Conference Registration, Materials Pick-Up, Morning Coffee and Breakfast Pastries

Welcome by Dr. Johnsee Lee, Chairman, Taiwan Bio Industry Organization

Accelerating the Detection Speed of Circulating DNA/RNA by Electrokinetic Concentration in a Microfluidic Chip
Yong-Ak (Rafael) Song, Assistant Professor of Mechanical and Biomedical Engineering, New York University Abu Dhabi, United Arab Emirates

The cell-free DNA (cfDNA), which consists of fragments of DNA from dying cancer cells, and exosomes released during fusion of the multivesicular endosomes have demonstrated great promise as an alternative biomarker for cancer detection and assessment of treatment. To enhance the detection sensitivity and speed of “Liquid Biopsy” tools for cfDNA and exosomal RNA, we have developed a microfluidic concentrator chip based on ion concentration polarization (ICP) that can preconcentrate target nucleic acids electrokinetically from a ~µL fluidic sample into a ~pL plug in the vicinity of the molecular capture probes. ICP preconcentration not only reduces the analyte diffusion length but also increases the binding reaction rate, and as a result, ICP-enhanced biosensors allow much faster hybridization than standard diffusion-limited biosensors such as microarrays. Utilizing an ion-selective conductive polymer membrane, poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS), in a microfluidic channel, we could achieve a concentration increase of DNA by 6 orders of magnitude from an initial concentration of 100 fM at 50V within 10 min. As for the detection via surface hybridization on morpholino (MO) probes, a class of uncharged DNA mimics, DNA target concentration as low as 10 pM was detected within 15 min. In the case of an exosomal RNA, miR21, we could accelerate its surface hybridization on MO probes by 7-fold using the electrokinetic concentrator. It even allowed an increase of the concentration of exosomes (from human breast cancer cell lines, ~105/mL) locally for a sensitive fluorescence detection. These results demonstrate a potential application of the microfluidic electrokinetic concentrator for a rapid detection of cfDNA and exosomal RNA molecules down to few pico- and femtomolar regimes.

Coffee and Tea Break and Networking in the Exhibit Hall

Microfluidic Platform For Personalized Medicine and Healthcare
Nien-Tsu Huang, Assistant Professor, Department of Electrical Engineering, National Taiwan University, Taiwan

A discussion regarding how to develop a fully integrated microfluidic platform enabling on-chip sample preparation and in-situ biomarkers detection. The platform can potentially be applied for personalized healthcare.

Technology Spotlight:
All in One – Advanced Technologies for Complex Low Cost Microfluidic Devices in Glass, Silicon and Quartz
Klaus Kadel, Business Development, Little Things Factory GmbH

The Little Things Factory covers the whole portfolio to set up new functionalities for microfluidic systems in glass and we describe recent innovations in this field. His talk will introduce new possibilities to combine microfluidics with hermetically sealed electrical connectors for various applications that require electrodes in contact with fluids or electrodes isolated from fluids.

Networking Lunch in the Exhibit Hall -- Meet the Exhibitors and View Posters

Liquid Crystals in Microfluidic Devices for Biosensing Applications
Kun-Lin Yang, Associate Professor, Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore

Microfluidic sensors possess several advantages such as smaller sample volume, shorter assay time and lower cost compared to conventional assays performed in 96-well plates. However, because of its miniaturized format, microfluidic sensors also faced serious challenges in signal detection because of inherent short light path length, small sample volume, and short residence time. To overcome these limitations in microfluidic sensors, ultrasensitive signal detection and amplification mechanism are required. In this presentation, integration of liquid crystal (LC) materials with microfluidic devices for various chemical and biological sensing applications will be discussed. Because molecules form a LC phase have polar functional groups which have long-range interactions, they respond collectively to surface molecular binding events and change their orientations accordingly. This working principle has been exploited to develop highly sensitive biosensors in microfluidic devices. Using LC for signal transduction and amplification is advantageous because it is highly sensitive, fast and simple. More importantly, optical outputs of LC can be observed with the naked eye directly without using any complex instrumentation. Interference colors of LC also offer a simple way for qualitative and spatial analysis of protein concentration in microfluidic devices. Using LC-based microfluidic sensors, different biomolecules such as antibodies and enzymes can be detected in microfluidic devices.

Miniaturized Devices for Point-of-Care Diagnostic Applications
Chien-Fu Chen, Assistant professor, Institute of Applied Mechanics, National Taiwan University, Taiwan

A discussion regarding the miniaturized systems utilizing micro/nanomaterials for biomedical point-of-care sensing applications.

Rapid Detection of Pathogenic Bacteria on a Microfluidic Disc
Izumi Kubo, Professor, Soka University, Japan

We have developed an originally designed microfluidic disc-like device with microchannels and microchambers for single cell isolation. For the detection of genes in isolated cells on the device, hot cell-direct polymerase chain reaction (PCR) method was invented. In this study, food pathogenic bacteria such as Salmonella enterica was isolated on the disc and determined by the amplification of its specific gene based on hot cell-direct PCR using fluorescent probe. Relative fluorescence of each microchamber after/before PCR was determined to detect the bacteria. By the method, rapid and simple detection of bacterial cells was performed.

Afternoon Coffee and Tea Break and Networking in the Exhibit Hall

Ultra Low-Cost Smartphone Optosensors for Mobile Point-of-Care Diagnostics
Lei Li, Assistant professor, School of Mechanical and Materials Engineering, Washington State University, United States of America

We have developed a series of opto-sensing platforms that cover a wide range of applications. Especially, we have developed an ultra low-cost miniaturized multichannel smartphone opto-sensing platform with high accuracy and sensitivity. Our unique opto-mechanical system design and manufacturing process made our platforms appropriate for low-cost in-field applications.

PeptaSensors Open up New Avenues For POC Diagnostics
Manish Biyani, Associate Professor, Japan Advanced Institute of Science and Technology, Japan

This talk will introduce peptide aptamer-based biosensor, so-called peptasensor, for making undetectable sensing possible for routine and fast-tracking molecular diagnostics. Small peptide chains, which can arrange themselves into tridimensional functional structures, exhibit a smaller binding footprint allowing for a more thorough and precise interrogation of the target molecule. We engineered these peptide chains by an iterative process of in vitro selection procedure and applied to recognize certain biomarkers with high affinity and specificity. The applicability of these peptide aptamer to fabricate the new age of integrated electronic biosensors to detect unreachable target molecules will be addressed in this presentation.

Close of Day 1 of the Conference

Morning Coffee, Tea and Networking

Morning Coffee, Tea and Networking in the Exhibit Hall

Flexible Microfluidic Control for Highly Functionalized LOAC Platform
Masahiro Motosuke, Professor, Department of Mechanical Engineering, Tokyo University of Science, Japan

In this talk, I would like to introduce some our recent work in the development of sophisticated microfluidic control by using external-field for on-demand functionality in LOAC platform.

Development of PQQ-Glucose Dehydrogenase(GDH)-based Biosensors
Zhong Guo, Research Officer, Institute for Molecular Bioscience, The University of Queensland, Australia

Recently, biosensors have become a practical alternative to complex and expensive analytical instruments used in healthcare, agriculture and environmental monitoring. Among several currently used detection technologies such as optical, acoustic and piezoelectric, electrochemical sensors feature prominently due to their simplicity, specificity and high performance. Electrochemical blood glucose sensors are the most commercially successful biosensors accounting for 60% of global biosensor market. The sensors are based on amperometric monitoring of glucose oxidation by recombinant glucose dehydrogenase (GDH). We have developed biosensors by engineering of GDH through insertion and splitting strategies. The sensors can be adopted for the detection of Calcium ion, immunosuppressant drugs, salivary a-amylase protein or protease activity of thrombin and Factor Xa. We expect that the sensor architectures could be expanded to the detection of other biochemical activities, posttranslational modifications, nucleic acids and inorganic molecules. And we also expected the sensors can be used for the developing of Point-of Care diagnostics. Particularly, clinic testing of our first Point-of-Care device developed for Calcium detection for human blood, saliva and urine samples have been proceeded recently.

Induction of Stem Cell Self-Assembly by Adhesion Restriction
Kennedy Okeyo, Senior Lecturer, Institute for Frontier Life and Medical Sciences, Kyoto University, Japan

We have recently developed a novel culture technique, namely, the shoji technique, which involves the use of micro-structured large porous meshes as substrates for cell culture and growth in a suspension under restricted adhesion condition. This talk will highlight our on-going work on stem cell culture by the shoji technique, specifically, how this results in induction of self-assembly organization and, ultimately, differentiation. We show that the latter occurs mechanically and results in dynamic morphogenetic transformations that yields unique 3D cysts from originally 2D stem cell sheets. Overall, we show that the shoji technique presents a novel approach to modulate the cell culture microenvironment, resulting in mechanical induction of directed stem cell differentiation.

PERSONAL PHOTOMETER “PHOTOPETTE” WITH “LAB-IN-A-TIP” TECHNOLOGY FOR LIFE-SCIENCE APPLICATIONS
Dieter Trau, Associate Professor, University of Singapore, Singapore

Here, we introduce two novel concepts, a personal photometer “PHOTOPETTE” combined with “Lab-in-a-Tip” technology. The “Lab-in-a-Tip” is based on colorimetric reactions of an analyte with a reagent incorporated into a disposable polymer tip. The “PHOTOPETTE” is used for optical read out. Less sample volume required and on-spot detection are the advantages of this technology. We demonstrated pH and protein measurements in our “Lab-in-a-Tip design”. The work was conducted in collaboration with Tip Biosystems Pte Ltd. (This work is supported by National Research Foundation Singapore, NRF2014NRF-POC002-045).

Networking Lunch in the Exhibit Hall -- Meet the Exhibitors and View Posters

Microfluidic Cell Culture Devices to Control Gaseous Microenvironments in vitro
Yi Chung Tung, Associate Research Fellow,Research Center for Applied Sciences, Academia Sinica, Taiwan

Introduction of microfluidic devices capable of controlling various gaseous concentrations and gradients for in vitro cell culture developed in my laboratory.

Binding Kinetic Constants Measurement by Using Fiber Optic Particle Plasmon Resonance Biosensor
Shau-Chun Wang, Professor and Director, Department of Chemistry and Biochemistry and the Center for Nano BioDetection, National Chung Cheng University, Taiwan

Biosensing methods have been a popular means to measure the binding affinity and rate constants of immuno-logical reactions. In particular, the quantitative investigation of binding kinetics provides important information regarding the interactions between the docking target molecule and the counterpart probe molecule. Fiber optic particle plasmon resonance (FOPPR) biosensor, one simple and label-free sensing platform, has been successfully used to determine binding kinetic constants. The FOPPR biosensor is based on an optical fiber modified on gold nanoparticles, where the surface of gold nanoparticles has been conjugated by a mixed self-assembled monolayer (SAM) and a molecular probe reporter to dock with the corresponding analyte species. The binding process is recorded in static mode when analyte solution is loaded in one static sensing cell. The ability to analyse biomolecules without flow may be especially useful when an experiment only has a small amount of analyte available to characterize a biomolecular interaction. We compare FOPPR biosensor with commercialized Biacore 3000 SPR biosensor to determine the binding kinetic constants, which is limited by analyte injection flow rate and the probe concentration on the chip in Biacore 3000 SPR biosensor whereas FOPPR is not affected by these two effects. In addition, we established the general mathematical model to estimate the binding kinetic rate constants based on the pseudo-first-order model. We will discuss the agreement of obtained association rate constants (ka) and dissociation rate constants (kd) of anti-ovalbumin with ovalbumin and IgG binding with anti-IgG, between the computational results using our mathematical model, and the measured values.

Direct Observation of Folding Stability of Individual Chromosomes Isolated From Single Mammalian Cells in a Microfluidic Channel
Hidehiro Oana, Associate Professor, The University of Tokyo, Japan

Chromosomes were isolated from single mammalian cells in a microfluidic channel. Then, the isolated chromosomes were tethered at microstructures and exposed to high-salt solution in the channel. As a result, the chromosomes were unfolded non-uniformly, which can provide insights into epigenetic mechanisms.

Highly-Sensitive Label-Free Optical Biosensors and Multiplex Immuno-Magnetic Platform for Point-of-Care Diagnostics
Petr Nikitin, Professor and Head of Laboratory, Prokhorov General Physics Institute, Russian Academy of Sciences, Russian Federation

Several types of original biosensors will be presented. Intelligent biosensing systems are developed based on nanoparticles that can employ molecular interactions to perform any Boolean logic function. These systems are efficiently used for logic-gated biosensing with lateral flow test-strips and 3D filters. Besides, multichannel label-free biosensors are designed based on the spectral correlation interferometry (SCI) for detection of various analytes by measuring changes in thickness of a biolayer on functionalized glass slips used as affordable single-use sensor chips. The SCI method is not sensitive to bulk refractive index of a solution under test and provides signals in metrological units (pm or nm). Using real-time monitoring of bioreactions by the SCI, a new multiplex dry-reagent immunomagnetic (DRIM) biosensing platform is developed for rapid high-precision quantitative analyses of complex mediums. It is based on the highly sensitive magnetic particle quantification (MPQ) method that provides detection limit of 60 zeptomoles or 0.4 ng of magnetic nanolabels and extremely wide linear dynamic range of 7 orders. The DRIM platform has permitted the detection in human serum of as low as 25 pg/ml total prostate specific antigen during 30-min assay. The featured 3-fold signal increase per every order of concentration within 3.5 orders of magnitude allows precise analysis of antigen concentrations in a wide range. In addition, based on the MPQ technique, a new cytometry method has been developed and successfully tested for rapid assessment of cancer HER2/neu status of cells, for estimation of expression level of various antigens on cell surfaces and cancer diagnostics.

Single Cell Clinical Enzyme Analysis for Precision Medicine by Using Continuous Flow Microfluidics
Chia Hung Chen, Assistant Professor, National University of Singapore, Singapore

Precision medicine refers to giving the right therapeutics, to the right patient, at the right time. In the context of cancer, successful implementation of precision medicine, requires treatment individualization not only taking into account patient and tumor factors, but also tumor heterogeneity and tumor evolution over time. In this study, a continuous flow microfluidic device was developed as a functional flow cytometer (Droplet FACS) to detect secreted multiplexed protease activities at single cell resolution. The individual cells from patient samples are encapsulated within water-in-oil droplets for single cell multiplexed protease assay. We modified FRET (fluorescence resonance energy transfer)-based substrates to accommodate different fluorescent pairs with distinct excitation and emission wavelengths to obtain multiple signals from droplets containing single cells. Four substrate-protease reactions in a droplet were simultaneously monitored at three distinct pairs of fluorescent excitation (UV: 400nm, B: 470nm, G: 546nm, R: 635nm) and emission (B: 520nm, G: 580nm, R: 670nm) wavelengths. To infer a quantitative profile of multiple proteolytic activities from single cells, we applied the computational method Proteolytic Activity Matrix Analysis (PrAMA). The capability to determine multiple protease activities at single cell resolution has the potential to characterize tumor progress of individual patients.

Saving Lives With Brownian Motion
Han-Sheng Oswald Chuang, Associate Professor, Department of Biomedical Engineering, National Cheng Kung University, Taiwan

Sepsis is a fatal infectious disease claiming thousands of lives every year. Antimicrobial susceptibility testing (AST) plays a pivotal role in the success of treatments. However, the turnaround time for the outcome of conventional AST usually requires over 24 hours, resulting in high patient mortality. Moreover, antibiotics abuse can also incubate the booming of superbugs. A reliable and efficient drug screen becomes increasingly important to date to save lives in a timely fashion. To this end, a technique combining optical diffusometry and bead-based immunoassays is developed herein to achieve a rapid quantification of target microorganisms. The limit of detection of the method could achieve 100 CFU/mL. This study provides valuable information to timely treatments against poly-microbial diseases in the near future.

Microfluidics-assisted Spinning of Single Micrometer-sized Protein Microfibers Using Alginate as Sacrificial Matrix
Hisataka Hiramatsu, Graduate Student, Chiba University, Japan

We propose a facile and versatile approach to fabricate proteins microfibers using a microfluidic spinning system and sacrificial matrices of alginate. We prepared single micrometer-width gelatin and albumin fibers and applied them as solid scaffolds for 3D mammalian cell cultivation.

Close of Day 2 of the Conference