3D-printed Cartridge Research Articles

Micro-macro SERS strategy for highly sensitive paper cartridge with trace-level molecular detection

Surface-enhanced Raman Scattering (SERS) has become a powerful spectroscopic technology for highly sensitive detection. However, SERS is still limited in the lab because it either requires complicated preparation or is limited to specific compounds, causing poor applicability for practical applications. Herein, a micro-macro SERS strategy, synergizing polymer-assisted printed process with paper-tip enrichment process, is proposed to fabricate highly sensitive paper cartridges for sensitive practical applications. The polymer-assisted printed process finely aggregates nanoparticles with a discrete degree of 1.77, and SERS results are matched with theoretical enhancement, indicating small cluster-dominated hotspots at the micro-scale and thus 41-fold SERS increase compared to other aggregation methods. The paper-tip enrichment process moves molecules in a fluid into small tips filled with plasmonic clusters, and molecular localization at hotspots is achieved by the simulation and optimization of fluidic velocity at the macro-scale, generating a 39.5-fold SERS sensibility increase in comparison with other flow methods. A highly sensitive paper cartridge contains a paper-tip and a 3D-printed cartridge, which is simple, easy-to-operate, and costs around 2 US dollars. With a detection limit of 10 −12 M for probe molecules, the application of real samples and multiple analytes achieves single-molecule level sensitivity and reliable repeatability with a 30-min standardized procedure. The micro-macro SERS strategy demonstrates its potential in practical applications that require point-of-care detection.

A smartphone-controllable molecular diagnostic platform for SARS-CoV-2 detection by reverse-transcription recombinase polymerase amplification

We constructed a portable molecular diagnostic microsystem, enabling the whole processes of genetic analysis for SARS-CoV-2. The proposed platform was a miniaturized size and was comprised of three main parts: a microfluidic device with a 3D-printed solution loading cartridge, an operating system including a tailor-made heater board, a syringe pump and an optical fluorescence detector, and a smartphone with a customized app for the remote and automatic control. The microfluidic device was designed to control the complicated fluidic flow through only the passive valves, and could perform the RNA extraction from the viral lysate via glass membranes and the target gene amplification by reverse-transcription recombinase polymerase amplification (RT-RPA). The 3D-printed cartridge allowed the facile chemical solution storage and loaded a designated solution into the microfluidic device. A smartphone with a browser-based app could control the on-and-off status of solenoid valves, the workflow of a syringe pump to automatically introduce solutions on a chip, the heater board to supply the constant temperature for RT-RPA, and the periodic activation of the Pi camera and a 488 nm laser to measure the fluorescence signal in the reaction chambers that showed the amplification profiles on the smartphone display in real-time. Using the proposed smartphone-controllable diagnostic platform, the N gene and S gene of SARS-CoV-2 were successfully detected at a limit-of-detection (LOD) of 10 copies/μL within 1 hr in a sample-to-answer-out format. Ten out of 22 clinical samples were successfully analyzed for SARS-CoV-2, demonstrating the practical applications for the on-site diagnostics of patient samples.

Mobile mini-fluorimeter for antibiotic aptasensing based on surface-plasmonic effect of burlike nanogolds enhanced by digitized imaging diagnosis

Antibiotic abuse now poses a grave threat to global ecology and bestirs public concerns about the residue issue in daily necessities. The traceability measurements along supply chain or logistic circulation have become increasingly essential given the labile nature of diverse synthetic residuals on site. In an attempt to answer this urgency, here a miniaturized fluorometric aptasensor prototype was contrived that catered to the point-of-care screening norm for two typical additives: chloramphenicol and enrofloxacin. The key target-indicating module worked in vitro based on the competitive binding-induced fluorescence recovery of fluorescein-labeled aptamers, which were photobleached beforehand in the format of double helix on burlike nanogold carriers. The “prickly” geometry of the latter not just enriched the capture probes at preferentially substrate-accessible spires; but also contributed to a tip-enhanced surface plasmon effect, sensitizing the signal-on during the duplex dissociation even at nanomolar threshold of the analytes. On the other hand, to encompass a full portable, a set of optical devices were mounted within a 3D-printed cartridge (adaptor) to converge the light beam and route it towards the detector, for which the smartphone camera came up in handy with a home-developed App for calibrating the emissive brightness. Enlightened by the high-dynamic-range compression, an imaging diagnostic algorithm was built in to grid and digitize each slide in the album for augmented detection performance. Thus, a novel bio-to-silico integration was invented that capable of in situ rapid reporting on the antibiotic presence with high sensitivity and selectivity. Further field practices in spiked milk on sales proved the precision and rudimentary feasibility of the well-assembled model of appliance, thus holding nice prospects in nonexpert (e.g., family and local community) utilities for foodborne antibiotic identification.

3D printed cartridge for high-speed capillary electrophoresis with sheath liquid thermostatting and contactless conductivity detection

The high-speed capillary electrophoresis (HSCE) method is a technique that utilizes a high electric field strength applied through a short capillary to reduce the time required for sample separation. However, the increased electric field strength may result in pronounced Joule heating effects. To address this, we describe a 3D-printed cartridge with integrated contactless conductivity detection (C4D) head and a sheath liquid channel. The C4D electrodes and Faraday shield layers are fabricated by casting Wood's metal in chambers inside the cartridge. Effective thermostatting of the short capillary is achieved by flowing Fluorinert liquid, which provides better heat dissipation compared to airflow. A HSCE device is created by using the cartridge and a modified slotted-vial array sample-introduction approach. Analytes are introduced through electrokinetic injection. With the help of sheath liquid thermostatting, background electrolyte concentration can be increased to several hundred mM, resulting in improved sample stacking and peak resolutions. Additionally, the baseline signal is flattened. Typical cations such as NH4+, K+, Na+, Mg2+, Li+, and Ca2+ can be separated within 22 s with an applied field strength of 1200 V/cm. The limit of detection ranges from 2.5 to 4.6 μM with a relative standard deviation of migration times of 1.1–1.2% (n = 17). The method has been applied to detect cations in drinking water and black tea leaching for drink safety testing, and to identify explosive anions in paper swabs. Samples can be directly injected without the need for dilution.

(Digital Presentation) Development of an Electrochemical Microfluidic Device with on-Platform Sample Collection

As the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to develop, the need for portable rapid testing platforms remains prevalent to provide patients with accurate and quantitative diagnostic and serosurveillance information at the point-of-care. The current gold standard detection techniques like RT-PCR and ELISA require trained personnel to perform lengthy protocols, resulting in a long turnover from sample collection to result acquisition. Herein, we propose an electrochemical microfluidic device for on-platform detection of viral proteins and antibodies at the point-of-care in a multiplexed manner. Miniaturization technology through the use of microfluidic devices offers numerous advantages including low reagent consumption, high fluidic control, reduced reaction times, inexpensive applications, and the possibility of throughput analysis. Electrochemical detection can provide advantages in cost effective fabrication, high sensitivity and simple instrumentation using a standard 3-electrode (working, reference, and counter) setup. Our platform proposes the design of an electrochemical cell with an enhanced working electrode to act as the detection assay with microfluidic channels to facilitate sample collection and pre-treatment; an integrated saliva collection kit and lancing device enabled the use of both untreated saliva from direct self-collection and whole blood from a finger prick. Automated fluid manipulation reduced the potential of user contamination through the implementation of suction-based flow. The electrochemical microfluidic device was encased in a 3D-printed cartridge for the fabrication of a fully integrative technology on a single platform with the potential to be used at the point-of-care in both clinical and commercial applications using direct biofluids.

Drug preconcentration and direct quantification in biofluids using 3D-Printed paper cartridge

Drug detection in biofluids has always been great importance for clinical diagnosis. Many detection technologies such as chromatography-mass spectrometry, have been applied to the detection of drugs. However, these technologies required multi-step operations, including complicated and time-consuming pretreatment processes and operations of bulky detection instruments, significantly limiting development of drug detection in clinical diagnosis. Herein, a portable 3D-printed paper cartridge was fabricated for fast sample preconcentration and direct drugs quantitative detection in biofluids by a portable Raman spectrometer. This cartridge contained both paper tip with silver nanowires to preconcentrate samples and achieve surface-enhanced Raman Scattering (SERS) measurement, and 3D-printed cartridge to build enclosed environment for the improvement of detection, which cost only one dollar. The preconcentration ability of the cartridge was up to 18.13-fold fluorescence enhancement and compared to the non-preconcentration method, it achieved 9.93-fold improvement of SERS performance. The anticancer drug of epirubicin hydrochloride, cyclophosphamide and their mixtures were quantitatively detected in the bovine serum or artificial urine. The integrated detection procedure required only 1 h, including sample pretreatment and preconcentration, drying, SERS measurements, and quantification analysis. This 3D-printed paper cartridge constituted a portable detection platform that would be potentially a practical and point-of-care detection tool for drug preconcentration and quantification on the clinical diagnosis.

Isolation of pathogenic bacteria from sputum samples using a 3D-printed cartridge system.

Within this contribution we introduce a 3D-printed cartridge system enabling the convenient and cost-efficient sample preparation from sputum for subsequent PCR based detection schemes. The developed fluidic system operates on pneumatic actuations. The closed system ensures a very low probability for contamination during sample processing, which is crucial when using a highly sensitive detection method such as PCR. The enrichment of the bacterial cells is achieved using different types of amine-functionalized particles. Our particle-based sample preparation approach yields intact and viable bacterial cells. Accordingly, not only PCR-based detection schemes can be employed, but also spectroscopic methods and biochemical tests, which require cultivation steps, are possible. The cartridge design in principle is compatible with magnetic and non-magnetic particle types. We investigated both variants and found that the performance of expanded glass beads is superior over the magnetic particles within the cartridge. Owing to the rather large size of the expanded glass beads, the dimensions of the channels can be enlarged, leading to lower hydrodynamic resistances, which is beneficial when processing viscous samples such as sputum. We verified the performance of our system using both artificial and real sputum samples containing Escherichia coli and Moraxella catarrhalis.

3D-printed cartridge system for in-flow photo-oxygenation of 7-aminothienopyridinones

A 3D-printed polypropylene (PP) continuous-photoflow cell based on a modular cartridge system was developed for the photo-oxygenation of 7-aminothieno[3,2-c]pyridin-4(5H)-ones, using ambient air as the sole co-reactant. This strategy takes advantage of the versatility of 3D-printing to construct cost-effective meso-scale reactors. In addition to scalability, a short residence time (tR 2 min) in 100-W blue LED light that minimizes the formation of dark, insoluble decomposition products is a tangible benefit of this technology.

Quadrupling the N95 Supply during the COVID-19 Crisis with an Innovative 3D-Printed Mask Adaptor.

The need for personal protective equipment during the COVID-19 pandemic is far outstripping our ability to manufacture and distribute these supplies to hospitals. In particular, the medical N95 mask shortage is resulting in healthcare providers reusing masks or utilizing masks with filtration properties that do not meet medical N95 standards. We developed a solution for immediate use: a mask adaptor, outfitted with a quarter section of an N95 respirator that maintains the N95 seal standard, thereby quadrupling the N95 supply. A variety of designs were 3D-printed and optimized based on the following criteria: seal efficacy, filter surface area and N95 respirator multiplicity. The final design is reusable and features a 3D-printed soft silicone base as well as a rigid 3D-printed cartridge to seal one-quarter of a 3M 1860 N95 mask. Our mask passed the computerized N95 fit test for six individuals. All files are publicly available with this publication. Our design can provide immediate support for healthcare professionals in dire need of medical N95 masks by extending the current supply by a factor of four.

Understanding and mitigating process drift in aerosol jet printing

Aerosol jet printing offers a versatile, high-resolution prototyping capability for flexible and hybrid electronics. Despite its rapid growth in recent years, persistent problems such as process drift hinder the adoption of this technology in production environments. Here we explore underlying causes of process drift during aerosol jet printing and introduce an engineered solution to improve deposition stability. It is shown that the ink level within the cartridge is a critical factor in determining atomization efficiency, such that the reduction in ink volume resulting from printing itself can induce significant and systematic process drift. By integrating a custom 3D-printed cartridge with an ink recirculation system, ink composition and level within the cartridge are better maintained. This strategy allows extended duration printing with improved stability, as evidenced by 30 h of printing over 5 production runs. This provides an important tool for extending the duration and improving reliability for aerosol jet printing, a key factor for integration in practical manufacturing operations.

Smartphone operable centrifugal system (SOCS) for on-site DNA extraction from foodborne bacterial pathogen.

The on-site recovery of nucleic acid from foodborne bacteria is in high demand to further understand on-site molecular diagnosis, which is especially applicable in developing countries. Here, we first proposed a smartphone operable centrifugal system (SOCS) for nucleic acid extraction with the assistance of a low power consumable motor and hydrogel beads. The SOCS consists of a centrifugal motor, 3D-printed cartridge, a nucleic acid collection column, and a smartphone. The SOCS shows excellent DNA extraction performance within 6 min, and it can operate more than 100 times using a smartphone. The purified effluent DNA was accumulated in the nucleic acid collection column. The performance of the SOCS was confirmed by amplifying the recovered DNA from Escherichia coli O157:H7. Moreover, the artificially inoculated food and blood samples also confirmed the performance of SOCS. The proposed SOCS provides an on-site operable nucleic acid separation platform in terms of simplicity, easy usability, cost-effectiveness, and portability in pathogenic point-of-care diagnostics.

Improving thermal control of capillary electrophoresis with mass spectrometry and capacitively coupled contactless conductivity detection by using 3D printed cartridges

A 3D-printed cartridge was developed to improve the interface between a capillary electrophoresis instrument and a mass spectrometer. The thermostated airflow from the CE was guided to the entrance of the electrospray source keeping as much as possible the silica capillary in a proper Joule-heating dissipation environment. Hollow 3D-printed walls made of ABS covered by a 0.2 mm thick copper foil on the inner side were used. The cartridge also allows including up to two capacitively coupled contactless conductivity detectors (C4Ds). Experiments about the separation of monoethyl carbonate (a thermally unstable species) shows that the peak area obtained with the original cartridge is only 21% of the value obtained with the 3D-printed cartridge, which demonstrates the improvement in heat dissipation.

3D-Printed Paper Spray Ionization Cartridge with Integrated Desolvation Feature and Ion Optics.

In this work we present the application of 3D-printing for the miniaturization and functionalization of an ion source for (portable) mass spectrometry (MS). Two versions of a 3D-printed cartridge for paper spray ionization (PSI) are demonstrated, assessed, and compared. We first focus on the use of 3D-printing to enable the integration of an embedded electrostatic lens and a manifold for internal sheath gas distribution and delivery. Cartridges with and without a sheath gas manifold and an electrostatic lens are compared with respect to analytical performance and operational flexibility. The sensitivity and limit of detection are improved in the cartridge with an electrostatic lens and sheath gas manifold compared to the cartridge without (15% and over 6.5× smaller, respectively). The use of these focusing elements also improved the average spray stability. Furthermore, the range of potentials required for PSI was lower, and the distance to the MS orifice over which spray could be obtained was larger. Importantly, both setups allowed quantification of a model drug in the ng/mL range with single-stage MS, after correction for spray instability. Finally, we believe that this work is an example of the impact that 3D-printing will have on the future of analytical device fabrication, miniaturization, and functionalization.

3D-printed paper spray ionization cartridge with fast wetting and continuous solvent supply features.

We report the development of a 3D-printed cartridge for paper spray ionization (PSI) that can be used almost immediately after solvent introduction in a dedicated reservoir and allows prolonged spray generation from a paper tip. The fast wetting feature described in this work is based on capillary action through paper and movement of fluid between paper and the cartridge material (polylactic acid, PLA). The influence of solvent composition, PLA conditioning of the cartridge with isopropanol, and solvent volume introduced into the reservoir have been investigated with relation to wetting time and the amount of solvent consumed for wetting. Spray has been demonstrated with this cartridge for tens of minutes, without any external pumping. It is shown that fast wetting and spray generation can easily be achieved using a number of solvent mixtures commonly used for PSI. The PSI cartridge was applied to the analysis of lidocaine from a paper tip using different solvent mixtures, and to the analysis of lidocaine from a serum sample. Finally, a demonstration of online paper chromatography-mass spectrometry is given.