A portable and high-integrated 3D microfluidic chip for bacterial quantification and antibiotic susceptibility testing

On-site single-cell antibiotic susceptibility testing (sc-AST) provides unprecedented technical potential to improve the treatment of bacterial infections and study heterogeneous resistance to antibiotics. Herein, we developed a portable and high-integrated 3D polydimethylsiloxane (PDMS) chip to perform fast and on-site bacteria quantification and sc-AST. The 3D arrangement of the chambers significantly improved the integration of reaction units (~10000/cm2) and widened the dynamic range to 5 orders of magnitude. A capillary valve-based flow distributor was adopted for flow equidistribution in 64 parallel channels and uniform sample loading in as short as 2 s. The degassed PDMS enabled this device to independently dispense the sample into 3D chamber array with almost 100% efficiency. The quantification of Escherichia coli (E. coli) strains with various activity was accomplished in 0.5-2 h, shortened by 20 h in comparison to the traditional plate counting. The functionality of our platform was demonstrated with several effective antibiotics by determining minimum inhibitory concentrations at single-cell level. Furthermore, we utilized the lyophilization of test reagents and needle-mediated reagents rehydration to realize one-step on-site sc-AST. The results indicate that the proposed sc-AST platform is portable, highly sensitive, fast, accurate and user-friendly, thus it has the potential to facilitate precise therapy in time and monitor the treatment. Meanwhile, it could serve as an approach for investigating the mechanisms of heteroresistance at single-cell resolution.

Microfluidics 4: PDMS Chip Soft Lithography v2

Microfluidics materials are of various types and application-specific. PDMS is one of the most preferred and cost-effective solutions for research and low-volume manufacturing. After having the mold, PDMS replicas are generated by a technique called soft-lithography. This protocol describes the preparation of PDMS microchannels using SU8 molds, 3D Printed resin molds, and/or metal molds by the soft lithography technique, SLA printing, or CNC machining.

Portable all-in-one automated microfluidic system (PAMICON) with 3D-printed chip using novel fluid control mechanism

State-of-the-art microfluidic systems rely on relatively expensive and bulky off-chip infrastructures. The core of a system—the microfluidic chip—requires a clean room and dedicated skills to be fabricated. Thus, state-of-the-art microfluidic systems are barely accessible, especially for the do-it-yourself (DIY) community or enthusiasts. Recent emerging technology—3D-printing—has shown promise to fabricate microfluidic chips more simply, but the resulting chip is mainly hardened and single-layered and can hardly replace the state-of-the-art Polydimethylsiloxane (PDMS) chip. There exists no convenient fluidic control mechanism yet suitable for the hardened single-layered chip, and particularly, the hardened single-layered chip cannot replicate the pneumatic valve—an essential actuator for automatically controlled microfluidics. Instead, 3D-printable non-pneumatic or manually actuated valve designs are reported, but their application is limited. Here, we present a low-cost accessible all-in-one portable microfluidic system, which uses an easy-to-print single-layered 3D-printed microfluidic chip along with a novel active control mechanism for fluids to enable more applications. This active control mechanism is based on air or gas interception and can, e.g., block, direct, and transport fluid. As a demonstration, we show the system can automatically control the fluid in microfluidic chips, which we designed and printed with a consumer-grade 3D-printer. The system is comparably compact and can automatically perform user-programmed experiments. All operations can be done directly on the system with no additional host device required. This work could support the spread of low budget accessible microfluidic systems as portable, usable on-the-go devices and increase the application field of 3D-printed microfluidic devices.

Microfluidics 4: PDMS Chip Soft Lithography v2

Microfluidics materials are of various types and application-specific. PDMS is one of the most preferred and cost-effective solutions for research and low-volume manufacturing. After having the mold, PDMS replicas are generated by a technique called soft-lithography. This protocol describes the preparation of PDMS microchannels using SU8 molds, 3D Printed resin molds, and/or metal molds by the soft lithography technique, SLA printing, or CNC machining.

A Concentration Gradients Tunable Generator with Adjustable Position of the Acoustically Oscillating Bubbles

It is essential to control concentration gradients at specific locations for many biochemical experiments. This paper proposes a tunable concentration gradient generator actuated by acoustically oscillating bubbles trapped in the bubble channels using a controllable position based on the gas permeability of polydimethylsiloxane (PDMS). The gradient generator consists of a glass substrate, a PDMS chip, and a piezoelectric transducer. When the trapped bubbles are activated by acoustic waves, the solution near the gas–liquid interface is mixed. The volume of the bubbles and the position of the gas–liquid interface are regulated through the permeability of the PDMS wall. The tunable concentration gradient can be realized by changing the numbers and positions of the bubbles that enable the mixing of fluids in the main channel, and the amplitude of the applied voltage. This new device is easy to fabricate, responsive, and biocompatible, and therefore has great application prospects. In particular, it is suitable for biological research with high requirements for temporal controllability.

Dielectrophoretic Separation of Particles Using Microfluidic Chip with Composite Three-Dimensional Electrode

Integrating three-dimensional (3D) microelectrodes on microfluidic chips based on polydimethylsiloxane (PDMS) has been a challenge. This paper introduces a composite 3D electrode composed of Ag powder (particle size of 10 nm) and PDMS. Ethyl acetate is added as an auxiliary dispersant during the compounding process. A micromachining technique for processing 3D microelectrodes of any shape and size was developed to allow the electrodes to be firmly bonded to the PDMS chip. Through theoretical calculations, numerical simulations, and experimental verification, the role of the composite 3D microelectrodes in separating polystyrene particles of three different sizes via dielectrophoresis was systematically studied. This microfluidic device separated 20-, 10-, and 5-μm polystyrene particles nondestructively, efficiently, and accurately.

Effects of Pulse Interval and Dosing Flux on Cells Varying the Relative Velocity of Micro Droplets and Culture Solution

Microdroplet dosing to cell on a chip could meet the demand of narrow diffusion distance, controllable pulse dosing and less impact to cells. In this work, we studied the diffusion process of microdroplet cell pulse dosing in the three-layer sandwich structure of PDMS (polydimethylsiloxane)/PCTE (polycarbonate) microporous membrane/PDMS chip. The mathematical model is established to solve the diffusion process and the process of rhodamine transfer to micro-traps is simulated. The rhodamine mass fraction distribution, pressure field and velocity field around the microdroplet and cell surfaces are analyzed for further study of interdiffusion and convective diffusion effect. The cell pulse dosing time and drug delivery efficiency could be controlled by adjusting microdroplet and culture solution velocity without impairing cells at micro-traps. Furthermore, the accuracy and controllability of the cell dosing pulse time and maximum drug mass fraction on cell surfaces are achieved and the drug effect on cells could be analyzed more precisely especially for neuron cell dosing.