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Welcome and Introduction by Conference Chairperson: Emerging Themes in Microfluidics 2023
カンファレンス議長による挨拶：マイクロフルイディクスにおける新たなテーマ2023年
Claudia Gartner, CEO, microfluidic ChipShop GmbH, Germany

Nanobiodevices, Quantum Life Science, and AI for Future Healthcare
ナノバイオデバイス、量子生命科学、および未来のヘルスケアに向けたAI
Yoshinobu Baba, Professor, Nagoya University, Japan

We have investigated nanobiodevices, quantum life science, and AI for biomedical applications and healthcare.  Nanowire devices are extremely useful to isolate extracellular vesicles from body fluids and vesicle-encapsulated microRNA analysis.  The device composed of a microfluidic substrate with anchored nanowires gives us highly efficient collections of extracellular vesicles in body fluids and in situ extraction for huge numbers of miRNAs (2,500 types) more than the conventional ultracentrifugation method. Nanowire devices gave us the miRNA date for several hundred patients and machine learning system based on these miRNA data enabled us to develop the early-stage diagnosis for lung cancer, brain tumor, pancreas cancer, liver cancer, bladder cancer, prostate cancer, diabetes, heart diseases, and Parkinson disease.  Nanowire-nanopore devices combined with AI (machine learning technique) enable us to develop mobile sensors for SARS-CoV-2, PM2.5, bacteria, and virus in the environment. MEXT Quantum Leap Flagship Program (Q-LEAP), which I lead, has been developing biological nano quantum sensors, quantum technology-based MRI/NMR, and quantum biology and biotechnology. Nanodiamonds, with nitrogen-vacancy centers, and quantum dots are applied to develop quantum sensors for quantum switching intra vital imaging for iPS cell (induced pluripotent stem cells) based regenerative medicine, and quantum photo immuno-therapeutic devices for cancer. Multifunctional quantum nanoparticles were developed for fluorescence/MR bimodal in vivo imaging-guided potothermal-intensified chemodynamic synergetic therapy of targeted tumors.

Microfluidic Wearable Sensors for Healthcare and Health Metaverse Applications
ヘルスケアとヘルスメタバースアプリケーションのためのマイクロ流体ウェアラブルセンサー
Chwee Teck Lim, NUS Society Chair Professor, Department of Biomedical Engineering, Institute for Health Innovation & Technology (iHealthtech), Mechanobiology Institute, National University of Singapore, Singapore

The future of healthcare wearables lies in continuous sensing and monitoring in an unobtrusive manner. However, for use as medical wearables, it is important to confer flexibility and stretchability, while maintaining its sensitivity and robustness. Here, we develop novel liquid metal-based microfluidic and microtubular sensors that possess high flexibility, durability, and sensitivity. The sensors comprise a soft elastomer-based microfluidic template encapsulating a conductive liquid metal which serves as the active sensing element of the device. This sensor is capable of distinguishing and quantifying the various vital signals as well as bodily movements it is subjected to. We demonstrated healthcare applications of our sensors in rehabilitation, artificial sensing, disease tracking such as that for diabetic patients and virtual reality training as well as rehabilitation in the health metaverse. Overall, our work highlights the potential of the liquid-based microfluidic sensing platforms in a wide range of healthcare applications and further facilitates the exploration and realization of functional liquid-state device technology.

Chemically Programmable Isothermal PCR Enables Rapid Nucleic Acid-based Bioanalysis at the Point of Need
化学的にプログラム可能な等温PCRにより必要に応じて行える迅速な核酸ベースバイオ分析
Victor Ugaz, Professor & Interim Department Head, Texas A&M University, United States of America

The polymerase chain reaction (PCR) and its variants are analytical gold standards for nucleic acid-based testing that play a critical role in diagnosing and monitoring infectious diseases. But robust portable PCR-based testing at the point of need (PON) remains elusive, especially in resource-limited settings, because the required thermocycling instrumentation is neither amenable to nor validated for operation in a portable format. This presentation describes a microfluidic platform that overcomes these barriers by exploiting the ability to perform isothermal PCR via natural convection. We apply “chemical programming” to manipulate the interplay between the PCR biochemistry and the microscale convective flow field, enabling 100% repeatability to be achieved in a format that can be easily and cost-effectively manufactured, dramatically increasing simplicity, portability, and affordability. We demonstrate rapid replication of targets associated with multiple pathogens, including SARS-CoV-2, attaining performance rivaling ultra-fast PCR instruments while using lower reagent concentrations than conventional protocols. This discovery paves the way for widespread adoption of PCR-based analysis at the PON, making it possible to bring lab-quality diagnostics to decentralized settings.

Emerging Trends in Microfluidics Inspired by Nature
自然に想を得たマイクロフルイディクスにおける新たな動向
Anderson Shum, Professor, Department of Mechanical Engineering; Director, Advanced Biomedical Instrumentation Centre, University of Hong Kong, Hong Kong

The field of microfluidics has led to a wide variety of advances in bioanalyses, fluid characterization, materials design and many other applications. Microfluidics is now commonly used as a tool well integrated into the final application. Further development of the field of microfluidics requires new ideas in the design of the microfluidic devices and strategies in general. In this talk, I will discuss a few potential trends in the devices, for instance, by making the devices more responsive to the environment, enhancing the exchange of materials with the micro-environment and extending to a wider range of fluids. The resultant devices enable microfluidics to capture advantages and features that are often observed in Nature, such as in plants, vasculature, and cellular environment.

RNA Sequencing using a Nanofluidic Device with Dual In-Plane Nanopore Sensors
RNAシーケンシングを用いた2重面内型ナノポアのナノ流体デバイス
Steve Soper, Foundation Distinguished Professor, Director, Center of BioModular Multi-scale System for Precision Medicine, The University of Kansas, United States of America

With the development of next generation sequencing (NGS), the field of transcriptomics has seen tremendous advancements opening up opportunities for improved diagnostics of diseases such as cancers and infectious diseases. RNA sequencing enables measurement of single nucleotide variants (SNVs), insertions and deletions, detection of different transcript isoforms, splice variants, and chimeric gene fusions. Although NGS has been useful for unraveling RNA structure and function, several technical difficulties remain including the need for reverse transcription and PCR amplification, which can mask epitranscriptomic modifications. To address NGS issues, we are developing an exciting new sequencing technology called Exonuclease Time-of-Flight (X-ToF) for the label-free detection and identification of single molecules. The hypothesis behind X-ToF is, “individual molecules moving electrokinetically through a 2D nanotube will experience a time-of-flight (ToF) that is dependent upon its molecular identity.”  X-ToF is a nanofluidic device comprised of input/output channels, a nano-scale enzymatic bioreactor, and a flight tube equipped with a pair of in-plane nanopores to measure the ToF of a single ribonucleotide monophosphate (rNMP). Each rNMP is produced from an unamplified RNA molecule using a processive exoribonuclease, XRN1. In this presentation we will discuss the high rate manufacturing of the X-ToF chip with sub-5 nm in-plane nanopores using nano-injection molding from a cyclic olefin polymer (COP) plastic. The in-plane pores are situated at either end of a nanochannel (50 x 50 nm; 5 µm long) that generate current transient signals to detect and deduce the identity of rNMPs. The ToF is dependent on the apparent electrophoretic mobility of the molecule. The identity is determined from the ToF, the current transient amplitudes, and dwell times using multi-parameter Principle Component Analysis (PCA). We will show the ability to detect (detection efficiency ~100%) single rNMPs with identification accuracies >98%. Finally, we will discuss the generation of solid-phase nano-bioreactors using XRN1, which can cleave ssRNA in the 5’ to 3’direction releasing rNMPs. Unique properties of the immobilized enzyme will be presented in terms of its processivity and kinetic rate of cleaving single RNA molecules.

Title to be Confirmed.
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Valerie Taly, CNRS Research Director, Professor and Group leader Translational Research and Microfluidics, Universite Paris Cite, France

Title to be Confirmed.
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Amy Shen, Professor, Okinawa Institute of Science and Technology Graduate University, Japan

Title to be Confirmed.
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Lydia Sohn, Almy C. Maynard and Agnes Offield Maynard Chair in Mechanical Engineering, University of California-Berkeley, United States of America

Modular Design of 3D Printed Microfluidics for Bioprocess Applications
バイオプロセス用途向け3Dプリントマイクロフルイディクスのモジュラーデザイン
Noah Malmstadt, Professor, Mork Family Dept. of Chemical Engineering & Materials Science, University of Southern California, United States of America

As 3D printing replaces traditional clean room manufacturing for microfluidic engineering applications, it’s becoming clear that this transition offers not only lower cost and faster design iterations, but also new opportunities for fluidic routing and control that are only possible due to the inherent three-dimensional nature of these systems. Over the past several years, we have developed design principles that take advantage of this three-dimensionality, as well as demonstrating several applications that benefit from this approach. At the core of these design principles is the modularity of microfluidic unit operations. In addition to designing prototypical unit operations such as mixers, splitters, flow focusers, droplet generators, thermal and optical sensors, and world-to-chip interfaces, we have developed systematic approaches to combining these modules into microfluidic circuits with predictable behaviors. This approach can be used to rapidly prototype complex microfluidic operations by assembling physically distinct modules as well as to design monolithic microfluidic devices which can be printed in a single run. We have demonstrated the power of this approach by building several micro- and milifluidic systems for bio-analysis and bioprocess applications. These include systems for biomarker diagnostics, automated high-throughput affinity screening, and rapid manufacturing of vaccine lipid nanoparticles. We have also demonstrated how entire systems can be treated as modules, allowing for scaling of bioprocess production lines by massive parallelization.

Lab on a Particle Technology for Functional Discovery of Rare and Unconventional T-Cell Receptors
希少および非従来型T細胞受容体の機能探索のためのラボオン粒子技術
Dino Di Carlo, Armond and Elena Hairapetian Chair in Engineering and Medicine, Professor and Vice Chair of Bioengineering, University of California-Los Angeles, United States of America

Lab on a particle technologies are emerging as a rapidly adopted platform for performing single-cell functional screening leveraging standard instrumentation, such as flow cytometers and microfluidic single-cell sequencing platforms. Each cell and its secreted products can be analyzed and sorted using widely available fluorescence activating cell sorters operating at up to a 1000 cells per second, promising to democratize single-cell technologies. The platform enables sorting cells based on secreted products for the discovery of antibodies, the development of cell lines producing recombinant products, and the selection of functional cells for cell therapies. Here, I will describe our progress in using nanovial particles for the characterization and discovery of antigen-specific and metabolite-specific T cells. Nanovials enable selective binding, functional characterization of secretion of cytokines and other effector molecules, and sorting of T cells for downstream T-cell receptor sequencing and functional annotation. We apply oligo barcoding technology to both multiplex target antigens and functional performance and link this to the TCR sequence information. Nanovials can enable more rapid discovery of cancer-specific TCRs and TCRs against metabolites presented by unconventional MHC-like molecules, with high accuracy, promising to transform the discovery of critical recognition elements for improved T cell therapies.

Rapid and Signal Crowdedness-Robust In-Situ Sequencing through Hybrid Block Coding
ブロック符号化のハイブリッドによる迅速かつ信号の混雑にロバストなIn-Situシーケンシング
Yanyi Huang, Professor, Peking University, China

Spatial transcriptomics technology has revolutionized our understanding of cell types and tissue organization, opening new possibilities for researchers to explore transcript distributions at subcellular levels. However, existing methods have limitations in resolution, sensitivity, or speed. To overcome these challenges, we introduce SPRINTseq (Spatially Resolved and signal-diluted Next-generation Targeted sequencing), an innovative in situ sequencing strategy that combines hybrid block coding and molecular dilution strategies. Our method enables fast and sensitive high-resolution data acquisition, as demonstrated by recovering over 142 million transcripts from 453,843 cells from four mouse brain coronal slices in less than two days. Using this advanced technology, we uncover the cellular and subcellular molecular architecture of Alzheimer's disease, providing unprecedented insights into abnormal cellular behaviors and their subcellular mRNA distribution. This improved spatial transcriptomics technology holds great promise for exploring complex biological processes and disease mechanisms.

Analytical Assays on Paper Platforms: As Simple as Possible
ペーパープラットフォーム上の分析アッセイ：可能な限りシンプルに
Daniel Citterio, Professor, Keio University, Japan

There is a continuously growing demand for analytical assays that can be performed on-site by minimally trained operators without the need for sophisticated laboratory equipment and reagent handling. In this context, (microfluidic) paper-based analytical devices (µ)PADs, including lateral-flow immunoassays (LFIAs), have gained significant attention, particularly for point-of-care testing in medical diagnostics. Over the past few years, we have demonstrated that certain analytical assays conventionally depending on benchtop instruments can be implemented into simple paper platforms not requiring any reagent handling. Examples include paper-based devices for the naked eye “calibration-free” colorimetric semiquantitative analysis of clinically relevant parameters that can be operated by minimally trained users with minimal risk of incorrect result misinterpretation. The presentation will also introduce one of our latest efforts towards boosting assay sensitivity on paper by adapting the CRISPR/Cas system to a paper-based analytical platform in the form of a simple, rapid, and highly sensitive detection method for non-nucleic acid targets by integrating CRISPR/Cas12a and an enzyme-linked immuno-sorbent assay (ELISA).