scott V. Angus, Tu San Park and Jeong-Yeol Yoon, "Smartphone-Based Optofluidic Lab-on-a-Chip for Detecting Pathogens from Blood," Journal of Laboratory Automation, 2014, 19(1): 35-41.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Dustin K. Harshman, Roberto Reyes, Tu San Park, David J. You, Jae-Young Song and Jeong-Yeol Yoon, Enhanced Nucleic Acid Amplification with Blood in Situ by Wire-Guided Droplet Manipulation (WDM), Biosensors and Bioelectronics, 2014, 53: 167-17.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Tu San Park, Wenyue Li, Katherine E. McCracken and Jeong-Yeol Yoon, "Smartphone Quantifies Salmonella from Paper Microfluidics," Lab on a Chip, 2013, 13(24): 4832-4840.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Tu San Park, Dustin K. Harshman, Christopher F. Fronczek and Jeong-Yeol Yoon,  Smartphone Detection of Escherichia coli from Wastewater Utilizing Paper Microfluidics, The 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2013), Freiburg, Germany, 27-31 October 2013, pp. 1347-1349.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Christopher F. Fronczek, Tu San Park and Jeong-Yeol Yoon, Paper Microfluidic Extraction of Bacterial and Viral Nucleic Acid from Field and Clinical Samples towards a Direct MicroTAS Apparatus, The 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2013), Freiburg, Germany, 27-31 October 2013, pp. 1114-1116.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Dustin K. Harshman, Roberto Reyes, Jeong-Yeol Yoon, Direct Detection of Plasmid-Mediated Antibiotic Resistance in Bloodstream Infection by PCR Using Wire-Guided Droplet Manipulation (WDM), The 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2013), Freiburg, Germany, 27-31 October 2013, pp. 470-472
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Cayla Baynes, Tu San Park, Jeong-Yeol Yoon, Enhanced fluorescent light scatter detection of cancer biomarkers using paper microfluidics, Biosensors 2014: World Congress on Biosensors, Melbourne, Australia, 27-30 May 2014. Paper number O76.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini, Christopher F. Fronczek, Jeong-Yeol Yoon, Rapid, single-step, droplet-based bacterial assay playform on a nanofibrous substrate, Biosensors 2014: World Congress on Biosensors, Melbourne, Australia, 27-30 May 2014. Paper number O112.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Jeong-Yeol Yoon, Paper microfluidics for red wine tasting, Biosensors 2014: World Congress on Biosensors, Melbourne, Australia, 27-30 May 2014. Paper number P3.198.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Phat L. Tran, Daniel A. Martin, Jessica R. Gamboa, Jeong-Yeol Yoon, Marvin J. Slepian, Nanopost fence: a novel strategy of preventing smooth muscle cells topographic migration, Society For Biomaterials (SFB) 2014 Annual Meeting and Exposition, Denver, CO, 16-19 April 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tu San Park, Cayla Baynes, Seong-In Cho, Jeong-Yeol Yoon, Paper microfluidics for red wine tasting, The 9th IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Waikiki Beach, HI, 13-16 April 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini, Celine M. Cohn, Jessica R. Gamboa, Marvin J. Slepian, Xiaoyi Wu, Jeong-Yeol Yoon, Fabrication of a pro-adhesive surface using electrospun PCL nanofibers interspersed with peptide conjugated polystyrene particles, The 9th IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Waikiki Beach, HI, 13-16 April 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Ariana M. Nicolini, Celine M. Cohn, Marvin J. Slepian, Xiaoyi Wu, Jeong-Yeol Yoon, Fabrication of a pro-adhesive surface using electrospun PCL nanofibers interspersed with peptide conjugated polystyrene particles, Annual Meeting of IBE, Lexington, KY, 6-8 March 2014.

Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? 1. With the new collaborative research contract from QIA (Animal, Plant & Fisheries Quarantine & Inspection Agency, South Korea), a handheld polymerase chain reaction (PCR) device has been developed for rapid veterinary diagnostics. My research team (me and two of my graduate students) have visited QIA in June-July 2013 to deliver the alpha prototype and conducted experimental demonstrations. Discussion is currently being made to commercialize this technology. 2. The same technology has been applied to detect blood infections. Two awards have been made, which have started in January 2013, one from AZ Furnace program and the other from Tech Launch Arizona, with the aim of commercializing this technology. A spin-off company, Fast PCR Diagnostics, LLC, has been created, Negotiations are currently being pursued, including option agreement, exclusive practice right, conflict of interest, etc. How have the results been disseminated to communities of interest? Journal paper publications, book publication, conference presentations, and seminar presentations. What do you plan to do during the next reporting period to accomplish the goals? 1. Through the sabbatical leave approved for fall 2013 semester, establish solid collaborations with the top-notch researchers outside the University of Arizona. 2. Participate inmulti-institutional and multi-disciplinary collaborative proposals (including research centerand training grants). 3. Continue to produce top-quality, high-impact peer-reviewed publications.

Impacts
What was accomplished under these goals? 1. Wire-guided droplet PCR technology has been applied for veterinary diagnostics (influenza A) and blood infection, which is very fast and works with blood and tissue samples.A handheld prototype has been developed. 2. A novel method of creating ensemble nanotextured surfaces have been applied to not only endothelial cells, but also fibroblasts and smooth muscle cells. Controllable surfaces, using different types of polymer/nanofiber coatings and electrowetting technology, have also been developed towards creating better cardiovascular devices. 3. A simple paper microfluidics and smartphone-based optical detection is being considered for water quality monitoring (Cryptosporidium and endocrine-disrupting chemicals) and food safety application (Salmonella),

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: David J. You, Tu San Park, Jeong-Yeol Yoon, 2013. Cell-phone-based measurement of TSH using Mie scatter optimized lateral flow assays, Biosensors and Bioelectronics, 40(1): 180-185.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Christopher F. Fronczek, David J. You, Jeong-Yeol Yoon, 2013. Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package, Biosensors and Bioelectronics, 40(1): 342-349.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Jessica R. Gamboa, Samir Mohandes, Phat L. Tran, Marvin J. Slepian, Jeong-Yeol Yoon, 2013. Linear fibroblast alignment on sinusoidal wave micropatterns, Colloids and Surfaces B: Biointerfaces, 104: 318-325.
• Type: Books Status: Published Year Published: 2013 Citation: Jeong-Yeol Yoon, 2013. Introduction to biosensors: from electric circuits to immunosensors, Springer: New York, ISBN: 978-1-4419-6021-4.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Tu San Park, Wenyue Li, Jeong-Yeol Yoon, Paper microfluidics detection of Salmonella using a smart phone, Annual Meeting of IBE, Raleigh, NC, 7-9 March 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Dustin K. Harshman, Roberto Reyes, Tu San Park, Jeong-Yeol Yoon, Extremely fast nucleic acid amplification by droplet manipulation for point-of-care diagnosis of blood infection, Annual Meeting of IBE, Raleigh, NC, 7-9 March 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Jeong-Yeol Yoon, Tu San Park, Wenyue Li, Pei-Shih Liang, Paper microfluidics detection of Salmonella using a smartphone, ASABE Annual International Meeting, Kansas City, MO, 21-24 July, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Pei-Shih Liang, Jeong-Yeol Yoon, Rapid detection of foodborne pathogens within meat utilizing a smartphone biosensor, ASABE Annual International Meeting, Kansas City, MO, 21-24 July, 2013.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Pei-Shih Liang, Jeong-Yeol Yoon, 2013. Optofluidic lab-on-a-chip monitoring of subsurface bacterial transport, Biological Engineering Transactions, 6(1): 17-28.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Katherine E. McCracken, Phat L. Tran, David J. You, Marvin J. Slepian, Jeong-Yeol Yoon, 2013. Shear- vs. nanotopography-guided control of growth of endothelial cells on RGD-nanoparticle-nanowell arrays, Journal of Biological Engineering, 7: 11.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Phat L. Tran, Jessica R. Gamboa, Katherine E. McCracken, Mark R. Riley, Marvin J. Slepian, Jeong-Yeol Yoon, 2013. Nanowell-trapped charged ligand-bearing nanoparticle surfaces - a novel method of enhancing flow-resistant cell adhesion, Advanced Healthcare Materials, 2(7): 1019-1027.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Pei-Shih Liang, Marisa Kleiman, Jeong-Yeol Yoon, Use of biosensor in secondary education curriculum to improve students interest and awareness of science, engineering, and current worldwide issues, ASABE Annual International Meeting, Kansas City, MO, 21-24 July, 2013.

Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: 1. A new collaborative research contract was made to me, from QIA (Animal, Plant & Fisheries Quarantine & Inspection Agency, South Korea), about developing a handheld polymerase chain reaction (PCR) device for rapid veterinary diagnostics. I have visited QIA in May-June 2012 to formulate the collaborative research, and three researchers from QIA have also visit my lab in December 2012 to check our progress. Discussion is currently made to commercialize this technology. 2. The same technology is being applied to detect blood infections. Two awards have been made in December 2012 (both projects start in January 2013), one from AZ Furnace program and the other from Tech Launch Arizona, with the aim of commercializing this technology. A spin-off company, Fast PCR Diagnostics, LLC, has been created in December 2012. Negotiations are currently being made, including option agreement, exclusive practice right, conflict of interest, etc. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Bumsang Kim, Tae Hoon Oh (visiting scholars); David J. You, Tu San Park (post-docs); Jessica R. Gamboa, Pei-Shih Liang, Christopher F. Fronczek, Dustin K. Harshman, Scott V. Angus (graduate students); Katherine E. McCracken, Wenyue Li, Samir Mohandes, Roberto Reyes, Benjamin Erlick, Mariana Yanes, Lily Walsh, Katrina DeCook, Dalton Sycks (undergraduate students). (2) Partner organization: Animal, Plant & Fisheries Quarantine & Inspection Agency (QIA), South Korea. (3) Collaborators: Jae-Young Song (QIA), Marvin J. Slepian (University of Arizona College of Medicine). TARGET AUDIENCES: academic research, food safety, veterinary diagnostics, medical diagnostics, tissue engineering, emerging biotech industry. PROJECT MODIFICATIONS: Nothing significant to report during this repoting period.

Impacts
1. Wire-guided droplet PCR technology have been applied for veterinary diagnostics (influenza A) and blood infection, which is very fast and works with blood and tissue samples. A handheld prototype is currently being developed. A PCT application have been filed. 2. A novel method of creating ensemble nanotextured surfaces have been applied to not only endothelial cells, but also fibroblasts and smooth muscle cells. Controllable surfaces, using different types of polymer/nanofiber coatings and electrowetting technology, are also being developed towards creating better cardiovascular devices. 3. A simple paper microfluidics and smartphone-based optical detection is being considered for water quality monitoring (Cryptosporidium and endocrine-disrupting chemicals) and food safety application (Salmonella).

Publications

• Stemple, C.C., Kwon, H.-J., Yoon, J.-Y. 2012. Rapid and Sensitive Detection of Malaria Antigen in Human Blood with Lab-on-a-Chip. IEEE Sensors Journal, 12(9): 2735-2736.
• Yoon, J.-Y., Kim, B. 2012. Lab-on-a-Chip Pathogen Sensor for Food Safety, Sensors, 12(8): 10713-10741.
• You, D.J., Yoon, J.-Y. 2012. Droplet Centrifugation, Droplet DNA Extraction, and Rapid Droplet Thermocycling for Simpler and Faster PCR Assay Using Wire-Guided Manipulations, Journal of Biological Engineering, 6: 15.
• Angus, S.V., Kwon, H.-J., Yoon, J.-Y. 2012. Field-Deployable and Near-Real-Time Optical Microfluidic Biosensors for Single-Oocyst-Level Detection of Cryptosporidium parvum from Field Water Samples, Journal of Environmental Monitoring, 14(12): 3295-3304.
• Angus, S.V., Kwon, H.-J., Yoon, J.-Y. 2012. Low-Level Detection of Cryptosporidium parvum in Field Water Using Optical Microfluidic Biosensors. Proceedings of SPIE, 2012, 8229: 82290F.
• McCracken, K.E., Tran, P.L., Slepian, M.J., Yoon, J.-Y. 2012. Nanoscale Patterning Under Shear Stress for Guided Endothelial Cell Growth. 2012 Annual Meeting of IBE, Indianapolis, IN, 1-3 March 2012.
• Gamboa, J.R., Tran, P.L., Yoon, J.-Y. 2012. Linear Fibroblast Alignment on Sinusoidal Wave Micropatterns. 2012 Annual Meeting of IBE, Indianapolis, IN, 1-3 March 2012.
• Gamboa, J.R., Yoon, J.-Y., Smith, R.G., Slepian, M.J. 2012. Evaluation of the Performance of a Left Ventricular Assist Device in a Novel In Vitro Heart Failure Model. 2012 Annual Meeting of IBE, Indianapolis, IN, 1-3 March 2012.
• Stemple, C.C., Kwon, H.-J., Yoon, J.-Y. 2012. Rapid and Sensitive Detection of Malaria Antigen in Whole Blood Using a Handheld LOC Device. Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• You, D.J., Tran, P.L., Kwon, H.-J., Yoon, J.-Y. 2012. Extremely Rapid and Fully Reprogrammable Total PCR Assays Using Wire-Guided Droplet Microfluidics. Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• You, D.J., Ehrenkranz, J.R., Yoon, J.-Y. 2012. Cell-Phone-Based Measurement of TSH using Mie Scatter Optimized Lateral Flow Assays. Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• Fronczek, C.F., You, D.J., Yoon, J.-Y. 2012. Single-Pipetting Microfluidic Assay Device for Salmonella in Poultry Package Water. Biosensors 2012: World Congress on Biosensors, Cancun, Mexico, 15-18 May 2012.
• You, D.J., Tran, P.L., Kwon, H.-J., Yoon, J.-Y. 2012. Extremely Rapid and Fully Reprogrammable Total PCR Assays Using Wire-Guided Droplet Microfluidics. 2012 ASABE Annual International Meeting, Dallas, TX, 29 July - 1 August, 2012.
• Fronczek, C.F., You, D.J., Yoon, J.-Y. 2012. Single-Pipetting Microfluidic Assay Device for Salmonella in Poultry Package Water. 2012 ASABE Annual International Meeting, Dallas, TX, 29 July - 1 August, 2012.
• Yoon, J.-Y. 2012. Who We Are & What We Can Do. Resource, 19(3): 19-21.
• Harshmann, D.K., Reyes, R., You, D.J., Yoon, J.-Y. 2012. Device for Near-Instant Diagnosis of Clinical Infection by Convective Droplet Thermocycling and 16s rRNA Hybervariable Region Probes. BMES 2012 Annual Meeting, Atlanta, GA, 24-27 October, 2012.

Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: 3. Develop biological interfaces and sensing systems incorporating microfabrication and nanotechnology. 4. Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Bumsang Kim (visiting scholar); David J. You (post-doc); Hyuck-Jin Kwon, Phat L. Tran, Pei-Shih Liang, Jessica R. Gamboa, Christopher F. Fronczek, C. Christopher Stemple (graduate students); Kenneth J. Geshell, Katherine E. McCracken, Wenyue Li, Christopher J. Love (undergraduate students). (2) Partner organization: Animal, Plant & Fisheries Quarantine & Inspection Agency (QIA), Republic of Korea. (3) Collaborators: Jae-Young Song (QIA), Mark R. Riley (University of Arizona), Marvin J. Slepian (University of Arizona College of Medicine). TARGET AUDIENCES: academic research, food safety, veterinary diagnostics, medical diagnostics, tissue engineering, emerging biotech industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
(1) We have invented a new method of microfluidic droplet manipulation, called wire-guided droplet manipulation, and applied it to the PCR process in detecting H1N1/2009 influenza A and E. coli. 30-cycle thermocycling could be finished in less than 10 min, compared to 1-2 hour with conventional PCR assays. (2) We have designed and manufactured a handheld lab-on-a-chip sensor that detects the presence of E. coli and Salmonella from lettuce, poultry and soil samples. With minimal sample pre-treatment (10% dilution and/or KimWipes paper towel filtration), the level of detection (LOD) was 10 viable cells per 1 mL in the presence of sample matrices. (3) The same device was applied to the detection of airborne pathogens (PRRS and H1N1/2009 influenza A), with 10 pg/mL level of detection in the presence of dust particles. (4) We invented a novel method of creating ensemble nanotextured surfaces and demonstrated enhanced endothelial adhesion and retention under flow. We believe it may be of particular utility for enhancing flow-resistant endothelialization of cardiovascular device.

Publications

• Kwon, H.-J., Angus, S.V., You, D.J., Stemple, C.C., Yoon, J.-Y. 2011. Development of a Handheld Optofluidic Immunosensor to Track the Transport and Distribution of H1N1/2009 Virus in a Mock Classroom, The 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences (uTAS 2011), Seattle, WA, 2-6 October 2011, pp.1421-1423.
• Liang, P.-S., Yoon, J.-Y. 2011. Macroscopic and Microscopic Subsurface Bacterial Transport Model for Soil Bioremediation and Aquifer Contamination with On-site, Real-time, Handheld Lab-on-a-chip Device, ASABE Annual International Meeting, Louisville, KY, 7-11 August 2011.
• Yoon, J.-Y., You, D.J., Geshell, K.J. 2011. Single Cell Level Escherichia coli Detection from Iceberg Lettuce Using a Handheld Microfluidic Immunosensor with Mie Scattering Measurement. ASABE Annual International Meeting, Louisville, KY, 7-11 August 2011.
• Song, J.-Y., Lee, C.-H., Choi, E.-J., Kim, K., Yoon, J.-Y. 2011. Sensitive Mie Scattering Immunoagglutination Assay of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) from Lung Tissue Samples in a Microfluidic Chip. Journal of Virological Methods, 178(1-2): 31-38.
• You, D.J., Geshell, K.J., Yoon, J.-Y. 2011. Direct and Sensitive Detection of Foodborne Pathogens within Fresh Produce Samples Using a Field-Deployable Handheld Device. Biosensors and Bioelectronics, 28(1): 399-406.
• You, D.J., Tran, P.L., Kwon, H.-J., Patel, D., Yoon, J.-Y. 2011. Very Quick Reverse Transcription Polymerase Chain Reaction for Detecting 2009 H1N1 Influenza A Using Wire-Guide Droplet Manipulations, Faraday Discussions, 149(1): 159-170.
• Tran, P.L., Gamboa, J.R., McCracken, K.E., Riley, M.R., Slepian, M.J., Yoon, J.-Y. 2011. Nanowell-trapped Charged Ligand-bearing Nanoparticle Surfaces - a Novel Method for Enhancing Flow Resistant Cell Adhesion, BioInterface Workshop & Symposium, Minneapolis, MN, 24-26 October 2011.
• Tran, P.L., Gamboa, J.R., McCracken, K.E., Riley, M.R., Slepian, M.J., Yoon, J.-Y. 2011. Confluent and Aligned Growth of Endothelial Cells on Nanoparticle Arrays through Focal Adhesion and Endocytotoc Mechanisms. Society For Biomaterials (SFB) Annual Meeting & Exposition, Orlando, FL, 13-16 April 2011.
• Kwon, H.-J., You, D.J., Angus, S.V., Yoon, J.-Y. 2011. Development of a Handheld Lab-on-a-chip Immunosensor to Track the Transport and Distribution of H1N1/09 Virus in a Mock Classroom. Annual Meeting of IBE, Atlanta, GA, 3-5 March 2011.
• Tran, P.L., Gamboa, J.R., McCracken, K.E., Riley, M.R., Slepian, M.J., Yoon, J.-Y. 2011. Confluent and Aligned Growth of Endothelial Cells on Nanoparticle Arrays through Focal Adhesion and Endocytotoc Mechanisms. Annual Meeting of IBE, Atlanta, GA, 3-5 March 2011.

Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: (1) Handheld lab-on-a-chip pathogen sensor: Several different prototypes of a handheld lab-on-a-chip pathogen sensor have been fabricated and tested. Both sensitivity and reproducibility were significantly enhanced under a real biological matrix, by using optofluidic channels. Detection limit of avian influenza A (H3N2) antigens in chicken feces was 1 pg/mL. The same handheld device was also tested for detecting porcine reproductive and respiratory syndrome virus (PRRSV) from the air samples in a real livestock environment and the results not only matches with 30-cycle RT-PCR but also shows better sensitivity. (2) 3-D computational fluid dynamics (CFD) simulation: 3-D CFD model was developed to simulate the spread of virus particles in a livestock environment, and experimentally validated using the above handheld lab-on-a-chip pathogen sensor. (3) Wire-guide polymerase chain reaction (PCR): A newly invented method of wire-guide droplet manipulation was applied for a PCR assay, to speed up the assay time. It took only 6 min and 50 s to complete 30-cycle PCR for H1N1/2009 influenza A virus, using the standard volume of 10 uL that allows us to run gel electrophoresis and gene sequencing, compared to 30 min - 2 hr for conventional PCR assays. (4) Protein array construction: A protein nanoarray made by size-dependent self-assembly (SDSA) successfully detected Octamer-4 (a differentiation factor from human embryonic stem cells) through using FRET as sensing modality. The detection limit was 2 molecules per each bead. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Brian C. Heinze, Phat L. Tran, David J. You, Hyuck-Jin Kwon, Pei-Shih Liang, Jessica R. Gamboa (graduate students); Deepa Patel, Zachary S. Dean, Scott V. Angus, Kenneth Geshell, Katherine E. McCracken (undergraduate students). (2) Partner organization: National Veterinary Research and Quarantine Service, Republic of Korea. (3) Collaborators: Jae-Young Song (National Veterinary Research and Quarantine Service, Republic of Korea), Mark R. Riley (University of Arizona), Marvin Slepian (University of Arizona College of Medicine). TARGET AUDIENCES: academic research, food safety, veterinary diagnostics, medical diagnostics, tissue engineering, emerging biotech industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
(1) The handheld lab-on-a-chip pathogen sensor is currently being tested for detecting pathogenic bacteria from plant tissue samples (lettuce and spinach). Due to its rapidness (<10 min), handheld feature, and greater sensitivity (typically <10 CFU/mL), the device has a strong potential to be widely used for many food safety applications. (2) The same system is also being tested to detect H1N1/2009 in a mock human environment and corresponding 3-D CFD simulation is also being conducted. The simulation model and the handheld pathogen sensor will enable us to improve the ventilation system of a given human environment, as well as to real-time monitor the spread of airborne pathogens in it. (3) The newly invented (patent pending), very quick wire-guide droplet-based PCR system has a strong potential to be used in field applications, including food safety, veterinary diagnostics, medical diagnostics, etc., where a greater sensitivity is warranted. (4) The protein nanoarray is currently being implemented in creating a blood vessel mimic, by patterning a certain combination of receptors in a desired nanoarray geometry within a synthetic vascular graft. So far, vascular endothelial cells are able to create a saturated, monolayered and aligned layer on top of this protein nanoarray, utilizing not only focal adhesion but also endocytosis of nanoparticles that are bound to the nanometer patterns.

Publications

• Kwon, H.-J., Dean, Z.S., Angus, S.V., Yoon, J.-Y. 2010. Lab-on-a-chip for Field Escherichia coli Assays: Long-term Stability of Reagents and Automatic Sampling System. Journal of the Association for Laboratory Automation, 15(3): 216-223.
• Tran, P.L., Gamboa, J.R., You, D.J., Yoon, J.-Y. 2010. FRET Detection of Octamer-4 on a Protein Nanoarray Made by Size-Dependent Self-Assembly. Analytical and Bioanalytical Chemistry, 398(2): 759-768.
• Heinze, B.C., Gamboa, J.R., Kim, K., Song, J.-Y., Yoon, J.-Y. 2010. Microfluidic Immunosensor with Integrated Liquid Core Waveguides for Sensitive Mie Scattering Detection of Avian Influenza Antigens in a Real Biological Matrix. Analytical and Bioanalytical Chemistry, 398(6): 2693-2700.
• Kwon, H.-J., Lee, C.-H., Choi, E.-J., Song, J.-Y., Heinze, B.C., Yoon, J.-Y. 2010. Optofluidic Device Monitoring and Fluid Dynamics Simulation for the Spread of Viral Pathogens in a Livestock Environment. Journal of Environmental Monitoring, 12(11): 2138-2144.
• You, D.J., Tran, P.L., Kwon, H.-J., Patel, D., Yoon, J.-Y. 2011. Very Quick Reverse Transcription Polymerase Chain Reaction for Detecting 2009 H1N1 Influenza A Using Wire-Guide Droplet Manipulations. Faraday Discussions, 149(1): 159-170.
• Heinze, B.C., Yoon, J.-Y. 2011. Nanoparticle Immunoagglutination Rayleigh Scatter Assay to Complement Microparticle Immunoagglutination Mie Scatter Assay in a Microfluidic Device. Colloids and Surfaces B: Biointerfaces, doi:10.1016/j.colsurfb.2011.02.024.

Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: (1) Pathogen detection in lab-on-a-chip: In 2009, we have primarily worked on developing a portable lab-on-a-chip system, through replacing micropositioning stages and optical fibers with on-chip optical waveguides and a miniature spectrometer with an Avalanche photodiode (APD) circuit. External computer was also replaced with a microcontroller board (Arduino) and an LCD display, both powered with two 9-V PP3 batteries. Waveguide chips produced much better reproducibility and sensitivity than our old systems, and successfully tested for the viruses in field samples (nasal swabs from pigs, chicken feces, air samples from swine housing, etc.) with the same impressive detection limits (1 pg/mL avian influenza antigens; 10 TCID50/mL PRRSV). We have also tested E. coli in fresh vegetables with the same system, with detection limit of <100 CFU/mL. (2) Wire-guide droplet microfluidics: This newly conceived method of droplet microfluidics was applied to very quick real-time reverse transcription polymerase chain reaction (vqrRTPCR). The system is capable of amplying the genes extracted from E. coli, in less than 30 min (25 cycles, 1500 bp), which can be further shortened to <5 min (20 cycles, 150 bp). (3) Protein nanoarray construction: A protein nanoarray made by size-dependent self-assembly (SDSA) successfully detected Octamer-4 (a differentiation factor from human embryonic stem cells) through utilizing FRET as sensing modality. This protein nanoarray is currently being implemented in creating a blood vessel mimic (BVM), by patterning a certain combination of receptors/cytokines/growth factors in a desired nanoarray geometry within a synthetic vascular graft. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Brian C. Heinze, Phat L. Tran, David J. You, Hyuck-Jin Kwon, Deepa Patel, and Zachary S. Dean (graduate students); Jessica R. Gamboa, Scott V. Angus, Kenneth Geshell, Teresa Lam (undergraduate students). (2) Partner organization: National Veterinary Research and Quarantine Service, Republic of Korea. (3) Collaborators: Jae-Young Song (National Veterinary Research and Quarantine Service, Republic of Korea), Christopher Y. Choi and Mark R. Riley (University of Arizona). TARGET AUDIENCES: academic research, food safety, veterinary diagnostics, medical diagnostics, tissue engineering, emerging biotech industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
(1) The portable lab-on-a-chip system can be installed in livestock environments to continuously monitor the spread of viral pathogens. This system has a potential to be adapted to human environments, such as a theater, an aircraft cabin or a classroom, to monitor, for example, H1N1 flu. (2) The portable lab-on-a-chip system can also be used routinely in field to monitor contaminations (E. coli or Salmonella) found in leafy vegetables, and potentially other produce as well (ground beef, for example). The wire-guide droplet PCR system can also be made portable to be used for monitoring food safety, which will function as a high-end device (while the waveguide lab-on-a-chip would serve as a low-end device). (3) The protein nanoarray system can become an important tool in designing a better blood vessel mimic, so that tissue-engineered vascular grafts can be used also for small-diameter vessels, hopefully resolving many complications in cardiovascular diseases.

Publications

• Powell, T. B., Tran, P. L., Kim, K., Yoon, J.-Y. 2009. Size-Dependent Self-Assembly of Submicron/Nano Beads-Protein Conjugates for Construction of a Protein Nanoarray. Materials Science and Engineering C, 29(8): 2459-2463.
• Han, J.-H., Kwon, H.-J., Yoon, J.-Y., Kim, K., Nam, S.-W., Son, J.E. 2009. Analysis of the Thermal Environment in a Mushroom House Using Sensible Heat Balance and 3-D Computational Fluid Dynamics. Biosystems Engineering, 104(3): 417-424.
• Yoon, J.-Y., Riley, M.R. 2009. Grand Challenges for Biological Engineering. Journal of Biological Engineering, 3: 16.
• Yoon, J.-Y., Han, J.-H., Choi, C.Y., Bui, M., Sinclair, R.G. 2009. Real-Time Detection of Escherichia coli in Water Pipe Using a Microfluidic Device with One-Step Latex Immunoagglutination Assay. Transactions of the ASABE, 52(3): 1031-1039.
• Han, J.-H., Yoon, J.-Y. 2009. Reusable, Polyethylene Glycol-Structured Microfluidic Channel for Particle Immunoassays. Journal of Biological Engineering, 3: 6.
• Heinze, B.C., Song, J.-Y., Lee, C.-H., Najam, A., Yoon, J.-Y. 2009. Microfluidic Immunosensor for Rapid and Sensitive Detection of Bovine Viral Diarrhea Virus. Sensors and Actuators B: Chemical, 138(2): 491-496.

Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: (1) Protein nanoarray construction: A new method for constructing a protein nanoarray, called size-dependent self-assembly (SDSA) has been further perfected by testing various rinsing protocols and the choice of buffers. Sub-100-nm particles were able to self-assemble on the sub-100-nm patterns created by electron beam lithography, based on the size match. Antibody-antigen binding at the surface of these fluorescent nanoparticles made the particles' fluorescence to be attenuated, a new way of detecting protein binding on a nanoarray format without the need for rinsing. Alternatively, fluorescence resonance energy transfer (FRET) was also demonstrated on the above SDSA-based protein nanoarray using a confocal microscope. This new type of detection provided a better sensitivity and image resolution. Detection was made for Octamer-4 (Oct4), one of the most popular differentiation factors from human embryonic stem cells (hESCs). (2) Pathogen detection with lab-on-a-chip: Extremely low detection limits were demonstrated for Escherichia coli (<10 CFU/mL), bovine viral diarrhea virus (BVDV; <10 TCID50/mL), influenza A H3N2 antigens (<0.1 pg/mL), using forward light scattering detection of latex immunoagglutination assay in lab-on-a-chip. Each assay took <5 min. (3) Wire-guide droplet micro-fluidics: A new way of manipulating droplets on an open surface was demonstrated, called wire-guide droplet microfluidics. Linear and rotational movements, merging, rapid mixing, and optical detection from a droplet were demonstrated. All movements were automated and computerized. Detection limits of latex immunoagglutination assays were largely identical to those with conventional lab-on-a-chip. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Tremaine B. Powell, Jin-Hee Han, Brian C. Heinze, Phat L. Tran, and David J. You (graduate students); Jennine N. Chesler, Anbar Najam, Upasna Chandarana, and Yee Tchao (undergraduate students). (2) Partner organization: National Veterinary Research and Quarantine Service, Republic of Korea. (3) Collaborators: Jae-Young Song (National Veterinary Research and Quarantine Service), Christopher Y. Choi and Gene A. Giacomelli (University of Arizona). TARGET AUDIENCES: academic research, food safety, veterinary diagnostics, medical diagnostics, emerging biotech industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
(1) The SDSA-based protein nanoarray will be used to study endothelialization of a vascular graft, which is already under investigation in our laboratory. (2) Rapid and extremely sensitive lab-on-a-chip utilizing latex immunoagglutinations will be developed as an all-in-one, portable system that can be used to detect pathogens in water distribution systems and in produce (lettuce, spinach, ground beef, etc.) (3) Wire-guide droplet microfluidic system will be applied to further speed up polymerase chain reaction (PCR) to make it a real "real-time" assay (<10 min for 20 cycles).

Publications

• Yoon, J.-Y. (2008). Detection of Avian Influenza Type A H3N2 Virus Antigens in Microchannel and Droplet Microfluidics. Biological Engineering, 1(4): 323-333.
• Kim, K., Yoon, J.-Y., Kwon, H.-J., Han, J.-H., Son, J.E., Nam, S.-W., Giacomelli, G.A., and Lee, I.-B. (2008). 3-D CFD Analysis of Relative Humidity Distribution in Greenhouse with Fog Cooling System and Refrigerative Dehumidifiers. Biosystems Engineering, 100(2): 245-255.
• Yoon, J.-Y. (2008). Open-Surface Digital Microfluidics. The Open Biotechnology Journal, 2: 94-100.
• Han, J.-H., Heinze, B.C., and Yoon, J.-Y. (2008). Single Cell Level Detection of Escherichia coli in Microfluidic Device. Biosensors and Bioelectronics, 23(8): 1303-1306.
• Yoon, J.-Y. (2008). Latex Immunoagglutination Assay in Lab-on-a-Chip. Biological Engineering, 1(1): 79-94.
• Yoon, J.-Y., and You, D.J. (2008). Backscattering Particle Immunoassays in Wire-Guide Droplet Manipulations. Journal of Biological Engineering, 2: 15.
• Heinze, B.C., and Yoon, J.-Y. (2008). Real-Time Monitoring of Fibrinogen Cross-Linking on Model Biomaterial Surfaces with Quartz Crystal Microbalance. The Open Biotechnology Journal, 2: 247-251.

Progress 01/01/07 to 12/31/07

Outputs
OUTPUTS: (1) Through self-assembling antibody-conjugated submicro- or nanoparticles on the desired nanometer wells (through size-matching), we can construct a protein nanoarray capable of single molecule detection. This system will be applied for detecting differentiation factors (e.g. Oct4) in human embryonic stem cell (hESC) cultures, which requires single molecule detection. Antigen-antibody binding will be monitored by using fluorescent particles or NOM (nano-on-micro) configuration. Fluorescent or quantum dot emission would be attenuated if antigen-antibody binding occurs. (2) We will also investigate the use of latex immunoagglutination assays in both microchannel and microdrop lab-on-a-chip platforms, towards automated, reagent-saving and high-throughput pathogen detections. The use of optical light scattering detection should make the detection limit extremely low, close to the single molecule/cell level. Our goal is to make the lab-on-a-chip platform portable, affordable, yet adaptable, through eliminating expensive components and make actuation as simple as possible. PARTICIPANTS: (1) Individuals: Jeong-Yeol Yoon (PI); Keesung Kim (post-doctoral fellow); Lonnie J. Lucas, Tremaine B. Powell, Jin-Hee Han, Brian C. Heinze, Phat L. Tran, and Sarah J. Leung (graduate students); Jennine N. Chesler, and Anbar Najam (undergraduate students). (2) Partner organization: National Veterinary Research and Quarantine Service, Republic of Korea. (3) Collaborators: Christopher Y. Choi (University of Arizona), Jae-Young Song (National Veterinary Research and Quarantine Service), Kye-Seong Kim (Hanyang University), and Gene A. Giacomelli (University of Arizona). TARGET AUDIENCES: Target audiences: academic research, veterinary diagnostics, medical diagnostics, emerging biotech industry.

Impacts
(1) Nanoarray: Fluorescent submicro- and nanoparticles were self-assembled on circular nanometer patterns. Only a single particle could be found in a single circular pattern in most cases. Wells were also patterned as alternating, one at 90-nm and the other at 180-nm. 160-nm blue and 70-nm green fluorescent particles were sequentially added to this pattern. We obtained a fluorescent microscope image (with a 100x oil immersion objective) showing alternating dots of blue and green, successfully demonstrating size-dependent self-assembly. We have conjugated protein molecules to fluorescent, submicro- and nanoparticles and self-assembled them onto nanometer wells. Any biomolecular binding will cause emission attenuation from a fluorescent particle, similar to the NOM concept mentioned below. (2) NOM: We conjugated quantum dots (QDs) onto microspheres to enable multiplex assays as well as to enhance the limit of detection (LOD). We named this configuration nano-on-micro or NOM. Additionally, NOMs are easier to handle than QDs. Since QDs also provide fluorescent emission, we are able to utilize an increase in light scattering for detecting antigen-antibody reaction and a decrease in QD emission to identify which antibody (or antigen) is present. (3) Easy microdrop manipulations: We were able to transport and merge microdrops on a superhydrophobic surface, through utilizing only the difference in the work of adhesion. Neither electrical nor magnetic force was required, unlike the other actuation methods such as electrowetting or magnetofluidics. (4) Single cell detection in lab-on-a-chip: We have built a setup of optical positioning stages for both microchannel and microdrop lab-on-a-chip platforms, and quantified the concentrations of various bacteria and viruses. Detection limits were extremely low; Escherichia coli <10 CFU/mL and Salmonella <5 CFU/mL (single cell level); Bovine viral diarrhea virus (BVDV) <0.5 TCID50/mL and porcine reproductive and respiratory syndrome virus (PRRSV) at 10-4 to 1 TCID50/mL, while that of RT-PCR was only 102 TCID50/mL.

Publications

• Han, J.-H., Heinze, B.C., and Yoon, J.-Y. (2008). Single Cell Level Detection of Escherichia coli in Microfluidic Device, Biosensors and Bioelectronics, doi:10.1016/j.bios.2007.11.013. (in press)
• Lucas, L.J., and Yoon, J.-Y. 2008. On-Chip Detection Using Optical Fibers, In D. Li (ed.), Encyclopedia of Micro- and Nanofluidics, Springer, Berlin Heidelberg. (in press)
• Yoon, J.-Y., and Garrell, R.L. 2008. Biomolecular Adsorption in Microfluidics, In D. Li (ed.), Encyclopedia of Micro- and Nanofluidics, Springer, Berlin Heidelberg. (in press)
• Yoon, J.-Y. (2008). Latex Immunoagglutination Assay in Lab-on-a-Chip, Biological Engineering, 1: 1-16. (in press)
• Heinze, B.C., Song, J.-Y., Han, J.-H., and Yoon, J.-Y. (2008). Latex Immunoagglutination Assay for Bovine Viral Diarrhea Utilizing Forward Light Scattering in Microfluidic Device, Proceedings of SPIE, 6886: 6886-5. (in press)
• Lucas, L.J., Chesler, J.N., and Yoon, J.-Y. (2007). Lab-on-a-Chip Immunoassay for Multiple Antibodies Using Microsphere Light Scattering and Quantum Dot Emission, Biosensors and Bioelectronics, 23: 675-681.
• Kim, K., Giacomelli, G.A., Yoon, J.-Y., Sase, S., Son, J.E., Nam, S.-W., and Lee, I.-B. (2007). CFD Modeling to Improve the Design of a Fog System for Cooling Greenhouses, JARQ, 41: 283-290.
• Yoon, J.-Y., Han, J.-H., Heinze, B., and Lucas, L.J. (2007). Microfluidic Device Detection of Waterborne Pathogens through Static Light Scattering of Latex Immunoagglutination Using Proximity Optical Fibers, Proceedings of SPIE, 6556: 65560M.
• Lucas, L.J., Han, J.-H., Chesler, J., and Yoon, J.-Y. (2007). Latex Immunoagglutination for a Vasculitis Marker in a Microfluidic Device Using Static Light Scattering Detection, Biosensors and Bioelectronics, 22: 2216-2222.
• Han, J.-H., Kim, K.-S., and Yoon, J.-Y. (2007). The Enhanced Diffusional Mixing for Latex Immunoagglutination Assay in a Microfluidic Device, Analytica Chimica Acta, 584: 252-259.

Progress 10/01/06 to 12/31/06

Outputs
Objective 3. To develop nanoscale device and systems Task 1. Development of nano-transducers and bio-recognition system. 50-80 nm AuNPs were synthesized by reducing KAuCl4 with NaBH4. Remaining Cl- ions on the particle surface made these AuNPs negatively charged. Nanometer channels were fabricated by e-beam lithography, on p-doped Si wafer spin-coated with PMMA. E-beam etched the PMMA layer all the way down to the p-doped Si surface, attracting negatively-charged AuNP in spite of the high surface tension inherent in nanostructure. Only 50-nm AuNPs were able to fit the 50-nm nanochannel, whereas 80-nm AuNPs were excluded and eventually rinsed away. AuNPs were conjugated with mouse IgG and self-assembled into this nanochannel. Anti-mouse IgG-FITC was then added to the substrate and the fluorescent microscope identified the binding along with the nanochannel (Figure 2). Thus the proof-of-concept for the proposed protein nanoarray has been demonstrated and these results were published in Biotechnology Progress, 2006. Task 2. Device design. To facilitate the use of QDs for biorecognition, we have conjugated QDs to submicron, core-shell structured latex beads and demonstrated particle immunoagglutination assay in a microfluidic device. We referred this concept as nano-on-micro (NOM), that potentially has the following advantages: easy liquid handling, better particle stability, and the ability to obtain light scattering (from latex beads) and fluorescent emission (from QDs) simultaneously. Upon antigen-antibody binding, the latex beads generated an increase in light scattering (quantitative assay), whereas QDs a decrease in its fluorescent emission (qualitative assay). Since these QDs can be excited at the same wavelength, multiplex assay can be performed in a single chip layout. Task 3. System integration and applications. The proposed protein nanoarray system was planned to be applied for stem cell research, and a collaborative partner was arranged (Kye-Seong Kim's group at Hanyang University, South Korea). Dr. Kim is a world-expert in stem cell research, with state-of-the-art lab facilities supported by the minimal governmental restriction on the use of human embryonic stem cell (hESCs).

Impacts
Objective 3. To develop nanoscale device and systems Task 1. Development of nano-transducers and bio-recognition system. - Nanoarray: IgGs will be patterned onto not nanochannels but nanowells. Each array grid will hold a single AuNP or QD and subsequently single IgG. QDs will also be tested for size-dependent assembly and subsequently fluorescent biorecognition. NOM concept will also be applied to nanoarray system. - Lab-on-a-chip: NOM concept will also be applied to microfluidic device. Task 2. Device design. - Nanoarray: 10-30 array grids will be constructed. - Lab-on-a-chip: Multiplex assay will be performed within a single microchannel, using NOM concept in microfluidic device. Task 3. System integration and applications. - Nanoarray: A key differentiating factor, Oct4 and Cdx2, will be detected from a hESC culture, using the nanoarray system developed above. - Lab-on-a-chip: A disease marker from human or animal blood serum will be detected using the above NOM concept in microfluidic device.

Publications

• Lonnie J. Lucas, Jin-Hee Han, Jenine Chesler, Jeong-Yeol Yoon. 2007. Latex immunoagglutination for a vasculitis marker in a microfluidic device using static light scattering detection. Biosensors and Bioelectronics, in press. doi:10.1016/j.bios.2006.10.029
• Jeong-Yeol Yoon, Jin-Hee Han, Brian Heinze, Lonnie J. Lucas. 2007. Microfluidic device detection of waterborne pathogens through static light scattering of latex immunoagglutination using proximity optical fibers. Proceedings of SPIE, 6556: 655619, in press.
• Jin-Hee Han, Kye-Seong Kim, Jeong-Yeol Yoon. 2007. The enhanced diffusional mixing for latex immunoagglutination assay in a microfluidic device. Analytica Chimica Acta, 584: 252-259.
• Lonnie J. Lucas, Jin-Hee Han, Jeong-Yeol Yoon. 2006. Using highly carboxylated microspheres to simplify immunoassays and enhance diffusional mixing in microfluidic devices. Colloids and Surfaces B: Biointerfaces, 49: 106-111.
• Tremaine Powell, Jeong-Yeol Yoon, 2006. Fluorescent biorecognition of gold nanoparticle - IgG conjugates self-assembled on e-beam patterns. Biotechnology Progress, 22: 106-110.