3D Printed Microfluidic Devices

3D printing has revolutionized the microfabrication prototyping workflow over the past few years. With the recent improvements in 3D printing technologies, highly complex microfluidic devices can be fabricated via single-step, rapid, and cost-effective protocols as a promising alternative to the time consuming, costly and sophisticated traditional cleanroom fabrication. Microfluidic devices have enabled a wide range of biochemical and clinical applications, such as cancer screening, micro-physiological system engineering, high-throughput drug testing, and point-of-care diagnostics. Using 3D printing fabrication technologies, alteration of the design features is significantly easier than traditional fabrication, enabling agile iterative design and facilitating rapid prototyping. This can make microfluidic technology more accessible to researchers in various fields and accelerates innovation in the field of microfluidics. Accordingly, this Special Issue seeks to showcase research papers, short communications, and review articles that focus on novel methodological developments in 3D printing and its use for various biochemical and biomedical applications

Smart-Phone Based Magnetic Levitation for Measuring Densities

Magnetic levitation, which uses a magnetic field to suspend objects in a fluid, is a powerful and versatile technology. We develop a compact magnetic levitation platform compatible with a smart-phone to separate micro-objects and estimate the density of the sample based on its levitation height. A 3D printed attachment is mechanically installed over the existing camera unit of a smart-phone. Micro-objects, which may be either spherical or irregular in shape, are suspended in a paramagnetic medium and loaded in a microcapillary tube which is then inserted between two permanent magnets. The micro-objects are levitated and confined in the microcapillary at an equilibrium height dependent on their volumetric mass densities (causing a buoyancy force toward the edge of the microcapillary) and magnetic susceptibilities (causing a magnetic force toward the center of the microcapillary) relative to the suspending medium. The smart-phone camera captures magnified images of the levitating micro-objects through an additional lens positioned between the sample and the camera lens cover. A custom-developed Android application then analyzes these images to determine the levitation height and estimate the density. Using this platform, we were able to separate microspheres with varying densities and calibrate their levitation heights to known densities to develop a technique for precise and accurate density estimation. We have also characterized the magnetic field, the optical imaging capabilities, and the thermal state over time of this platform

A continuous mapping of sleep states through association of EEG with a mesoscale cortical model

Here we show that a mathematical model of the human sleep cycle can be used to obtain a detailed description of electroencephalogram (EEG) sleep stages, and we discuss how this analysis may aid in the prediction and prevention of seizures during sleep. The association between EEG data and the cortical model is found via locally linear embedding (LLE), a method of dimensionality reduction. We first show that LLE can distinguish between traditional sleep stages when applied to EEG data. It reliably separates REM and non-REM sleep and maps the EEG data to a low-dimensional output space where the sleep state changes smoothly over time. We also incorporate the concept of strongly connected components and use this as a method of automatic outlier rejection for EEG data. Then, by using LLE on a hybrid data set containing both sleep EEG and signals generated from the mesoscale cortical model, we quantify the relationship between the data and the mathematical model. This enables us to take any sample of sleep EEG data and associate it with a position among the continuous range of sleep states provided by the model; we can thus infer a trajectory of states as the subject sleeps. Lastly, we show that this method gives consistent results for various subjects over a full night of sleep and can be done in real time

Transport phenomena and pharmacokinetics of anti-HIV microbicide drug delivery

Microbicides are agents that can be applied to vaginal mucosal surfaces with the goal of blocking the transmission of sexually transmitted pathogens including HIV. They can be formulated in various delivery vehicles, including gels, creams, lotions, tablets or films. A recent study in South Africa has confirmed, for the first time, that a vaginal gel formulation of the antiretroviral drug Tenofovir, when topically applied, significantly inhibits sexual HIV transmission to women. However the gel for this drug, and anti-HIV microbicide vehicles in general, have not been designed using an understanding of how their spreading and retention in the vagina govern successful drug delivery. Elastohydrodynamic lubrication theory can be applied to model spreading of microbicide gels and films. This should incorporate the full rheological behavior, including how rheological properties change due to contact with, and dilution by, ambient vaginal fluids. A Carreau-like non-Newtonian model is adopted as a rheological model. The effects of three important factors, i.e., dilution by vaginal fluid, a yield stress of the gel and swelling, are investigated. Dilution accelerates the coating flow by creating a slippery region near the vaginal wall akin to a dilution boundary layer. The concept of a yield stress fluid is attractive because such a material may tend to stay in place after coating vulnerable surfaces. We show that the lubrication approximation, which eases the efforts of design, may be applied to the elastic wall-squeezing problem for a prototype gel with yield stress. For the important presence of significant swelling of the gel by absorption of vaginal fluid, the model indicates more rapid coating. Then we study films that can be preferable over gels due to several reasons in many parts of the world. A model is developed for transient swelling and spreading of a film, and subsequent distribution of an active drug throughout the vaginal lumen. We investigate the effects of combination of many factors.The theory developed here improves our understanding of the biophysics of microbicide gel and film flows in vivo. Ultimately, this dissertation contributes to a methodology that provides objective evaluations of microbicide gel and film performance and in the design of new, improved vehicles

Editorial for the Special Issue on 3D Printed Microfluidic Devices

Three-dimensional (3D) printing has revolutionized the microfabrication prototyping workflow over the past few years. [...

Emerging Anti-Fouling Methods: Towards Reusability of 3D-Printed Devices for Biomedical Applications

Microfluidic devices are used in a myriad of biomedical applications such as cancer screening, drug testing, and point-of-care diagnostics. Three-dimensional (3D) printing offers a low-cost, rapid prototyping, efficient fabrication method, as compared to the costly—in terms of time, labor, and resources—traditional fabrication method of soft lithography of poly(dimethylsiloxane) (PDMS). Various 3D printing methods are applicable, including fused deposition modeling, stereolithography, and photopolymer inkjet printing. Additionally, several materials are available that have low-viscosity in their raw form and, after printing and curing, exhibit high material strength, optical transparency, and biocompatibility. These features make 3D-printed microfluidic chips ideal for biomedical applications. However, for developing devices capable of long-term use, fouling—by nonspecific protein absorption and bacterial adhesion due to the intrinsic hydrophobicity of most 3D-printed materials—presents a barrier to reusability. For this reason, there is a growing interest in anti-fouling methods and materials. Traditional and emerging approaches to anti-fouling are presented in regard to their applicability to microfluidic chips, with a particular interest in approaches compatible with 3D-printed chips

Assessing the Reusability of 3D-Printed Photopolymer Microfluidic Chips for Urine Processing

Three-dimensional (3D) printing is emerging as a method for microfluidic device fabrication boasting facile and low-cost fabrication, as compared to conventional fabrication approaches, such as photolithography, for poly(dimethylsiloxane) (PDMS) counterparts. Additionally, there is an increasing trend in the development and implementation of miniaturized and automatized devices for health monitoring. While nonspecific protein adsorption by PDMS has been studied as a limitation for reusability, the protein adsorption characteristics of 3D-printed materials have not been well-studied or characterized. With these rationales in mind, we study the reusability of 3D-printed microfluidics chips. Herein, a 3D-printed cleaning chip, consisting of inlets for the sample, cleaning solution, and air, and a universal outlet, is presented to assess the reusability of a 3D-printed microfluidic device. Bovine serum albumin (BSA) was used a representative urinary protein and phosphate-buffered solution (PBS) was chosen as the cleaning agent. Using the 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) fluorescence detection method, the protein cross-contamination between samples and the protein uptake of the cleaning chip were assessed, demonstrating a feasible 3D-printed chip design and cleaning procedure to enable reusable microfluidic devices. The performance of the 3D-printed cleaning chip for real urine sample handling was then validated using a commercial dipstick assay

Shape Fidelity of 3D-Bioprinted Biodegradable Patches

There is high demand in the medical field for rapid fabrication of biodegradable patches at low cost and high throughput for various instant applications, such as wound healing. Bioprinting is a promising technology, which makes it possible to fabricate custom biodegradable patches. However, several challenges with the physical and chemical fidelity of bioprinted patches must be solved to increase the performance of patches. Here, we presented two hybrid hydrogels made of alginate-cellulose nanocrystal (CNC) (2% w/v alginate and 4% w/v CNC) and alginate-TEMPO oxidized cellulose nanofibril (T-CNF) (4% w/v alginate and 1% w/v T-CNC) via ionic crosslinking using calcium chloride (2% w/v). These hydrogels were rheologically characterized, and printing parameters were tuned for improved shape fidelity for use with an extrusion printing head. Young’s modulus of 3D printed patches was found to be 0.2–0.45 MPa, which was between the physiological ranges of human skin. Mechanical fidelity of patches was assessed through cycling loading experiments that emulate human tissue motion. 3D bioprinted patches were exposed to a solution mimicking the body fluid to characterize the biodegradability of patches at body temperature. The biodegradation of alginate-CNC and alginate-CNF was around 90% and 50% at the end of the 30-day in vitro degradation trial, which might be sufficient time for wound healing. Finally, the biocompatibility of the hydrogels was tested by cell viability analysis using NIH/3T3 mouse fibroblast cells. This study may pave the way toward improving the performance of patches and developing new patch material with high physical and chemical fidelity for instant application

Hemp-Based Microfluidics

Hemp is a sustainable, recyclable, and high-yield annual crop that can be used to produce textiles, plastics, composites, concrete, fibers, biofuels, bionutrients, and paper. The integration of microfluidic paper-based analytical devices (µPADs) with hemp paper can improve the environmental friendliness and high-throughputness of µPADs. However, there is a lack of sufficient scientific studies exploring the functionality, pros, and cons of hemp as a substrate for µPADs. Herein, we used a desktop pen plotter and commercial markers to pattern hydrophobic barriers on hemp paper, in a single step, in order to characterize the ability of markers to form water-resistant patterns on hemp. In addition, since a higher resolution results in densely packed, cost-effective devices with a minimized need for costly reagents, we examined the smallest and thinnest water-resistant patterns plottable on hemp-based papers. Furthermore, the wicking speed and distance of fluids with different viscosities on Whatman No. 1 and hemp papers were compared. Additionally, the wettability of hemp and Whatman grade 1 paper was compared by measuring their contact angles. Besides, the effects of various channel sizes, as well as the number of branches, on the wicking distance of the channeled hemp paper was studied. The governing equations for the wicking distance on channels with laser-cut and hydrophobic side boundaries are presented and were evaluated with our experimental data, elucidating the applicability of the modified Washburn equation for modeling the wicking distance of fluids on hemp paper-based microfluidic devices. Finally, we validated hemp paper as a substrate for the detection and analysis of the potassium concentration in artificial urine

CRISPR-Cas-Integrated LAMP

Pathogen-specific point-of-care (PoC) diagnostic tests have become an important need in the fight against infectious diseases and epidemics in recent years. PoC diagnostic tests are designed with the following parameters in mind: rapidity, accuracy, sensitivity, specificity, and ease of use. Molecular techniques are the gold standard for pathogen detection due to their accuracy and specificity. There are various limitations in adapting molecular diagnostic methods to PoC diagnostic tests. Efforts to overcome limitations are focused on the development of integrated molecular diagnostics by utilizing the latest technologies available to create the most successful PoC diagnostic platforms. With this point of view, a new generation technology was developed by combining loop-mediated isothermal amplification (LAMP) technology with clustered regularly interspaced short palindromic repeat (CRISPR)-associated (CRISPR-Cas) technology. This integrated approach benefits from the properties of LAMP technology, namely its high efficiency, short turnaround time, and the lack of need for a complex device. It also makes use of the programmable function of CRISPR-Cas technology and the collateral cleavage activity of certain Cas proteins that allow for convenient reporter detection. Thus, this combined technology enables the development of PoC diagnostic tests with high sensitivity, specificity, and ease of use without the need for complicated devices. In this review, we discuss the advantages and limitations of the CRISPR/Cas combined LAMP technology. We review current limitations to convert CRISPR combined LAMP into pathogen-specific PoC platforms. Furthermore, we point out the need to design more useful PoC platforms using microfabrication technologies by developing strategies that overcome the limitations of this new technology, reduce its complexity, and reduce the risk of contamination