Surface acoustic wave microfluidic pumps for on-chip diagnostics

Most of point-of-care diagnostics and lab-on-chip devices that do on-chip sample preparation require active fluid actuation. In a laboratory setting, this is done via bulky benchtop equipment such as syringe pumps, peristaltic pumps and pressure systems. However, integration of a pumping unit onto the device allows for increased portability and decreased footprint of the device. Although there are multiple examples of realised micropumps based on different technologies, no one solution offers a combination of small footprint, low costs, scalable manufacturing and high performance required for point-of-care devices. Surface acoustic wave (SAW)-based micropumps are an exciting alternative to the current micropump systems due their small footprint and simplicity of manufacturing, yet many of the SAW micropumps presented to date suffer from poor performance and/or utilisation of open channels, which can be a problem regarding contamination. The SAW micropump demonstrated here uses a novel planar design and SAW scattering effects to significantly improve the pump performance and maintain closed channels, which is a pre-requisite for point-of-care applications. This thesis evaluates the fabrication of SAW devices and microfluidic channels using soft lithography. After evaluating the SAW device design concerning electrical characteristics both experimentally and theoretically, the first iteration of SAW micropumps utilising SAW momentum along the piezoelectric substrate is presented and characterised in terms of fluid flow velocity profiles and volume flow rates produced. Subsequently, a concept of a more efficient SAW micropump is presented based on out of the plane interaction between SAW and liquid. To fully utilise this interaction a protocol on the development of 3D microfluidic channels is introduced followed by a discussion on SAW-liquid coupling setting the scene for a demonstration of efficient and closed-loop SAW micropump that delivers pressure gradients up to an order of magnitude higher than the best to-date reported values at a similar input power levels. Finally, the newly developed pump is utilised in an on-chip flow cytometer to showcase the advanced flow manipulations, showing the potential applications of the SAW micropump beyond simple fluid actuation

Lung on a Chip Development from Off-Stoichiometry Thiol–Ene Polymer

Current in vitro models have significant limitations for new respiratory disease research and rapid drug repurposing. Lung on a chip (LOAC) technology offers a potential solution to these problems. However, these devices typically are fabricated from polydimethylsiloxane (PDMS), which has small hydrophobic molecule absorption, which hinders the application of this technology in drug repurposing for respiratory diseases. Off-stoichiometry thiol–ene (OSTE) is a promising alternative material class to PDMS. Therefore, this study aimed to test OSTE as an alternative material for LOAC prototype development and compare it to PDMS. We tested OSTE material for light transmission, small molecule absorption, inhibition of enzymatic reactions, membrane particle, and fluorescent dye absorption. Next, we microfabricated LOAC devices from PDMS and OSTE, functionalized with human umbilical vein endothelial cell (HUVEC) and A549 cell lines, and analyzed them with immunofluorescence. We demonstrated that compared to PDMS, OSTE has similar absorption of membrane particles and effect on enzymatic reactions, significantly lower small molecule absorption, and lower light transmission. Consequently, the immunofluorescence of OSTE LOAC was significantly impaired by OSTE optical properties. In conclusion, OSTE is a promising material for LOAC, but optical issues should be addressed in future LOAC prototypes to benefit from the material properties

NV microscopy of thermally controlled stresses caused by thin Cr2O3 films

Many modern applications, including quantum computing and quantum sensing, use substrate-film interfaces. Particularly, thin films of chromium or titanium and their oxides are commonly used to bind various structures, such as resonators, masks, or microwave antennas, to a diamond surface. Due to different thermal expansions of involved materials, such films and structures could produce significant stresses, which need to be measured or predicted. In this paper, we demonstrate imaging of stresses in the top layer of diamond with deposited structures of Cr2O3 at temperatures 19°C and 37°C by using stress-sensitive optically detected magnetic resonances (ODMR) in NV centers. We also calculated stresses in the diamond-film interface by using finite-element analysis and correlated them to measured ODMR frequency shifts. As predicted by the simulation, the measured high-contrast frequency-shift patterns are only due to thermal stresses, whose spin-stress coupling constant along the NV axis is 21±1 MHz/GPa, that is in agreement with constants previously obtained from single NV centers in diamond cantilever. We demonstrate that NV microscopy is a convenient platform for optically detecting and quantifying spatial distributions of stresses in diamond-based photonic devices with micrometer precision and propose thin films as a means for local application of temperature-controlled stresses. Our results also show that thin-film structures produce significant stresses in diamond substrates, which should be accounted for in NV-based applications. © 2023 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement. --//-- This is an open access article Andris Berzins, Janis Smits, Andrejs Petruhins, Roberts Rimsa, Gatis Mozolevskis, Martins Zubkins, and Ilja Fescenko, "NV microscopy of thermally controlled stresses caused by thin Cr2O3 films," Opt. Express 31, 17950-17963 (2023), https://doi.org/10.1364/OE.489901.Centrālā finanšu un līgumu aģentūra (CFLA) (2.3.1.1.i.0/1/22/I/CFLA/001); European Regional Development Fund (ERAF) (1.1.1.5/20/A/001); Horizon 2020 Framework Programme H2020-WIDESPREAD-01-2016-2017-Teaming Phase2 (739508, CAMART2); Latvijas Universitātes fonds ("Annealing furnace for the development of sensors", "Improvement of Magnetic field imaging system", "Simulations for stimulation of science"); Latvijas Zinātnes Padome (lzp-2020/2-0243, lzp-2021/1-0379); State Education Development Agency Republic of Latvia (1.1.1.2/VIAA/1/16/024)

Lung on a Chip Development from Off-Stoichiometry Thiol–Ene Polymer

Institute of Solid-State Physics, University of Latvia as the Center of Excellence has received funding from the European Union’s Horizon 2020 Framework Programme H2020-WIDESPREAD-01-2016-2017-TeamingPhase2 under grant agreement No. 739508, project CAMART2. Finally, we would like to thank Biol. Kaspars Tars from Latvian Biomedical research and study center for giving us the opportunity to participate in this consortium and contribute to Latvian scientists’ effort in response to the COVID-19 pandemic.Current in vitro models have significant limitations for new respiratory disease research and rapid drug repurposing. Lung on a chip (LOAC) technology offers a potential solution to these problems. However, these devices typically are fabricated from polydimethylsiloxane (PDMS), which has small hydrophobic molecule absorption, which hinders the application of this technology in drug repurposing for respiratory diseases. Off-stoichiometry thiol–ene (OSTE) is a promising alternative material class to PDMS. Therefore, this study aimed to test OSTE as an alternative material for LOAC prototype development and compare it to PDMS. We tested OSTE material for light transmission, small molecule absorption, inhibition of enzymatic reactions, membrane particle, and fluorescent dye absorption. Next, we microfabricated LOAC devices from PDMS and OSTE, functionalized with human umbilical vein endothelial cell (HUVEC) and A549 cell lines, and analyzed them with immunofluorescence. We demonstrated that compared to PDMS, OSTE has similar absorption of membrane particles and effect on enzymatic reactions, significantly lower small molecule absorption, and lower light transmission. Consequently, the immunofluorescence of OSTE LOAC was significantly impaired by OSTE optical properties. In conclusion, OSTE is a promising material for LOAC, but optical issues should be addressed in future LOAC prototypes to benefit from the material properties.--//--This work is licensed under a CC BY 4.0 license.This research was funded by project Nr. VPP-COVID-2020/1-0014 awarded by Latvian Council of Scienc

Extracellular Vesicles Isolation from Large Volume Samples Using a Polydimethylsiloxane-Free Microfluidic Device

Extracellular vesicles (EV) have many attributes important for biomedicine; however, current EV isolation methods require long multi-step protocols that generally involve bulky equipment that cannot be easily translated to clinics. Our aim was to design a new cyclic olefin copolymer–off-stoichiometry thiol-ene (COC–OSTE) asymmetric flow field fractionation microfluidic device that could isolate EV from high-volume samples in a simple and efficient manner. We tested the device with large volumes of urine and conditioned cell media samples, and compared it with the two most commonly used EV isolation methods. Our device was able to separate particles by size and buoyancy, and the attained size distribution was significantly smaller than other methods. This would allow for targeting EV size fractions of interest in the future. However, the results were sample dependent, with some samples showing significant improvement over the current EV separation methods. We present a novel design for a COC–OSTE microfluidic device, based on bifurcating asymmetric flow field-flow fractionation (A4F) technology, which is able to isolate EV from large volume samples in a simple, continuous-flow manner. Its potential to be mass-manufactured increases the chances of implementing EV isolation in a clinical or industry-friendly setting, which requires high repeatability and throughput

Bifurcated Asymmetric Field Flow Fractionation of Nanoparticles in PDMS-Free Microfluidic Devices for Applications in Label-Free Extracellular Vesicle Separation

Extracellular vesicles are small membrane-bound structures that are released by cells and play important roles in intercellular communication garnering significant attention in scientific society recently due to their potential as diagnostic and therapeutic tools. However, separating EVs from large-volume samples remains a challenge due to their small size and low concentration. In this manuscript, we presented a novel method for separating polystyrene beads as control and extracellular vesicles from large sample volumes using bifurcated asymmetric field flow fractionation in PDMS-free microfluidic devices. Separation characteristics were evaluated using the control system of polystyrene bead mix, which offers up to 3.7X enrichment of EV-sized beads. Furthermore, in the EV-sample from bioreactor culture media, we observed a notable population distribution shift of extracellular vesicles. Herein presented novel PDMS-free microfluidic device fabrication protocol resulted in devices with reduced EV-loss compared to size-exclusion columns. This method represented an improvement over the current state of the art in terms of EV separation from large sample volumes through the use of novel field flow fractionation design

Bifurcated Asymmetric Field Flow Fractionation of Nanoparticles in PDMS-Free Microfluidic Devices for Applications in Label-Free Extracellular Vesicle Separation

Extracellular vesicles are small membrane-bound structures that are released by cells and play important roles in intercellular communication garnering significant attention in scientific society recently due to their potential as diagnostic and therapeutic tools. However, separating EVs from large-volume samples remains a challenge due to their small size and low concentration. In this manuscript, we presented a novel method for separating polystyrene beads as control and extracellular vesicles from large sample volumes using bifurcated asymmetric field flow fractionation in PDMS-free microfluidic devices. Separation characteristics were evaluated using the control system of polystyrene bead mix, which offers up to 3.7X enrichment of EV-sized beads. Furthermore, in the EV-sample from bioreactor culture media, we observed a notable population distribution shift of extracellular vesicles. Herein presented novel PDMS-free microfluidic device fabrication protocol resulted in devices with reduced EV-loss compared to size-exclusion columns. This method represented an improvement over the current state of the art in terms of EV separation from large sample volumes through the use of novel field flow fractionation design

Rapid cell separation with minimal manipulation for autologous cell therapies

The ability to isolate specific, viable cell populations from mixed ensembles with minimal manipulation and within intra-operative time would provide significant advantages for autologous, cell-based therapies in regenerative medicine. Current cell-enrichment technologies are either slow, lack specificity and/or require labelling. Thus a rapid, label-free separation technology that does not affect cell functionality, viability or phenotype is highly desirable. Here, we demonstrate separation of viable from non-viable human stromal cells using remote dielectrophoresis, in which an electric field is coupled into a microfluidic channel using shear-horizontal surface acoustic waves, producing an array of virtual electrodes within the channel. This allows high-throughput dielectrophoretic cell separation in high conductivity, physiological-like fluids, overcoming the limitations of conventional dielectrophoresis. We demonstrate viable/non-viable separation efficacy of >98% in pre-purified mesenchymal stromal cells, extracted from human dental pulp, with no adverse effects on cell viability, or on their subsequent osteogenic capabilities.Published versio