APELLA: Open-Source, miniaturized All-in-One powered Lab-on-a-Disc platform.

We present an unconventional approach to a common Lab-on-a-Disc (LoD) that combines a quadcopter propulsion system, a miniaturized 2.4 GHz Wi-Fi spy camera, 9.74 Watt Qi wireless power, and an Arduino into an open-source, miniaturized All-in-one powered lab-on-disc platform (APELLA). The quadcopter propulsion generates thrust to rotate (from 0.1 to 24.5 Hz) or shake the LoD device, while the spy camera enables a real-time (30 frames per second) and high definition (1280 × 720 pixels) visualization of microfluidic channels without requiring a bulky and heavy stroboscopic imaging setup. A mobile device can communicate with an Arduino microcontroller inside the APELLA through a Bluetooth interface for closed loop and sequential frequency control. In a proof-of-concept study, the APELLA achieved comparable mixing efficiency to a traditional spin stand and can capture rapid microfluidic events at low rotational frequencies (<5Hz). The APELLA is low-cost (c.a. 100 Euro), compact (15.6 × 15.6 × 10 cm3), lightweight (0.59 kg), portable (powered by a 5 V USB power bank), and energy efficient (uses < 6% power of the conventional system), making it ideal for field deployment, education, resource-limited labs.

Moving perfusion culture and live-cell imaging from lab to disc: proof of concept toxicity assay with AI-based image analysis.

In vitro, cell-based assays are essential in diagnostics and drug development. There are ongoing efforts to establish new technologies that enable real-time detection of cell-drug interaction during culture under flow conditions. Our compact (10 × 10 × 8.5 cm) cell culture and microscope on disc (CMoD) platform aims to decrease the application barriers of existing lab-on-a-chip (LoC) approaches. For the first time in a centrifugal device, (i) cells were cultured for up to six days while a spindle motor facilitated culture medium perfusion, and (ii) an onboard microscope enabled live bright-field imaging of cells while the data wirelessly transmitted to a computer. The quantification of cells from the acquired images was done using artificial intelligence (AI) software. After optimization, the obtained cell viability data from the AI-based image analysis proved to correlate well with data collected from commonly used image analysis software. The CMoD was also suitable for conducting a proof-of-concept toxicity assay with HeLa cells under continuous flow. The half-maximal inhibitory time (IT50) for various concentrations of doxorubicin (DOX) in the case of HeLa cells in flow, was shown to be lower than the IT50 obtained from a static cytotoxicity assay, indicating a faster onset of cell death in flow. The CMoD proved to be easy to handle, enabled cell culture and monitoring without assistance, and is a promising tool for examining the dynamic processes of cells in real-time assays.

3D-Printed Radiopaque Microdevices with Enhanced Mucoadhesive Geometry for Oral Drug Delivery.

During the past decades, microdevices have been evaluated as a means to overcome challenges within oral drug delivery, thus improving bioavailability. Fabrication of microdevices is often limited to planar or simple 3D designs. Therefore, this work explores how microscale stereolithography 3D printing can be used to fabricate radiopaque microcontainers with enhanced mucoadhesive geometries, which can enhance bioavailability by increasing gastrointestinal retention. Ex vivo force measurements suggest increased mucoadhesion of microcontainers with adhering features, such as pillars and arrows, compared to a neutral design. In vivo studies, utilizing planar X-ray imaging, show the time-dependent gastrointestinal location of microcontainers, whereas computed tomography scanning and cryogenic scanning electron microscopy reveal information about their spatial dynamics and mucosal interactions, respectively. For the first time, the effect of 3D microdevice modifications on gastrointestinal retention is traced in vivo, and the applied methods provide a much-needed approach for investigating the impact of device design on gastrointestinal retention.

Real-Time Reflectance Measurement Using an Astigmatic Optical Profilometer.

An astigmatic optical profilometer with a commercial optical pickup head provides benefits, such as high resolution, compact size, and low cost. To eliminate artifacts caused by complex materials with different reflectances, a z-axis modulation mode is proposed to obtain quantitative surface morphology by measuring S curves on all image pixels. Moreover, the slope of the linear region in the S curve shows a positive relationship with the surface reflectance. However, the slope was calculated using an offline curve fitting method, which did not allow real-time reflectance imaging. Furthermore, quantitative reflectance data were unavailable because of the lack of calibration. In this study, we propose a novel method for real-time reflectance imaging by measuring the amplitude of a focus error signal (FES). The calibration results displayed a linear relationship between the FES amplitude and reflectance. The reflectance image of a grating sample with chrome patterns on a glass substrate demonstrates accurate reflectance measurements with a micrometer spatial resolution.

Low-cost, open-source XYZ nanopositioner for high-precision analytical applications.

Nanoscale positioning has numerous applications in both academia and industry. A growing number of applications require devices with long working distances and nanoscale resolutions. Friction-inertia piezoelectric positioners, which are based on the stick-slip mechanism, achieve both nanometer resolution and centimeter-scale travel. However, the requirements of complex preload mechanism, precision machining, and precise assembly increase the cost of conventional friction-inertia nanopositioners. Herein we present the design of an open-source XYZ-axis nanopositioning system. Utilizing a magnet-based stick-slip driving mechanism, the proposed XYZ nanopositioner provides several advantages, including sub-nanometer resolution, a payload capacity of up to 12 kg (horizontal), compact size, low cost, and easy assembly; furthermore, the system is adjustment-free. The performance tests validate the precision of the system in both scanning and stepping operation modes. Moreover, the resonant spectra affirm the rigidity and dynamic response of the mechanism. In addition, we demonstrate the practical applications of this nanopositioner in various measurement techniques, including scanning electron microscopy, vibrometry, and atomic force microscopy. Furthermore, we present 11 variations of the nanopositioner designs that are either compatible with ultra-high-vacuum systems and other existing systems, 3D printable, or hacking commercial linear slides.

In-Silico Selection of Aptamer Targeting SARS-CoV-2 Spike Protein.

Aptamers are single-stranded, short DNA or RNA oligonucleotides that can specifically bind to various target molecules. To diagnose the infected cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in time, numerous conventional methods are applied for viral detection via the amplification and quantification of DNA or antibodies specific to antigens on the virus. Herein, we generated a large number of mutated aptamer sequences, derived from a known sequence of receptor-binding domain (RBD)-1C aptamer, specific to the RBD of SARS-CoV-2 spike protein (S protein). Structural similarity, molecular docking, and molecular dynamics (MD) were utilized to screen aptamers and characterize the detailed interactions between the selected aptamers and the S protein. We identified two mutated aptamers, namely, RBD-1CM1 and RBD-1CM2, which presented better docking results against the S protein compared with the RBD-1C aptamer. Through the MD simulation, we further confirmed that the RBD-1CM1 aptamer can form the most stable complex with the S protein based on the number of hydrogen bonds formed between the two biomolecules. Based on the experimental data of quartz crystal microbalance (QCM), the RBD-1CM1 aptamer could produce larger signals in mass change and exhibit an improved binding affinity to the S protein. Therefore, the RBD-1CM1 aptamer, which was selected from 1431 mutants, was the best potential candidate for the detection of SARS-CoV-2. The RBD-1CM1 aptamer can be an alternative biological element for the development of SARS-CoV-2 diagnostic testing.

Open-source force analyzer with broad sensing range based on an optical pickup unit.

In the pharmaceutical field, oral drug delivery devices continue to shrink down to the micrometer scale, driving a trending demand to investigate ex vivo mucoadhesive force down to the micro-Newton scale. However, owing to the limitation of measuring sensitivity, conventional methods (e.g., a texture analyzer) lack reliability while measuring forces in this range. Herein, we report on an open-source force analyzer that utilizes an optical-pickup-unit (from a DVD player) to detect cantilever-based force transducers and thereby, achieves a wide force-sensing range from 1.1 N to 0.99 nN. The cantilever force transducers can easily be adjusted to fit different force ranges by adjusting the steel shim, magnets, and 3D printed components. To validate the analyzer, we conducted a preliminary study to investigate the effect of time and humidity of mucoadhesion of porcine intestinal tissues. Besides, we measured the mucoadhesive force of a single oral drug delivery microdevice with an average force of 93.7 µN on the top sides of the device. This analyzer offers the possibility of measuring e.g. mucoadhesion of individual microdevices in the micro-Newton range. Hence, the analyzer can assist in the development of miniaturized oral drug delivery devices but has a much wider field of potential force sensing applications.

Open source anaerobic and temperature-controlled in vitro model enabling real-time release studies with live bacteria.

In vitro release and dissolution models are widely used in the development phases of oral drug delivery systems to measure how an active pharmaceutical ingredient (API) is released from a dosage form. However, additional requirements for these models arise when evaluating probiotic dosage forms since they are often sensitive to temperature and oxygen levels. As a solution to this, we propose a custom-designed anaerobic in vitro release setup, made mainly by 3D printing and laser cutting, to function together with state-of-the-art pharmaceutical dissolution equipment - in this case, a microDISS Profiler™. The in vitro release model makes it possible to study the release rate of oxygen-sensitive probiotics in simulated intestinal conditions, while ensuring their survival due to the anaerobic conditions. This has not been possible so far since the available in vitro dissolution models have not been compatible with anaerobic conditions. With two different case studies, the developed model combined with a microDISS Profiler™ has proven capable of measuring the release of a probiotic and a small-molecule API from microdevices for oral drug delivery. Further, the model facilitated the survival of anaerobic bacteria present in the release medium.

Method for Film Thickness Mapping with an Astigmatic Optical Profilometer.

An astigmatic optical profilometer is a precision instrument with advantages such as high resolution, high bandwidth, a compact size, and low cost. However, current astigmatic optical profilometers measure only surface morphology, and their potential for capturing subsurface information remains underutilized. In this study, we developed a method for measuring the thickness of transparent thin films with an astigmatic optical profilometer. Experimental results demonstrate that the thickness of transparent films tens of micrometers thick can be accurately measured. The maximum thickness measurable through our system is approximately 100 µm, which may be increased to 1.2 mm through the use of a scanner with a greater travel range. A coupling problem occurs for films <25 µm in thickness. However, to solve this problem, we devised a decoupling method, which was experimentally implemented to successfully measure a 18-µm-thick film. Moreover, the ability to obtain 3D images, including of both the upper and lower surfaces, was demonstrated.

Consumer-Grade Inkjet Printer for Versatile and Precise Chemical Deposition.

Two simple, mechanical modifications are introduced to a consumer-grade inkjet printer to greatly increase its applicability. First, roller isolation bars are added to unlock multiple prints on the same substrate without smearing. This enables printing on a diverse set of substrates (rigid, elastic, liquid, granular, and sticky). Second, spring loadings are added to increase the print precision up to 50-fold, which facilitates alignment to a pre-patterned substrate or between successive prints. Utilizing the expanded substrate compatibility and the increased print precision, we explore tunable loading of drug combinations into microdevices. This loading method has promising applications within point-of-care personalized medication. Furthermore, we show how inkjet printers with array-type printheads (in our case, 6 x 90 nozzles) allow for quasi-simultaneous loading of reactants into microfluidic systems. The ability to do a quasi-simultaneous introduction of chemicals may be particularly useful for studies of rapidly reacting systems of three or more reactants, where premature introduction can shift the initial conditions from the intended. We believe that our modifications to an affordable system will inspire researchers to explore the possibilities of inkjet printing even further.

Quantifying Optical Absorption of Single Plasmonic Nanoparticles and Nanoparticle Dimers Using Microstring Resonators.

The wide and ever-increasing applications of thermoplasmonics demand the need for sensitive and reliable tools to probe optical absorptions of individual nanoparticles. However, most of the currently available techniques focus only on measuring the surface temperature of nanostructures in a particular medium and are either invasive or suffer from low sensitivity, lengthy calibration, or the inability to probe single structures with nanogaps. Here, we present for the first time the use of micromechanical SiN string resonators for quantifying optical absorption cross sections of individual plasmonic nanostructures. Monomers and dimers of nanospheres, nanostars, shell-isolated nanoparticles, and nanocubes are probed. A reliable data treatment method is developed to obtain the absorption cross sections as a function of responsivity across a string. The presented method exhibits an excellent sensitivity of ∼89 Hz/K. This allows quantification of optical absorption cross sections of individual plasmonic structures even when their plasmon resonance wavelengths are far from the laser excitation wavelength. The experimentally obtained optical absorption cross sections agree well with the simulations. Influencing factors including polarization, surface morphology, and nanogap size are discussed. The developed method and the obtained optical absorption profiles facilitate future development and optimization of thermoplasmonic applications.

Direct Measurement of Lateral Molecular Diffusivity on the Surface of Supersaturated Amorphous Solid Dispersions by Atomic Force Microscopy.

Quantifying molecular surface diffusivity is of broad interest in many different fields of science and technology. In this study, the method of surface grating decay is utilized to investigate the surface diffusion of practical relevant amorphous solid dispersions of indomethacin and the polymeric excipient Soluplus (a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer) at various polymer concentrations (1-20% w/w). The study confirms that measuring surface diffusivity below the system's glass transition temperature is possible with a simplified atomic force microscopy setup. Results highlight a striking polymer influence on the surface diffusivity of drug molecules at low polymer concentrations and a turnover point to a polymer dominated diffusion at around three percent (w/w) polymer concentration. The surface diffusion measurements further correlate well with the observed increase in physical stability of the system as measured by X-ray powder diffraction. These findings are of vital interest in both the applied use and fundamental understanding of amorphous solid dispersions.

Modular, Lightweight, Wireless Potentiostat-on-a-Disc for Electrochemical Detection in Centrifugal Microfluidics.

Interfacing electrochemical sensors in a lab-on-a-disc (LoD) system with a potentiostat is often tedious and challenging. We here present the first multichannel, modular, lightweight, and wirelessly powered, custom-built potentiostat-on-a-disc (PoD) for centrifugal microfluidic applications. The developed potentiostat is in the form factor of a typical digital video disc (DVD) and weighs only 127 g. The design of the potentiostat facilitates easy and robust interfacing with the electrodes in the LoD system, while enabling real-time electrochemical detection during rotation. The device can perform different electroanalytical techniques such as cyclic voltammetry, square wave voltammetry, and amperometry while being controlled by custom-made software. Measurements were conducted with and without rotation using both in-house fabricated and commercial electrodes. The performance of the PoD was in good agreement with the results obtained using a commercial potentiostat with a measured current resolution of 200 pA. As a proof of concept, we performed a real-time release study of an electrochemically active compound from microdevices used for drug delivery.

Ultrasensitive Microstring Resonators for Solid State Thermomechanical Analysis of Small and Large Molecules.

Thermal analysis plays an important role in both industrial and fundamental research and is widely used to study thermal characteristics of a variety of materials. However, despite considerable effort using different techniques, research struggles to resolve the physicochemical nature of many thermal transitions such as amorphous relaxations or structural changes in proteins. To overcome the limitations in sensitivity of conventional techniques and to gain new insight into the thermal and mechanical properties of small- and large-molecule samples, we have developed an instrumental analysis technique using resonating low-stress silicon nitride microstrings. With a simple sample deposition method and postprocess data analysis, we are able to perform rapid thermal analysis of direct instrumental triplicate samples with only pico- to nanograms of material. Utilizing this method, we present the first measurement of amorphous alpha and beta relaxation, as well as liquid crystalline transitions and decomposition of small-molecule samples deposited onto a microstring resonator. Furthermore, sensitive measurements of the glass transition of polymers and yet unresolved thermal responses of proteins below their apparent denaturation temperature, which seem to include the true solid state glass transition of pure protein, are reported. Where applicable, thermal events detected with the setup were in good agreement with conventional techniques such as differential scanning calorimetry and dynamic mechanical analysis. The sensitive detection of even subtle thermal transitions highlights further possibilities and applications of resonating microstrings in instrumental physicochemical analysis.

New Evidence for the Mechanism of Action of a Type-2 Diabetes Drug Using a Magnetic Bead-Based Automated Biosensing Platform.

The mechanism of action (MOA) of the first line type-2 diabetes drug metformin remains unclear despite its widespread usage. However, recent evidence suggests that the mitochondrial copper (Cu)-binding action of metformin may contribute toward the drug's MOA. Here, we present a novel biosensing platform for investigating the MOA of metformin using a magnetic microbead-based agglutination assay which has allowed us to demonstrate for the first time the interaction between Cu and metformin at clinically relevant low micromolar concentrations of the drug, thus suggesting a potential pathway of metformin's blood-glucose lowering action. In this assay, cysteine-functionalized magnetic beadswere agglutinated in the presence of Cu due to cysteine's Cu-chelation property. Addition of clinically relevant doses of metformin resulted in disaggregation of Cu-bridged bead-clusters, whereas the effect of adding a closely related but blood-glucose neutral drug propanediimidamide (PDI) showed completely different responses to the clusters. The entire assay was integrated in an automated microfluidics platform with an advanced optical imaging unit by which we investigated these aggregation-disaggregation phenomena in a reliable, automated, and user-friendly fashion with total assay time of 17 min requiring a sample (metformin/PDI) volume of 30 µL. The marked difference of Cu-binding action between the blood-glucose lowering drug metformin and its inactive analogue PDI thus suggests that metformin's distinctive Cu-binding properties may be required for its effect on glucose homeostasis. The novel automated platform demonstrating this novel investigation thus holds the potential to be utilized for investigating significant and sensitive molecular interactions via magnetic bead-based agglutination assay.

Optical imaging module for astigmatic detection system.

In this paper, an optical imaging module design for an astigmatic detection system (ADS) is presented. The module is based on a commercial optical pickup unit (OPU) and it contains a coaxial illuminant for illuminating a specimen. Furthermore, the imaging module facilitates viewing the specimen and the detection laser spot of the ADS with a lateral resolution of approximately 1 µm without requiring the removal of an element of the OPU. Two polarizers and one infrared filter are used to eliminate stray laser light in the OPU and stray light produced by the illuminant. Imaging modules designed for digital versatile disks (DVDs) and Blu-ray DVDs were demonstrated. Furthermore, the module can be used for imaging a small cantilever with approximate dimensions of 2 µm (width) × 5 µm (length), and therefore, it has the potential to be used in high-speed atomic force microscopy.

Blu-ray based optomagnetic aptasensor for detection of small molecules.

This paper describes an aptamer-based optomagnetic biosensor for detection of a small molecule based on target binding-induced inhibition of magnetic nanoparticle (MNP) clustering. For the detection of a target small molecule, two mutually exclusive binding reactions (aptamer-target binding and aptamer-DNA linker hybridization) are designed. An aptamer specific to the target and a DNA linker complementary to a part of the aptamer sequence are immobilized onto separate MNPs. Hybridization of the DNA linker and the aptamer induces formation of MNP clusters. The target-to-aptamer binding on MNPs prior to the addition of linker-functionalized MNPs significantly hinders the hybridization reaction, thus reducing the degree of MNP clustering. The clustering state, which is thus related to the target concentration, is then quantitatively determined by an optomagnetic readout technique that provides the hydrodynamic size distribution of MNPs and their clusters. A commercial Blu-ray optical pickup unit is used for optical signal acquisition, which enables the establishment of a low-cost and miniaturized biosensing platform. Experimental results show that the degree of MNP clustering correlates well with the concentration of a target small molecule, adenosine triphosphate (ATP) in this work, in the range between 10µM and 10mM. This successful proof-of-concept indicates that our optomagnetic aptasensor can be further developed as a low-cost biosensing platform for detection of small molecule biomarkers in an out-of-lab setting.

Quantification of rolling circle amplified DNA using magnetic nanobeads and a Blu-ray optical pick-up unit.

We present the first implementation of a Blu-ray optical pickup unit (OPU) for the high-performance low-cost readout of a homogeneous assay in a multichamber microfluidic disc with a chamber thickness of 600 µm. The assay relies on optical measurements of the dynamics of magnetic nanobeads in an oscillating magnetic field applied along the light propagation direction. The laser light provided by the OPU is transmitted through the sample chamber and reflected back onto the photo detector array of the OPU via a mirror. Spectra of the 2nd harmonic photo detector signal vs. the frequency of the applied magnetic field show a characteristic peak due to freely rotating magnetic nanobeads. Beads bound to ~1 µm coils of DNA formed off-chip by padlock probe recognition and rolling circle amplification show a different dynamics and the intensity of the characteristic peak decreases. We have determined the optimum magnetic bead concentration to 0.1mg/mL and have measured the response vs. concentration of DNA coils formed from Escherichia Coli. We have found a limit of detection of 10 pM and a dynamic range of about two orders of magnitude, which is comparable to the performance obtained using costly and bulky laboratory equipment. The presented device leverages on the advanced but low-cost technology of Blu-ray OPUs to provide a low-cost and high-performance magnetic bead-based readout of homogeneous bioassays. The device is highly flexible and we have demonstrated its use on microfluidic chambers in a disc with a thickness compatible with current optical media mass-production facilities.

Estimating Young's modulus of graphene with Raman scattering enhanced by micrometer tip.

We demonstrate that the Raman intensities of G and 2D bands of a suspended graphene can be enhanced using a gold tip with an apex size of 2.3 µm. The enhancement decays with the tip-graphene distance exponentially and remains detectable at a distance of 1.5 µm. Raman mappings show that the enhanced area is comparable to the apex size. Application of a bias voltage to the tip can attract the graphene so that Raman signals are intensified. The exponential enhancement-distance relationship enables the measurement of the graphene deformation, and the Young's modulus of graphene is estimated to be 1.48 TPa.

A compact synchrotron-based transmission X-ray microscope.

A compact transmission X-ray microscope has been designed and implemented based on a cylindrical symmetry around the optical axis that sharply limits the instabilities due to thermal mechanical drift. Identical compact multi-axis closed-loop actuation modules drive different optical components. The design is modular and simplifies the change of individual parts, e.g. the use of different magnification and focusing devices. This compact instrument can be easily transported between laboratory and synchrotron facilities and quickly put into operation. An automated alignment mechanism simplifies the assembly of different modules after transportation. After describing the design details, the results of the first tests are presented.