Lab on a Chip最新文献

Despite considerable animal sacrifices and investments, drug development often falters in clinical trials due to species differences. To address this issue, specific in vitro models, such as organ-on-a-chip technology using human cells in microfluidic devices, are recognized as promising alternatives. Among the various organs, the human small intestine plays a pivotal role in drug development, particularly in the assessment of digestion and nutrient absorption. However, current intestine-on-a-chip devices struggle to accurately replicate the complex 3D tubular structures of the human small intestine, particularly when it comes to integrating a variety of cell types effectively. This limitation is primarily due to conventional fabrication methods, such as soft lithography and replica molding. In this research, we introduce a novel coaxial bioprinting method to construct 3D tubular structures that closely emulate the organization and functionality of the small intestine with multiple cell types. To ensure stable production of these small intestine-like tubular structures, we analyzed the rheological properties of bioinks to select the most suitable materials for coaxial bioprinting technology. Additionally, we conducted biological assessments to validate the gene expression patterns and functional attributes of the 3D intestine-on-a-chip. Our 3D intestine-on-a-chip, which faithfully replicates intestinal functions and organization, demonstrates clear superiority in both structure and biological function compared to the conventional 2D model. This innovative approach holds significant promise for a wide range of future applications.

Assisted reproductive technologies (ART) are pivotal for contemporary reproductive medicine and species conservation. However, the manual handling required in these processes introduces stress that can compromise oocyte and embryo quality. This study introduces OoTrap, a novel fluidic device designed to streamline ART workflows by facilitating the capture and maturation of oocytes in a compact unit. The device also reintroduces mechanical forces similar to those in the in vivo environment, which are often missing in conventional systems. OoTrap operates in both static and perfusion-based modes, offering flexibility and optimal conditions for oocyte maturation. Notably, OoTrap achieved higher in vitro maturation (IVM) rates under perfusion, produced oocytes with fewer chromosomal abnormalities, and maintained spindle morphology integrity. The incorporation of a heating system and a 3D-printed syringe pump enabled IVM outside the incubator, making OoTrap suitable for field applications. The results highlight the potential of OoTrap to enhance ART outcomes by reducing manual handling, providing a controlled microenvironment, and offering a practical solution for field-based ART applications.

Multi-organ-on-chip systems (MOOCs) have the potential to mimic communication between organ systems and reveal mechanisms of health and disease. However, many existing MOOCs are challenging for non-experts to implement due to complex tubing, electronics, or pump mechanisms. In addition, few MOOCs have incorporated immune organs such as the lymph node (LN), limiting their applicability to model critical events such as vaccination. Here we developed a 3D-printed, user-friendly device and companion tubing-free impeller pump with the capacity to co-culture two or more tissue samples, including a LN, under a recirculating common media. Native tissue structure and immune function were incorporated by maintaining slices of murine LN tissue ex vivo in 3D-printed mesh supports for at least 24 h. In a two-compartment model of a LN and an upstream injection site in mock tissue, vaccination of the multi-compartment chip was similar to in vivo vaccination in terms of locations of antigen accumulation and acute changes in activation markers and gene expression in the LN. We anticipate that in the future, this flexible platform will enable models of multi-organ immune responses throughout the body.

This work introduces a high-throughput setup for Raman analysis of various flowing fluids, both transparent and non-transparent. The setup employs a microfluidic cell, used with an external optical setup, to control the sample flow's position and dimensions via 3-dimensional hydrodynamic focusing. This approach, in contrast to the prevalent use of fused silica capillaries, reduces the risk of sample photodegradation and boosts measurement efficiency, enhancing overall system throughput. The microfluidic cell has been further evolved to laminate two distinct flows from different samples in parallel. Using line excitation, both samples can be simultaneously excited without moving parts, further increasing throughput. This setup also enables real-time monitoring of phenomena like mixing or potential reactions between the two fluids. This development could significantly advance the creation of highly sensitive, high-throughput sensors for fluid composition analysis.

Cell transfer printing plays an essential role in biomedical research and clinical diagnostics. Traditional bioadhesion-based methods often necessitate complex surface modifications and offer limited control over the quantity of transferred cells. There is a critical need for a modification-free, non-labeling, and high-throughput cell transfer printing technique. In this study, an adhesion-free cellular transfer printing method based on vibration-induced microstreaming is introduced. By adjusting the volume of the microcavity, the number of cells transferred per microtiter well can be realized to the level of a single cell. Additionally, it allows for precise control of large-scale cellular spatial distribution, leading to the formation of biomimetic patterns. Moreover, the demonstrated biocompatibility and high throughput of this cell transfer printing method highlight its potential utility. The correspondence of the transferred cell amount to the vibration and frequencies allows the system to exhibit excellent tunability of the transferred cell amount and pattern. This bioadhesion-free cell transfer printing method holds promise for advancing cell manipulation in biomedical research and analysis.

Microwell technology is crucial in biological applications due to its ability to handle small sample sizes and perform numerous assays efficiently. This study aimed to develop a novel technique for microwell fabrication using pressure-assisted steam technology, offering lower cost, simplicity, and high reproducibility. Mechanical properties of microwell surfaces were successfully controlled and characterized, making them suitable for DNA capture. The application of gold coating generated an electric field within designed microwells, facilitating stable DNA detection. These microwells exhibited effective DNA sensing capabilities, validated using fluorescently stained lambda DNA at various concentrations (86, 8.6, and 0.86 ng μL⁻¹). In particular, the 2.8 mm microwell showed a greater change in fluorescence intensity depending on DNA concentration than other microwells. At a concentration of 0.86 ng μL⁻¹, to assess producibility using relative standard deviation (RSD) values as a DNA sensor, they were measured as 5.29, 2.76, and 1.85% for 1, 1.7, and 2.8 mm microwells, respectively. These results indicated that our proposed microwell exhibited efficient performance and good reproducibility. We believe that the developed method could be potentially used for high-throughput analysis as a biosensor for DNA applications.

Many proteins, especially eukaryotic proteins, membrane proteins and protein complexes, are challenging to study because they are difficult to purify in their native state without disrupting the interactions with their partners. Hence, our lab developed a novel purification technique employing Nanobodies® (Nbs). This technique, called nanobody exchange chromatography (NANEX), utilises an immobilised low-affinity Nb to capture the target protein, which is subsequently eluted – along with its interaction partners – by introducing a high-affinity Nb. In line with the growing trend towards studying proteins in smaller sample sizes, the present study validates miniaturisation of NANEX in a packed bed microfluidic (μNANEX) chip. This μNANEX setup integrates up to five submicroliter silicon chips, enabling fully automated and reproducible purifications within minutes. Additionally, a digital twin model of the μNANEX column, which accurately predicts the effect of the reaction kinetics and mass transfer on the elution peaks, has been validated over a broad range of experimental conditions. The effectiveness of the method is demonstrated with Nbs binding to the green fluorescent protein (GFP), allowing streamlined purification of any GFP fusion protein from biological samples. Specifically, we used μNANEX to purify 0.1–1 μg of GFP-fused yeast proteins from 20 μL crude lysate and identified their interaction partners via mass spectrometry, showing that μNANEX purification preserves protein complexes.

Materials with high light-to-heat conversion efficiencies offer valuable strategies for remote heating. These materials find wide applications in photothermal therapy, water distillation, and gene delivery. In this study, we investigated a universal coating method to impart photothermal features to various surfaces. Polydopamine, a well-known adhesive material inspired by mussels, served as an intermediate layer to anchor polyethyleneimine and capture gold nanoparticles. Subsequently, the coated surface underwent electroless gold deposition to improve photothermal heating efficiency by increasing light absorption. This process was analyzed through scanning electron microscopic imaging and absorbance measurements. To demonstrate functionality, the coated surface was photothermally heated using a light-emitting diode controlled with a microprocessor, targeting the metal regulatory transcription factor 1 gene-a marker for osteoarthritis-and the S gene of the severe fever with thrombocytopenia syndrome virus. Successful amplification of the target genes was confirmed after 34 polymerase chain reaction cycles in just 12 min, verified by gel electrophoresis, demonstrating its diagnostic applicability. Overall, this simple photothermal coating method provides versatile utility, and is applicable to diverse surfaces such as membranes, tissue culture dishes, and microfluidic systems.

Microplastics (MPs) are pervasive pollutants present in various environments. They have the capability to infiltrate the human gastrointestinal tract through avenues like water and food, and ultimately accumulating within the liver. However, due to the absence of reliable platforms, the transportation, uptake, and damage of microplastics in the gut-liver axis remain unclear. Here, we present the development of a gut-liver-on-a-chip (GLOC) featuring biomimetic intestinal peristalsis and a dynamic hepatic flow environment, exploring the translocation in the intestines and accumulation in the liver of MPs following oral ingestion. In comparison to conventional co-culture platforms, this chip has the capability to mimic essential physical microenvironments found within the intestines and liver (e.g., intestinal peristalsis and liver blood flow). It effectively reproduces the physiological characteristics of the intestine and liver (e.g., intestinal barrier and liver metabolism). Moreover, we infused polyethylene MPs with a diameter of 100 nm into the intestinal and hepatic chambers (concentrations ranging from 0 to 1 mg mL^-1). We observed that as intestinal peristalsis increased (0%, 1%, 3%, 5%), the transport rate of MPs decreased, while the levels of oxidative stress and damage in hepatic cells decreased correspondingly. Our GLOC elucidates the process of MP transport in the intestine and uptake in the liver following oral ingestion. It underscores the critical role of intestinal peristalsis in protecting the liver from damage, and provides a novel research platform for assessing the organ-specific effects of MPs.

Sperm motility is a primary criterion for selecting viable and functional sperm in assisted reproduction, where the most motile sperm are used to increase the likelihood of successful conception. Traditional chemical agents to enhance motility pose embryo-toxicity risks, necessitating safer alternatives. This study investigates the use of low-intensity pulsed ultrasound exposure as a non-invasive treatment within an acoustofluidic device to maintain sperm motility. We utilized a droplet-based platform to examine the effects of repeated ultrasound pulses on single human sperm cells. Our findings demonstrate that repeated pulsed ultrasound maintains sperm motility over an hour, with significant improvements in motility parameters by at least 25% as compared to non-exposed sperm. Moreover, we show that the motility enhancements by repeated pulsed ultrasound are more significant in initially non-progressive sperm. Importantly, this method did not compromise sperm viability or DNA integrity. These results suggest a viable, sperm safe approach to enhance and maintain sperm motility, potentially improving assisted reproduction outcomes.