Low-cost Microfluidic Platform Research Articles

Development of novel, simple and low–cost microfluidic platform for supporting 3D dynamic cell culture

Cell culture models more accurately would be of significant value to the medical field and pharmaceutical industry. To achieve this goal, microfluidic cell culture platforms are created and improved for modeling the native cell microenvironment because they can precisely reconstruct in vivo cellular behavior. In this study, a 3D low-cost microfluidic device is used to compare the difference between the static and dynamic environment in 3D cell culture. Cells were seeded in the microfluidic device, and to produce the fluidic flow, the pump was used with the set speed was 0.045ml/min. In 3D cell culture, the viability of cells was monitored by size growth of the spheroids for 7 days. All systems were designed and optimized without leakage of the medium. In the results, the 3D dynamic condition showed a faster increase in size than in the static condition. Overall, the study was prepared for microfluidic platforms with low-cost and simple settings. Moreover, the usage of 3D microfluidic to mimic in vivo returned favorable results that were expected for drug testing in the future. 

Organ-on-a-Chip Platform with an Integrated Screen-Printed Electrode Array for Real-Time Monitoring Trans-Epithelial Barrier and Bubble Formation.

Cellular tight junctions play a key role in establishing a barrier between different compartments of the body by regulating the selective passage of different solutes across epithelial and endothelial tissues. Over the past decade, significant efforts have been conducted to develop more clinically relevant "organ-on-a-chip" models with integrated trans-epithelial electrical resistance (TEER) monitoring systems to help better understand the fundamental underpinnings of epithelial tissue physiology upon exposure to different substances. However, most of these platforms require the use of high-cost and time-consuming photolithography processes, which limits their scalability and practical implementation in clinical research. To address this need, we have developed a low-cost microfluidic platform with an integrated electrode array that allows continuous real-time monitoring of TEER and the risk of bubble formation in the microfluidic system by using scalable manufacturing technologies such as screen printing and laser processing. The integrated printed electrode array exhibited excellent stability (with less than ∼0.02 Ω change in resistance) even after long-term exposure to a complex culture medium. As a proof of concept, the fully integrated platform was tested with HMT3522 S1 epithelial cells to evaluate the tight barrier junction formation through TEER measurement and validated with standard immunostaining procedures for Zonula occludens-1 protein. This platform could be regarded as a stepping stone for the fabrication of disposable and low-cost organ and tissue-on-a-chip models with integrated sensors to facilitate studying the dynamic response of epithelial tissues to different substances in more physiologically relevant conditions.

Highly Efficient Capture and Quantification of the Airborne Fungal Pathogen Sclerotinia sclerotiorum Employing a Nanoelectrode-Activated Microwell Array.

In this study, we present a microdevice for the capture and quantification of Sclerotinia sclerotiorum spores, pathogenic agents of one of the most harmful infectious diseases of crops, Sclerotinia stem rot. The early prognosis of an outbreak is critical to avoid severe economic losses and can be achieved by the detection of a small number of airborne spores. However, the current lack of simple and effective methods to quantify fungal airborne pathogens has hindered the development of an accurate early warning system. We developed a device that remedies these limitations based on a microfluidic design that contains a nanothick aluminum electrode structure integrated with a picoliter well array for dielectrophoresis-driven capture of spores and on-chip quantitative detection employing impedimetric sensing. Based on experimental results, we demonstrated a highly efficient spore trapping rate of more than 90% with an effective impedimetric sensing method that allowed the spore quantification of each column in the array and achieved a sensitivity of 2%/spore at 5 kHz and 1.6%/spore at 20 kHz, enabling single spore detection. We envision that our device will contribute to the development of a low-cost microfluidic platform that could be integrated into an infectious plant disease forecasting tool for crop protection.

C-MEMS Derived Glassy Carbon Electrodes-Based Sensitive Electrochemical Biosensors

In this work, we have prepared glassy carbon electrodes using conventional carbon-microelectromechanical (C-MEMS) process. SU-8 photoresist has been used as the carbon precursor. The carbon thin film electrodes have then been tested by cyclic voltammetry measurements in the potential range of −800 mV to 800 mV vs Ag/AgCl electrode in presence of 10 mM potassium ferricyanide solution, without any functionalization. The samples have also been immobilized with anti-Hepatitis-B surface antigen (HBsAg) monoclonal antibody by physical adsorption. The presence of antibodies has been further confirmed by UV-VIS spectrophotometry. It has been observed that the current peak reduces distinctly and the fractional change in current magnitude is around 33% and 75% at the potential of 175 mV vs Ag/AgCl electrode for 10 fM and 100 fM concentration respectively. Hence, the presented study is the proof-of concept of electrochemical sensing behavior of HBsAg using functionalized glassy carbon electrodes which when integrated with low cost microfluidic platform can be suitable for point-of-care diagnostics.

A low-cost microfluidic platform coupled with light emitting diode for optogenetic analysis of neuronal response in C. elegans

Optogenetic method is widely used for dissecting the neuronal function and connectivity in a specific neural circuit, which can help understanding how the animal process information and generate behavior. The nematode C. elegans has a simple but complete nervous system, making it an attractive model to study the dynamics signals of neural circuits. However, in vivo analysis on neural circuits usually rely on the complex and expensive optical equipment to allow optogenetic stimulating the neuron while recording its activities in such a freely moving animal. Hence, in this paper we reported a portable optofluidic platform that works based on optical fiber illumination and functional imaging for worm optogenetic manipulation. A light beam from LED laser pen crossing the 3D-printed optical fiber channel is used to activate the neurons specific-expressed with light sensitive proteins ChR-2. The imaging light path is perpendicular to the stimulation light, which allows activating neuron precisely and measuring cellular signals simultaneously. By using such an easy-to-assemble device, optical stimulation of the specific neurons and detection of dynamic calcium responses of other neurons could be proceeded simultaneously. Thus, the developed microfluidic platform puts forward a simple, rapid and low-cost strategy for further neural circuits studies.

Microfluidic Chip based direct triple antibody immunoassay for monitoring patient comparative response to leukemia treatment.

We report a time and cost-efficient microfluidic chip for screening the leukemia cells having three specific antigens. In this method, the target blast cells are double sorted with immunomagnetic beads and captured by the 3rd antibody immobilized on the gold surface in a microfluidic chip. The captured blast cells in the chip were imaged using a bright-field optical microscope and images were analyzed to quantify the cells. First sorting was performed with nano size immunomagnetic beads and followed by 2nd sorting where micron size immunomagnetic beads were used. The low-cost microfluidic platform is made of PMMA and glass including micro size gold pads. The developed microfluidic platform was optimized with cultured B type lymphoblast cells and tested with the samples of leukemia patients. The 8 bone marrow samples of 4 leukemia patients on the initial diagnosis and on the 15th day after the start of the chemotherapy treatment were tested both with the developed microfluidic platform and the flow cytometry. A 99% statistical agreement between the two methods shows that the microfluidic chip is able to monitor the decrease in the number of blast cells due to the chemotherapy. The experiments with the patient samples demonstrate that the developed system can perform relative measurements and have a potential to monitor the patient response to the applied therapy and to enable personalized dose adjustment.

A low-cost microfluidic platform for rapid and instrument-free detection of whooping cough

Whooping cough also called Pertussis is a highly contagious respiratory infection that affects all age populations. Given recent pertussis outbreaks, there is an urgent need for a point-of-care (POC) device for rapid diagnosis of pertussis. Herein, we report a low-cost microfluidic POC device integrated with loop-mediated isothermal amplification (LAMP) technique for the rapid and accurate diagnosis of pertussis. The 3D-printed bioanalyzer housed not only the biochip but also an in-house-developed portable and fully battery-powered heater for rapid POC detection of pertussis, without the need of external electricity. The fluorescence-based results could be rapidly visualized in about one hour by the naked eye without the need for any additional instrumentation. In addition, a simple centrifuge-free sample preparation process was optimized for the efficient lysis of pertussis samples and successfully used for direct detection of bacteria in nasopharyngeal samples. High sensitivity, with a limit of detection (LOD) of 5 DNA copies per LAMP zone, and high specificity were demonstrated. We envision that the microfluidic POC device can be used in various venues such as medical clinics, schools, and other low-resource settings for the fast detection of pertussis.

Coupling High Throughput Microfluidics and Small-Angle X-ray Scattering to Study Protein Crystallization from Solution.

In this work, we propose the combination of small-angle X-ray scattering (SAXS) and high throughput, droplet based microfluidics as a powerful tool to investigate macromolecular interactions, directly related to protein solubility. For this purpose, a robust and low cost microfluidic platform was fabricated for achieving the mixing of proteins, crystallization reagents, and buffer in nanoliter volumes and the subsequent generation of nanodroplets by means of a two phase flow. The protein samples are compartmentalized inside droplets, each one acting as an isolated microreactor. Hence their physicochemical conditions (concentration, pH, etc.) can be finely tuned without cross-contamination, allowing the screening of a huge number of saturation conditions with a small amount of biological material. The droplet flow is synchronized with synchrotron radiation SAXS measurements to probe protein interactions while minimizing radiation damage. To this end, the experimental setup was tested with rasburicase (known to be very sensitive to denaturation), proving the structural stability of the protein in the droplets and the absence of radiation damage. Subsequently weak interaction variations as a function of protein saturation was studied for the model protein lysozime. The second virial coefficients (A2) were determined from the X-ray structure factors extrapolated to the origin. A2 obtained values were found to be in good agreement with data previously reported in literature but using only a few milligrams of protein. The experimental results presented here highlight the interest and convenience of using this methodology as a promising and potential candidate for studying protein interactions for the construction of phase diagrams.

Integrated, DC voltage-driven nucleic acid diagnostic platform for real sample analysis: Detection of oral cancer

We present an integrated and low-cost microfluidic platform capable of extraction of nucleic acids from real biological samples. We demonstrate the application of this platform in pathogen detection and cancer screening. The integrated platform consists of three units including a pretreatment unit for separation of nucleic acids from lysates, a preconcentration unit for concentration of isolated nucleic acids and a sensing unit localized at a designated position on the chip for specific detection of the target nucleic acid. The platform is based on various electrokinetic phenomena exhibited by ion exchange membranes in a DC electrical field that allow them to serve as molecular filters, analyte preconcentrators and sensors. In this manuscript, we describe each unit of the integrated chip separately and show specific detection of a microRNA (miRNA 146a) biomarker associated with oral cancer as a proof-of-concept experiment. This platform technology can easily be extended to other targets of interest by optimizing the properties of the ion exchange membranes and the specific probes functionalized onto the sensors.

Wound healing assay in a low-cost microfluidic platform

Cell migration is a very important stage in different pathological processes such as wound healing, vascularisation, immune response, tissue regeneration and cancer. Traditional wound healing assays are done by scratching a monolayer on standard culture surfaces. However this method can lead to data misinterpretation and poor comparability in results from different experiments. The microfluidic wound healing assay is a mature alternative to the traditional method, but it is expensive and bulky due to the need of syringe pumps and can be difficult to implement by the end user. Here we show an alternative low-cost microfluidic platform based on the MainSTREAM platform. We also demonstrate the feasibility of a microperistaltic pump to carry out these kind of assays. A laminar flow of PBS-Trypsin-PBS confined in a microfluidic channel with a previously cultured monolayer of IIB-Mel LES melanoma cells was used to generate the wound. Cell migration rate could be quantified successfully.

Microphysiological systems and low-cost microfluidic platform with analytics

A multiorgan, functional, human in vitro assay system or 'Body-on-a-Chip' would be of tremendous benefit to the drug discovery and toxicology industries, as well as providing a more biologically accurate model for the study of disease as well as applied and basic biological research. Here, we describe the advances our team has made towards this goal, as well as the most pertinent issues facing further development of these systems. Description is given of individual organ models with appropriate cellular functionality, and our efforts to produce human iterations of each using primary and stem cell sources for eventual incorporation into this system. Advancement of the 'Body-on-a-Chip' field is predicated on the availability of abundant sources of human cells, capable of full differentiation and maturation to adult phenotypes, for which researchers are largely dependent on stem cells. Although this level of maturation is not yet achievable in all cell types, the work of our group highlights the high level of functionality that can be achieved using current technology, for a wide variety of cell types. As availability of functional human cell types for in vitro culture increases, the potential to produce a multiorgan in vitro system capable of accurately reproducing acute and chronic human responses to chemical and pathological challenge in real time will also increase.