Droplet digital molecular beacon-LAMP assay via pico-injection for ultrasensitive detection of pathogens.

A pico-injection-aided digital droplet detection platform is presented that integrates loop-mediated isothermal amplification (LAMP) with molecular beacons (MBs) for the ultrasensitive and quantitative identification of pathogens, leveraging the sequence-specific detection capabilities of MBs. The microfluidic device contained three distinct functional units including droplet generation, pico-injection, and droplet counting. Utilizing a pico-injector, MBs are introduced into each droplet to specifically identify LAMP amplification products, thereby overcoming issues related to temperature incompatibility.

Automatic ultrasensitive lateral flow immunoassay based on a color-enhanced signal amplification strategy.

Lateral flow immunoassays (LFIAs) are an essential and widely used point-of-care test for medical diagnoses. However, commercial LFIAs still have low sensitivity and specificity. Therefore, we developed an automatic ultrasensitive dual-color enhanced LFIA (DCE-LFIA) by applying an enzyme-induced tyramide signal amplification method to a double-antibody sandwich LFIA for antigen detection.

Ultrasensitive Electrochemical Immunosensor for Multiplex Sandwich Bioassaying Based on the Functional Antibodies.

In vitro diagnostics (IVDs) based on electrochemical immunosensors are crucial for disease screening, diagnosis, prognosis, and treatment monitoring. However, label-free electrochemical immunosensors commonly suffer from poor specificity, leading to false positives. To address this issue, we propose a highly sensitive and precise electrochemical immunosensor for protein marker detection.

Discovery of Trametinib as an orchestrator for cytoskeletal vimentin remodeling.

The dynamic remodeling of the cytoskeletal network of vimentin intermediate filaments supports various cellular functions, including cell morphology, elasticity, migration, organelle localization, and resistance against mechanical or pathological stress. Currently available chemicals targeting vimentin predominantly induce network reorganization and shrinkage around the nucleus. Effective tools for long-term manipulation of vimentin network dispersion in living cells are still lacking, limiting in-depth studies on vimentin function and potential therapeutic applications.

Rapid droplet-based mixing for single-molecule spectroscopy.

Probing non-equilibrium dynamics with single-molecule spectroscopy is important for dissecting biomolecular mechanisms. However, existing microfluidic rapid-mixing systems for this purpose are incompatible with surface-adhesive biomolecules, exhibit undesirable flow dispersion and are often demanding to fabricate. Here we introduce droplet-based microfluidic mixing for single-molecule spectroscopy to overcome these limitations in a wide range of applications.

Towards an active droplet-based microfluidic platform for programmable fluid handling.

Droplet-based microfluidic systems have emerged as powerful alternatives to conventional high throughput screening platforms, due to their operational flexibility, high-throughput nature and ability to efficiently process small fluid volumes. However, the challenges associated with performing bespoke operations on user-defined droplets often limit their utility in screening applications that involve complex workflows. To this end, the marriage of droplet- and valve-based microfluidic technologies offers the prospect of balancing the controllability of droplet manipulations and analytical throughput.

Microfluid Switching-Induced Transient Refractive Interface.

The laminar flow interface (LFI) developed at low Reynolds numbers is one of the most prominent features of microscale flows and has been employed in a diverse range of optofluidic applications. The formation of LFIs usually requires the manipulation of multiple streams within a microchannel using a complex hydrodynamic pumping system. Herein, we present a new type of LFI that is generated by fluid switching within a three-dimensional (3D) microlens-incorporating microfluidic chip (3D-MIMC).

PDMS-PUA bi-directional replication technology and its applications.

Polydimethylsiloxane (PDMS) and polyurethane acrylate (PUA) are excellent pattern transfer materials. In this study, PDMS-PUA bi-directional replication technology is explored using the PDMS grating as a template, and relevant technical issues are discussed in detail. Special surface treatment and process optimization are applied to solve the problems of demolding, PDMS polymerization inhibition, and substrate flatness.

DropCRISPR: A LAMP-Cas12a based digital method for ultrasensitive detection of nucleic acid.

Since their discovery, CRISPR/Cas systems have been extensively exploited in nucleic acid biosensing. However, the vast majority of contemporary platforms offer only qualitative detection of nucleic acid, and fail to realize ultrasensitive quantitative detection. Herein, we report a digital droplet-based platform (DropCRISPR), which combines loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a to realize ultrasensitive and quantitative detection of nucleic acids.

Multi-compartment supracapsules made from nano-containers towards programmable release.

The assembly of nanomaterials into suprastructures offers the possibility to fabricate larger scale functional materials, whose inner structure strongly influences their functionality for a vast range of applications. In spite of the many current strategies, achieving multi-compartment structures in a targeted and versatile way remains highly challenging. Here, we describe a controllable and straightforward route to create uniform suprastructured materials with a multi-compartmentalized architecture by confining primary nanocapsules into droplets using a cross-junction microfluidic device.

Replicating the Cynandra opis Butterfly's Structural Color for Bioinspired Bigrating Color Filters.

Multilayer grating structures, such as those found on the wings of the butterfly Cynandra opis, are able to interact with light to generate structural coloration. When illuminated and viewed at defined angles, such structural color is characterized by exceptional purity and brightness. To provide further insight into the mechanism of structural coloration, two-photon laser lithography is used to fabricate bioinspired bigrating nanostructures, whose optical properties may be controlled by variation of the height and period of the grating features.

Biomimetic Water-Repelling Surfaces with Robustly Flexible Structures.

Biomimetic liquid-repelling surfaces have been the subject of considerable scientific research and technological application. To design such surfaces, a flexibility-based oscillation strategy has been shown to resolve the problem of liquid-surface positioning encountered by the previous, rigidity-based asymmetry strategy; however, its usage is limited by weak mechanical robustness and confined repellency enhancement. Here, we design a flexible surface comprising mesoscale heads and microscale spring sets, in analogy to the mushroomlike geometry discovered on springtail cuticles, and then realize this through three-dimensional projection microstereolithography.

Flexibility-Patterned Liquid-Repelling Surfaces.

Droplets impacting solid surfaces is ubiquitous in nature and of practical importance in numerous industrial applications. For liquid-repelling applications, rigidity-based asymmetric redistribution and flexibility-based structural oscillation strategies have been proven on artificial surfaces; however, these are limited by strict impacting positioning. Here, we show that the gap between these two strategies can be bridged by a flexibility-patterned design similar to a trampoline park.

Optical-Switch-Enabled Microfluidics for Sensitive Multichannel Colorimetric Analysis.

The implementation of colorimetric analysis within microfluidic environments engenders significant benefits with respect to reduced sample and reagent consumption, system miniaturization, and real-time measurement of flowing samples. That said, conventional approaches to colorimetric analysis within microfluidic channels are hampered by short optical pathlengths and single-channel configurations, which lead to poor detection sensitivities and low analytical throughputs. Although the use of multiplexed light source/photodetector modules allows for multichannel analysis, such configurations significantly increase both instrument complexity and cost.

In Situ Visualization of the Structural Evolution and Alignment of Lyotropic Liquid Crystals in Confined Flow.

Self-assembled materials such as lyotropic liquid crystals offer a wide variety of structures and applications by tuning the composition. Understanding materials behavior under flow and the induced alignment is wanted in order to tailor structure related properties. A method to visualize the structure and anisotropy of ordered systems in situ under dynamic conditions is presented where flow-induced nanostructural alignment in microfluidic channels is observed by scanning small angle X-ray scattering in hexagonal and lamellar self-assembled phases.

Laminar Flow-Based Fiber Fabrication and Encoding via Two-Photon Lithography.

In recent years, flow photolithography (FL) has emerged as a powerful synthetic tool for the creation of barcoded microparticles with complex morphologies and chemical compositions which have been shown to be useful in a range of multiplexed bioassay applications. More specifically, FL has been highly successful in producing micron-sized, encoded particles of bespoke shape, size, and color. That said, to date, FL has been restricted to generating barcoded microparticles and has lacked the ability to produce hybrid fibers which are structurally and spectrally encoded.

Liquid repellency enhancement through flexible microstructures.

Artificial liquid-repellent surfaces have attracted substantial scientific and industrial attention with a focus on creating functional topological features; however, the role of the underlying structures has been overlooked. Recent developments in micro-nanofabrication allow us now to construct a skin-muscle type system combining interfacial liquid repellence atop a mechanically functional structure. Specifically, we design surfaces comprising bioinspired, mushroom-like repelling heads and spring-like flexible supports, which are realized by three-dimensional direct laser lithography.

Self-aligned 3D microlenses in a chip fabricated with two-photon stereolithography for highly sensitive absorbance measurement.

Absorbance measurement is a widely used method to quantify the concentration of an analyte. The integration of absorbance analysis in microfluidic chips could significantly reduce the sample consumption and contribute to the system miniaturization. However, the sensitivity and limit of detection (LoD) of analysis in microfluidic chips with conventional configuration need improvements due to the limited optical pathway and unregulated light propagation.

A Counter Propagating Lens-Mirror System for Ultrahigh Throughput Single Droplet Detection.

Fluorescence-based detection schemes provide for multiparameter analysis in a broad range of applications in the chemical and biological sciences. Toward the realization of fully portable analysis systems, microfluidic devices integrating diverse functional components have been implemented in a range of out-of-lab environments. That said, there still exits an unmet and recognized need for miniaturized, low-cost, and sensitive optical detection systems, which provide not only for efficient molecular excitation, but also enhanced photon collection capabilities.

Growing and Shaping Metal-Organic Framework Single Crystals at the Millimeter Scale.

Controlling and understanding the mechanisms that harness crystallization processes is of utmost importance in contemporary materials science and, in particular, in the realm of reticular solids where it still remains a great challenge. In this work, we show that environments mimicking microgravity conditions can harness the size and shape of functional biogenic crystals such as peptide-based metal-organic frameworks (MOFs). In particular, we demonstrate formation of the largest single crystals with controlled nonequilibrium shapes of peptide-based MOFs reported to date (e.

Self-Compensating Liquid-Repellent Surfaces with Stratified Morphology.

Artificial liquid-repellent surfaces have recently attracted vast scientific attention; however, achieving mechanical robustness remains a formidable challenge before industrialization can be realized. To this end, inspired by plateaus in geological landscapes, a self-compensating strategy is developed to pave the way for the synthesis of durable repellent surfaces. This self-compensating surface comprises tall hydrophobic structural elements, which can repel liquid droplets.

Oscillatory Viscoelastic Microfluidics for Efficient Focusing and Separation of Nanoscale Species.

The ability to precisely control particle migration within microfluidic systems is essential for focusing, separating, counting, and detecting a wide range of biological species. To date, viscoelastic microfluidic systems have primarily been applied to the focusing, separation, and isolation of micrometer-sized species, with their use in nanoparticle manipulations being underdeveloped and underexplored, due to issues related to nanoparticle diffusivity and a need for extended channel lengths. To overcome such issues, we herein present sheathless oscillatory viscoelastic microfluidics as a method for focusing and separating both micrometer and sub-micrometer species.

Three-Dimensional Printed Surfaces Inspired by Bi-Gaussian Stratified Plateaus.

Wettability of artificial surfaces is attracting increasing attention for its relevant technological applications. Functional performance is often achieved by mimicking the topographical structures found in natural flora and fauna; however, surface attributes inspired by geological landscapes have so far escaped attention. We reproduced a stratified morphology of plateaus with a bi-Gaussian height distribution using a three-dimensional direct laser lithography.

An optofluidic system with integrated microlens arrays for parallel imaging flow cytometry.

In recent years, high-speed imaging has become increasingly effective for the rapid analysis of single cells in flowing environments. Single cell imaging methods typically incorporate a minimum magnification of 10× when extracting sizing and morphological information. Although information content may be significantly enhanced by increasing magnification, this is accompanied by a corresponding reduction in field of view, and thus a decrease in the number of cells assayed per unit time.

A sample-in-digital-answer-out system for rapid detection and quantitation of infectious pathogens in bodily fluids.

A variety of automated sample-in-answer-out systems for in vitro molecular diagnostics have been presented and even commercialized. Although efficient in operation, they are incapable of quantifying targets, since quantitation based on analog analytical methods (via standard curve analysis) is complex, expensive, and challenging. To address this issue, herein, we describe an integrated sample-in-digital-answer-out (SIDAO) diagnostic system incorporating DNA extraction and digital recombinase polymerase amplification, which enables rapid and quantitative nucleic acid analysis from bodily fluids within a disposable cartridge.