Microfluidic Applications of Artificial Cilia: Recent Progress, Demonstration, and Future Perspectives.

Artificial cilia-based microfluidics is a promising alternative in lab-on-a-chip applications which provides an efficient way to manipulate fluid flow in a microfluidic environment with high precision. Additionally, it can induce favorable local flows toward practical biomedical applications. The endowment of artificial cilia with their anatomy and capabilities such as mixing, pumping, transporting, and sensing lead to advance next-generation applications including precision medicine, digital nanofluidics, and lab-on-chip systems. This review summarizes the importance and significance of the artificial cilia, delineates the recent progress in artificial cilia-based microfluidics toward microfluidic application, and provides future perspectives. The presented knowledge and insights are envisaged to pave the way for innovative advances for the research communities in miniaturization.

Electrochemical biosensor with electrokinetics-assisted molecular trapping for enhancing C-reactive protein detection.

C-Reactive protein (CRP) is an essential biomarker relevant to various disease prognoses. Current biosensors require a significant amount of time for detecting CRP. To address this issue, this work proposes electrokinetic flow-assisted molecule trapping integrated with an impedance biosensor, where a driving signal in terms of a gated sine wave is provided to circularly arranged electrodes which detect proteins. To verify the biosensor's efficacy, protein aggregation on the electrode surface was evaluated through a fluorescence analysis and measurement of the electrochemical impedance spectrum (EIS). The fluorescence analysis with avidin showed that target samples largely accumulated on the electrode surface upon provision of the driving signal. The EIS measurement of CRP accumulation on the electrode surface further confirmed a significant electrokinetic phenomenon at the electrode/electrolyte interface. Even at the low CRP concentration of 10 pg/ml, the proposed device's sensitivity and reliability were as high as 3.92 pg/ml with a signal-to noise ratio (SNR) of ≥3, respectively. In addition, the protein detection time (without considering the preparation time) was minimized to as low as 90 s with the proposed device. This device's advantage is its minimal time consumption, and simple drop-analysis process flow; hence, it was used for monitoring clinical serum samples.

Shape-programmable artificial cilia for microfluidics.

The artificial ciliary motion has been known not to be hydrodynamically optimal, limiting their associated applications in the microscale flow domain. One of the major hurdles of contemporary artificial cilia is its structural rigidity, which restricts their flexibility. To address this issue, this work proposed a shape-programmable artificial cilia design with distinctive polydimethylsiloxane (PDMS) and magnetic segments distributed throughout the structure, which provided precise control for time-spatial modulation of the whole artificial cilia structure under external magnetic actuation. For the fabrication of the proposed multi-segment artificial cilia, a facile microfabrication process with stepwise mold blocking followed by the PDMS and magnetic composite casting was adopted. The hydrodynamic analysis further elucidated that the proposed artificial cilia beating induced significant flow disturbance within the flow field, and the associated application was demonstrated through an efficient mixing operation.

Infection Control Improvement of a Negative-Pressurized Pediatric Intensive Care Unit.

The COVID-19 pandemic caused by the novel SARS-CoV-2 virus raises alarming concern around the healthcare facilities due to the significant increase in patient inflow. Negative-pressurized isolation rooms have been utilized in various health care facilities to isolate the patients from active community contact. Several studies have highlighted isolation rooms improvement. However, limited knowledge is available regarding the isolation room facilities for pediatric intensive care units (PICU) to accommodate more than one pediatric patient. In this aspect, this study investigates a negative-pressurized isolation facility in PICU with minimal design modifications with the possibility that it can accommodate more than one pediatric patient. The field measurement tests were conducted to ensure the design compliance of Taiwan CDC. Then, computational fluid dynamics (CFD) was further utilized to numerically evaluate the HVAC system role and the ventilation performance towards infection control. A protected air-jet curtain system with a new ventilation layout was proposed through this study to enhance the protection for both pediatric patients and medical staff. The concentration decay was monitored and recorded within 900 s to evaluate the performance. The concentration can be reduced to 504 ppm for case 1, 620 ppm for case 2, 501 ppm for case 3, and 486 ppm for case 4. In addition, the injected bioaerosol particles could be well diluted dealing with two patients presents a good performance. The results revealed that this proposed configuration could feasibly accommodate two patients with a significant contamination control to protect the medical staff and patients.

Edible additive effects on zebrafish cardiovascular functionality with hydrodynamic assessment.

Food coloring is often used as a coloring agent in foods, medicines and cosmetics, and it was reported to have certain carcinogenic and mutagenic effects in living organisms. Investigation of physiological parameters using zebrafish is a promising methodology to understand disease biology and drug toxicity for various drug discovery on humans. Zebrafish (Danio rerio) is a well-acknowledged model organism with combining assets such as body transparency, small size, low cost of cultivation, and high genetic homology with humans and is used as a specimen tool for the in-vivo throughput screening approach. In addition, recent advances in microfluidics show a promising alternative for zebrafish manipulation in terms of drug administration and extensive imaging capability. This pilot work highlighted the design and development of a microfluidic detection platform for zebrafish larvae through investigating the effects of food coloring on cardiovascular functionality and pectoral fin swing ability. The zebrafish embryos were exposed to the Cochineal Red and Brilliant Blue FCF pigment solution in a concentration of (0.02‰, 0.2‰) cultured in the laboratory from the embryo stage to hatching and development until 9 days post fertilization (d.p.f.). In addition, zebrafish swimming behaviors in terms of pectoral fin beating towards the toxicity screening were further studied by visualizing the induced flow field. It was evidenced that Cochineal Red pigment at a concentration of 0.2‰ not only significantly affected the zebrafish pectoral fin swing behavior, but also significantly increased the heart rate of juvenile fish. The higher concentration of Brilliant Blue FCF pigment (0.2%) increased heart rate during early embryonic stages of zebrafish. However, zebrafish exposed to food coloring did not show any significant changes in cardiac output. The applications of this proposed platform can be further extended towards observing the neurobiological/hydrodynamic behaviors of zebrafish larvae for practical applications in drug tests.

Microfluidic Transportation Control of Larval Zebrafish through Optomotor Regulations under a Pressure-Driven Flow.

To perform zebrafish larvae-related experiments within a microfluidic environment, the larvae need to be anesthetized and subsequently transported into respective test sections through mechanical or manual means. However, anesthetization tends to affect larval sensory perceptions, hindering their natural behaviors. Taking into account that juvenile larvae move naturally within their environment by accessing visual as well as hydromechanical cues, this work proposes an experimental framework to transport nonanesthetized larvae within a microfluidic environment by harmonically tuning both of the aforementioned cues. To provide visual cues, computer-animated moving gratings were provided through an in-house-developed control interface that drove the larval optomotor response. In the meantime, to provide hydromechanical cues, the flow rate was tuned using a syringe pump that affected the zebrafish larvae's lateral line movement. The results obtained (corresponding to different test conditions) suggest that the magnitude of both modalities plays a crucial role in larval transportation and orientation control. For instance, with a flow rate tuning of 0.1 mL/min along with grating parameters of 1 Hz temporal frequency, the average transportation time for larvae that were 5 days postfertilization was recorded at 1.29 ± 0.49 s, which was approximately three times faster than the transportation time required only in the presence of hydromechanical cues.

Microfluidic retention of progressively motile zebrafish sperms.

Genetic manipulation of zebrafish results in thousands of mutant strains and to efficiently preserve them for future use, zebrafish sperms have been cryopreserved in various cryopreservation centers. However, cryopreservation protocols are known to alter genetic entities. Therefore, there is an urgent need for an efficient method that can select morphologically superior and progressively motile zebrafish sperms after their activation for in vitro fertilization success. However, unlike those of other mammalian species, fish sperms do not take any physical or chemical cues to travel towards the egg. Their inertness towards any external cues makes the control of their orientation in a microfluidic environment difficult. In this aspect, a new microfluidic concept was demonstrated where PDMS baffles were inserted in the sidewalls to form microscale confinement creating a flow stagnation zone towards sperm retention. Two distinct microfluidic device designs were selected to evidence the improvement in sperm retention through the unique hydrodynamic feature provided by the microchannel design. Under similar flow conditions, 44% improvement was noticed for the device with a modified baffle design in terms of sperm retrieving efficiency. It was further noticed that with a flow tuning of 0.7 µL min-1, 80% of the total sperms swimming into the retention zones was retained within a specific time window. The present work further explains the significance of the hydrodynamic dependency of zebrafish sperm kinematics that paves the way for highly efficient spermatozoan manipulation.

A Shape Memory Alloy-Based Miniaturized Actuator for Catheter Interventions.

In the current scenario of endovascular intervention, surgeons have to manually navigate the catheter within the complex vasculature of the human body under the guidance of X-ray. This manual intervention upsurges the possibilities of vessel damage due to frequent contact between the catheter and vasculature wall. In this context, a shape memory alloy-based miniaturized actuator was proposed in this study with a specific aim to reduce vessel wall related damage by improving the bending motions of the guidewire tip in a semi-automatic fashion. The miniaturized actuator was integrated with a FDA-approved guidewire and tested within a patient-specific vascular network model to realize its feasibility in the real surgical environment. The results illustrate that the miniaturized actuator gives a bending angle over 23° and lateral displacement over 900 µm to the guide wire tip by which the guidewire can be navigated with precision and possible vessel damage during the catheter intervention can certainly be minimized. In addition to it, the dynamic responses of the presented actuator were further investigated through numerical simulation in conjunction with the analytic analysis.

Sperm activation through orbital and self-axis revolutions using an artificial cilia embedded serpentine microfluidic platform.

The zebrafish sperm activation profoundly depends upon the homogeneous mixing of the sperm cells with its diluent in a quick succession as it alters the cell's extracellular medium and initiates their motility. Manual stirring, the traditional method for zebrafish sperm activation is tedious, time-consuming, and has a poor outcome. In this aspect, an artificial cilia embedded serpentine microfluidic is designed through which hydrodynamic factors of the microfluidic environment can be precisely regulated to harness uniform mixing, hence ensuring a superior sperm activation. To quantify the sperm motility, computer assisted sperm analysis software (CASA) was used whereas to quantify the generated flow field, micro particle image velocimetry (µPIV) was used. With this proposed microfluidic, 74.4% of the zebrafish sperm were activated which is 20% higher than its currently existing manual measurements. The µPIV analysis demonstrates that the curvature of the microchannel induces an orbital rotation to the flow field along the length of the microchannel together with the artificial cilia actuation which instigates a local rotation to the flow field of the artificial cilia location. The collective rotation in the whole flow field induce vorticity that promotes the change in temporal dynamics of the sperm cells towards their activation.

A noninvasive light driven technique integrated microfluidics for zebrafish larvae transportation.

Transferring the zebrafish larvae on an imaging platform has long been performed manually by the use of forceps or through mechanical pumping. These methods induce detrimental damages to the fragile bodies of zebrafish larvae during the transportation. To address this issue, in this work we are devising a light driven technique to transport zebrafish larvae within a microfluidic environment. In particular, an optomotor behavioral response of the zebrafish larvae was controlled through the computer animated moving gratings for their transportation within a microfluidics chamber. It was observed that with an optimum grating frequency of 1.5 Hz and a grating width ratio of 1:1, a 5 days-post fertilization zebrafish larva can be transported within minimum and maximum time periods of 0.63 and 2.49 s, respectively. This proposed technique can be utilized towards multi-automatic transportation of zebrafish larvae within the microfluidic environment as well as the zebrafish core facility.

An Integrated Artificial Cilia Based Microfluidic Device for Micropumping and Micromixing Applications.

A multi-purpose microfluidic device that can be used for both micromixing and micropropulsion operations has always been in demand, as it would simplify the various process flows associated with the current micro-total analysis systems. In this aspect, we propose a biomimetic artificial cilia-based microfluidic device that can efficiently facilitate both mixing and propulsion sequentially at the micro-scale. A rectangular microfluidic device consists of four straight microchannels that were fabricated using the microfabrication technique. An array of artificial cilia was embedded within one of the channel's confinement through the aforementioned technique. A series of image processing and micro-particle image velocimetry technologies were employed to elucidate the micromixing and micropropulsion phenomena. Experiment results demonstrate that, with this proposed microfluidic device, a maximum micromixing efficiency and flow rate of 0.84 and 0.089 µL/min, respectively, can be achieved. In addition to its primary application as a targeted drug delivery system, where a drug needs to be homogeneously mixed with its carrier prior to its administration into the target body, this microfluidic device can be used as a micro-total analysis system for the handling of other biological specimens.

Manipulation of zebrafish's orientation using artificial cilia in a microchannel with actively adaptive wall design.

The zebrafish is a powerful genetic model organism especially in the biomedical chapter for new drug discovery and development. The genetic toolbox which this vertebrate possesses opens a new window to investigate the etiology of human diseases with a high degree genetic similarity. Still, the requirements of laborious and time-consuming of contemporary zebrafish processing assays limit the procedure in carrying out such genetic screen at high throughput. Here, a zebrafish control scheme was initiated which includes the design and validation of a microfluidic platform to significantly increase the throughput and performance of zebrafish larvae manipulation using the concept of artificial cilia actuation. A moving wall design was integrated into this microfluidic platform first time in literature to accommodate zebrafish inside the microchannel from 1 day post-fertilization (dpf) to 6 dpf and can be further extended to 9 dpf for axial orientation control in a rotational range between 0 to 25 degrees at the minimum step of 2-degree increment in a stepwise manner. This moving wall feature was performed through the deflection of shape memory alloy wire embedded inside the microchannel controlled by the electrical waveforms with high accuracy.

Endoleak Assessment Using Computational Fluid Dynamics and Image Processing Methods in Stented Abdominal Aortic Aneurysm Models.

Endovascular aortic aneurysm repair (EVAR) is a predominant surgical procedure to reduce the risk of aneurysm rupture in abdominal aortic aneurysm (AAA) patients. Endoleak formation, which eventually requires additional surgical reoperation, is a major EVAR complication. Understanding the etiology and evolution of endoleak from the hemodynamic perspective is crucial to advancing the current posttreatments for AAA patients who underwent EVAR. Therefore, a comprehensive flow assessment was performed to investigate the relationship between endoleak and its surrounding pathological flow fields through computational fluid dynamics and image processing. Six patient-specific models were reconstructed, and the associated hemodynamics in these models was quantified three-dimensionally to calculate wall stress. To provide a high degree of clinical relevance, the mechanical stress distribution calculated from the models was compared with the endoleak positions identified from the computed tomography images of patients through a series of imaging processing methods. An endoleak possibly forms in a location with high local wall stress. An improved stent graft (SG) structure is conceived accordingly by increasing the mechanical strength of the SG at peak wall stress locations. The presented analytical paradigm, as well as numerical analysis using patient-specific models, may be extended to other common human cardiovascular surgeries.