Welcome, Introduction, Topics Addressed and Conference Opening by Conference Chairperson
Leanna Levine, Founder & CEO, ALine, Inc., United States of America

Microfluidics for Interrogating Intact-Tumor Biopsies
Albert Folch, Professor of Bioengineering, University of Washington, United States of America

Cancer remains a major healthcare challenge worldwide. It is now well established that cancer cells constantly interact with fibroblast cells, endothelial cells, immune cells, signaling molecules, and the extracellular matrix in the tumor microenvironment (TME). Present tools to study drug responses and the TME have not kept up with drug testing needs. The number of clinical trials of combination therapies has been climbing at an unsustainable rate, with 3,362 trials launched since 2006 to test PD-1/PD-L1-targeted monoclonal antibodies alone or in combination with other agents. We have developed a microfluidic platform (called Oncoslice) for the delivery of multiple drugs with spatiotemporal control to live tumor biopsies, which retain the TME. We have developed the use of Oncoslice for the delivery of small-molecule cancer drug panels to glioblastoma (GBM) xenograft slices as well as to slices from patient tumors (GBM and colorectal liver metastasis). In addition, we have developed a precision slicing methodology that allows for producing large numbers of cuboidal micro-tissues (“cuboids”) from a single tumor biopsy. We have been able to trap cuboids in arrays of microfluidic traps in a multi-well platform. This work will potentially allow for the high-throughput application of drugs to intact human tumor tissues.

Microfluidic Technologies For Liquid Biopsy
Daniel Chiu, A. Bruce Montgomery Professor of Chemistry, University of Washington, United States of America

This presentation will describe microfluidic technologies we developed for liquid biopsy and precision medicine. Specifically, a rare-cell isolation instrument we call eDAR (ensemble decision aliquot ranking), a nanofluidic technology for the high-sensitivity sorting of exosomes, and a digital-nucleic-acid detection and analysis platform based on our SD (self-digitization) chip. I will outline the workings of these microfluidic platforms, describe their performance, and discuss the clinical questions we are addressing with these technologies.

Precision Biology: Deep Profiling of Single Cells Using Electrophoretic Cytometry
Amy Herr, Professor, University Of California Berkeley, United States of America

Underpinning single-cell measurement tools, microfluidic design offers the throughput, multiplexing, and quantitation needed for rich, multi-dimensional data. Genomics and transcriptomics are leading examples. Yet, while proteins are the dynamic, downstream effectors of function, the immunoassay remains the de facto standard (flow cytometry, mass cytometry, immunofluorescence). We posit that to realize the full potential of high-dimensionality cytometry, new approaches to protein measurement are needed. I will describe our ‘electrophoretic cytometry’ tools that increase target selectivity beyond simple immunoassays. Enhanced selectivity is essential for targets that lack high quality immunoreagents – as is the case for the vast majority of protein forms (proteoforms).  I will share our results on highly multiplexed single-cell western blotting and single-cell isoelectric focusing that resolves single charge-unit proteoform differences. In fundamental engineering and design, I will discuss how the physics and chemistry accessible in microsystems allows both the “scale-down” of electrophoresis to single cells and the “scale-up” to concurrent analyses of large numbers of cells. Precise reagent control allows for integration of cytometry with sophisticated sample preparation – the unsung hero of measurement science. Lastly, I will link our bioengineering research to understanding the role of protein signaling and truncated isoforms in development of breast cancer drug resistance and understanding protein signaling in individual circulating tumor cells. Taken together, we view microfluidic design strategies as key to advancing protein measurement performance needed to address unmet gaps in quantitative biology and precision medicine.

Microengineering a Physiologic Colon Replica
Nancy Allbritton, Frank and Julie Jungers Dean of the College of Engineering and Professor of Bioengineering, University of Washington in Seattle, United States of America

Organ-on-chips are miniaturized devices that arrange living cells to simulate functional subunits of tissues and organs. These microdevices provide exquisite control of tissue microenvironment for the investigation of organ-level physiology and disease. A 3D polarized epithelium using primary human gastrointestinal stem cells was developed to fully recapitulate gastrointestinal epithelial architecture and physiology. A planar monolayer comprised of stem/proliferative and differentiated primary cells is cultured on a shaped hydrogel scaffold with an array of crypt-like structures replicating the intestinal architecture. These planar layers display physiologic drug transport and metabolism and immunologically appropriate responses. Facile co-culture with other cell types such as immune cells or myofibrolasts is readily achieved. A dense mucus layer is formed on the luminal epithelial surface that is impermeable to bacteria and acts a barrier to toxins. The in vitro mucus has remarkably similar in its biophysical properties to that produced in vivo. Imposition of chemical gradients across the crypt long axis yields a polarized epithelium with a stem-cell niche and differentiated cell zone. The stem cells proliferate, migrate and differentiate along the crypt axis as they do in vivo. An oxygen gradient across the tissue mimic permits luminal culture of anaerobic bacteria while maintaining an oxygenated stem cell niche. This in vitro human colon crypt array replicates the architecture, luminal accessibility, tissue polarity, cell migration, and cellular responses of in vivo intestinal crypts. This bioanalytical platform is envisioned as a next-generation system for assay of microbiome-behavior, drug-delivery and toxin-interactions with the intestinal epithelia.

Workflow Automation with Digital and Customizable Microfluidics: Challenges and Opportunities
Anita Rogacs, Head of Life Sciences Strategy and R&D, HP Labs, United States of America

Life Sciences analytical workflows benefit greatly from microfluidic automation, leading to improvement in accuracy, multiplexing, efficiency, and cost. Yet microfluidics has faced significant barriers to adoption in both diagnostics and drug discovery, due to constrained designs and long and costly development pipelines. HP is addressing this challenge by leveraging our digital and customizable mode of microfluidics, eliminating the need to design a “new chip” for every new workflow, target and reagent.

Rapid Sensitive and Simple NAAT Self-testing for SARS-CoV-2 with Paper-based Microfluidics
Paul Yager, Professor, Department of Bioengineering, University of Washington, United States of America

Whether to guide treatment of an individual’s infection, or to manage an ongoing pandemic, or to prevent the next one, there is an urgent need for low-cost rapid diagnostic devices capable of identifying the cause of infectious disease that work wherever the person is, not just in a centralized laboratory.  “Ubiquitous diagnostics” can bring the best diagnostic capabilities to homes, physicians’ office laboratories and pharmacies in the developed world, or to places in the developing world where nothing is available now.  We had been working to develop simple single-use instrument-free devices for nucleic acid amplification tests (NAATs) for the presence of respiratory diseases, Chlamydia and gonorrhea, Mycobacterium tuberculosis and HIV.  To enable these devices, we have created a suite of stand-alone or integrated sample-handling components that can process blood, urine or swabs.  With the outbreak of the COVID-19 pandemic, we have pivoted to detection of the SARS-CoV-2 virus to address the unmet need for self-testing in the home at sensitivity levels that would allow detection of presymptomatic cases.  We will show recent progress in reducing such processes to a few user-friendly steps appropriate for untrained users.  We have established UbiDX to commercialize the SARS-CoV-2 test and multiplexed panels of pathogens for use in self-testing in a variety of scenarios.

Resources to Aid Point of Care Developers to Market
Joany Jackman, Senior Scientist, Technical Lead, Technology Development Core, Johns Hopkins Center for Point-of-Care Technologies Research for Sexually Transmitted Diseases, The Johns Hopkins University Applied Physics Laboratory, United States of America

The Johns Hopkins Center for Point-of-Care Technologies Research for Sexually Transmitted Diseases (the Center) is part of the National Institute of Biomolecular Imaging and Bioengineering’s Point of Care Technologies Research Network (POCTRN).  Established in 2007, this network of centers was created to help developers of promising technologies reach the market faster and for less money. This is done by pairing them with scientists, clinicians and laboratorians who are experts in the disease targets for which they are developing their devices.   As part of this program, our Center provides expert guidance in the biology of STDs, clinicians’ view of diagnostics, clinical samples at low or no cost, clinical evaluations, tactical funding, and now systems engineering. Recently rapid development for COVID diagnostics (RADx) has been added to the mission of the POCTRN Centers. A description of ways to interact with our Center will be highlighted.

Microneedle-Electrochemical Sensors: Towards a Lab Under the Skin
Joseph Wang, Chair of Nanoengineering, SAIC Endowed Professor, Director at Center of Wearable Sensors, University of California-San Diego, United States of America

Wearable sensors have received a major recent attention owing to their considerable promise for monitoring the wearer’s health and wellness. The medical interest for wearable systems arises from the need for monitoring patients over long periods of time. These devices have the potential to continuously collect vital health information from a person’s body and provide this information to them or their healthcare provider in a timely fashion. Such sensing platforms provide new avenues to continuously and non-invasively monitor individuals and can thus tender crucial real-time information regarding a wearer’s health.

This presentation will focus on our recent efforts for using microneedle sensor arrays for simultaneous minimally-invasive real-time monitoring of multiple biomarkers. Owing to the arrayed nature of the microneedle structures, various target analytes can be detected at different individually-addressable microneedles for realizing such multiplexed sensing operation. The preparation and characterization of such wearable electrochemical microneedle sensor arrays will be described, along with their current status, advantages, latest applications, and future prospects and challenges.

Nanobiosensors for Diagnostics Applications
Arben Merkoçi, ICREA Professor and Director of the Nanobioelectronics & Biosensors Group, Institut Català de Nanociencia i Nanotecnologia (ICN2), Barcelona Institute of Science and Technology (BIST), Spain

There is a high demand to develop innovative and cost effective devices with interest for health care beside environment diagnostics, safety and security applications. The development of such devices is strongly related to new materials and technologies being nanomaterials and nanotechnology of special role. We study how new nanomaterials such as nanoparticles, graphene, nano/micromotors can be integrated in simple sensors thanks to their advantageous properties. Beside plastic platforms physical, chemical and mechanical properties of cellulose in both micro and nanofiber-based networks combined with their abundance in nature or easy to prepare and control procedures are making these materials of great interest while looking for cost-efficient and green alternatives for device production technologies. Both paper and nanopaper-based biosensors are emerging as a new class of devices with the objective to fulfil the “World Health Organization” requisites to be ASSURED: affordable, sensitive, specific, user-friendly, rapid and robust, equipment free and deliverable to end-users. How to design simple paper-based biosensor architectures? How to tune their analytical performance upon demand? How one can couple nanomaterials such as metallic nanoparticles, quantum dots and even graphene with paper and what is the benefit?   How we can make these devices more robust, sensitive and with multiplexing capabilities? Can we bring these low cost and efficient devices to places with low resources, extreme conditions or even at our homes? Which are the perspectives to link these simple platforms and detection technologies with mobile communication? I will try to give responses to these questions through various interesting applications related to protein, DNA and even contaminants detection all of extreme importance for diagnostics, nanotheranostics, environment control, safety and security.

Immunodiagnostic Point-of-Care Testing (POCT) for Rapid Serological Tests of COVID-19 Exposure and Immunity
Chong Ahn, Distinguished University Research Professor, Mitchell P. Kartalia Chair Professor of BioMEMS, University of Cincinnati, United States of America

A new compact immunodiagnostic point-of-care testing (POCT) analyzer which can rapidly (within 10-15 minutes) and accurately detect anti-SARS-COV-2 IgG and IgM antibodies has been explored and developed, in combination with a neutralization assay which measures neutralizing antibodies to SARS-CoV-2, with the goal of linking serological diagnostics to immune status of infected individuals.

The compact POCT analyzer platform with a new multiplex microchannel-based capillary flow assay (MCFA) lab chip can perform a rapid, real time measurement of anti-SARS-COV-2 IgM and IgG in whole blood for determining COVID-19 exposure and immunity. The POCT platform consists of (a) a multiplex polymer lab-on-a-chip (LOC) that can successfully integrate multiple features of sandwich ELISA and lateral flow tests to implement MCFA using on-chip dried reagents and (b) a portable compact fluorescent analyzer to simultaneously detect multiple targets from the multiplexing immunoassay LOC.  In addition, the use of a monoclonal antibody specific for the receptor binding domain (RBD) of the S-protein would enable the utilization of the MCFA device for a competitive assay to determine the presence of neutralizing antibody in the patient sample, leading to an ideal POCT that not only detects COVID-19 exposure but can also confer immune status.

This serological diagnostics in a compact POCT analyzer platform will provide a new paradigm for rapid and cost-effective diagnosis of rapidly-transmitting diseases like COVID-19.