We demonstrate a microplate platform for parallelized manipulation of particles or cells by frequency-modulated ultrasound. The device, consisting silicon-glass microchip and single ultrasonic transducer, enables aggregation, positioning high-resolution microscopy distributed in an array 100 microwells centered on the microchip. characterize system terms temperature control, aggregation efficiency, cell viability. use time-lapse imaging to show that continuously exposed ultrasound are able...

We present an easy-to-use, disposable, thermoplastic microwell chip designed to support screening and high-resolution imaging of single-cell behavior in two- three-dimensional (2D 3D) cell cultures. show that the has excellent optical properties provide simple protocols for efficient long-term culture suspension adherent cells, latter grown either as monolayers or hundreds single, uniformly sized spheroids. then demonstrate applicability system analysis by correlating dynamic cytotoxic...

The primary cilium is a ubiquitous organelle present on most mammalian cells. Malfunction of the has been associated with various pathological disorders, many which lead to cystic disorders in liver, pancreas, and kidney. Primary cilia have kidney epithelial cells observed generate intracellular calcium response fluid flow, disruption proteins involved this signaling autosomal dominant polycystic disease, implying direct connection between cyst formation. It also shown that there significant...

Understanding the complex 3D tumor microenvironment is important in cancer research. This can be modelled vitro by culturing multicellular spheroids (MCTS). Key challenges when using MCTS applications such as high-throughput drug screening are overcoming imaging and analytical issues encountered during functional structural investigations. To address these challenges, we use an ultrasonic standing wave (USW) based culture platform for parallel formation, staining of 100 whole MCTS. A protein...

New markers are constantly emerging that identify smaller and subpopulations of immune cells. However, there is a growing awareness even within very small populations, marked functional heterogeneity measurements at the population level only gives an average estimate behaviour pool techniques to analyze single cells over time needed overcome this limitation. For purpose, we have designed evaluated microwell array systems made from two materials, polydimethylsiloxane (PDMS) silicon, for...

Journal Article Live cell imaging in a micro-array of acoustic traps facilitates quantification natural killer heterogeneity Get access Athanasia E. Christakou, Christakou Department Applied Physics, Albanova University Center, KTH - Royal Institute Technology, SE-10691, Stockholm, Sweden. Tel: +46 8 55378793, 730 766146 Search for other works by this author on: Oxford Academic Google Scholar Mathias Ohlin, Ohlin Bruno Vanherberghen, Vanherberghen Mohammad Ali Khorshidi, Khorshidi Nadir...

Here, we report a screening platform for assessment of the cytotoxic potential individual natural killer (NK) cells within larger populations. Human primary NK were distributed across silicon-glass microchip containing 32,400 microwells loaded with target cells. Through fluorescence and automated image analysis, numbers live or dead in each well could be assessed at different time points after initial mixing. Cytotoxicity was also studied by time-lapse live-cell imaging quantifying killing...

Immune cells exhibit functional heterogeneity beyond what is resolved by classical definitions of subpopulations based on cell surface expression receptors. To develop efficient and personalized cell-based immunotherapies, we need to resolve this understand the underlying parameters that dictate cellular responses specific target cells. For this, new methods are required can identify harvest immune with functions, e.g., high cytotoxic potential, form clonally expanded or assess molecular...

Each individual has a heterogeneous pool of NK cells consisting that may be specialized towards specific functional responses such as secretion cytokines or killing tumor cells. Many conventional methods are not fit to characterize populations they measure the average response all Thus, there is need for experimental platforms provide single cell resolution. In addition, also transient and stochastic variations in at level, calling allow studies many events over extended times. This paper...

We demonstrate a concept for how miniaturized 3-D cell culture in biological extracellular matrix (ECM) or synthetic gels bridges the gap between organ-tissue and traditional 2-D cultures. A microfluidic device including microgradient environments has been designed, fabricated, successfully evaluated. In presented system stable diffusion gradients can be generated by application of two parallel fluid flows with different composition against opposite sides gel plug embedded cells. Culture up...

This review describes the use of ultrasound for inducing and retaining cell-cell contact in multi-well microplates combined with live-cell fluorescence microscopy. platform has been used studying interaction between natural killer (NK) cells cancer at level individual cells. The includes basic principles ultrasonic particle manipulation, design criteria when building a microplate device this purpose, biocompatibility aspects, finally, two examples biological applications: Dynamic imaging...

We characterize and quantify the performance of ultrasonic particle aggregation positioning in a 100-well microplate. analyze result when operating planar ring transducer at different single actuation frequencies range 2.20–2.40 MHz, compare with obtained from schemes frequency-modulated actuation. Compared to our previously used wedge design, has larger contact area facing microplate, resulting lower temperature increase for given voltage. Furthermore, we dynamics acoustic streaming...

Glucose is a routine emergency sample. General guidelines for inpatient hyperglycemia are scarce, except in myocardial infarction, stroke, and perioperative/ICU. Previous studies found admission glucose associated with increased mortality specific conditions. Scandinavian data, general patients, scarcer. We investigated levels, 30-day mortality, length-of-stay (LoS), Swedish hospital.From 8146 visits data regarding age, gender, dates of admission, discharge death, diagnoses, p-glucose,...

Electron-beam lithography (EBL) is a relevant technique to the nanoscience community as it enables production of precise structures at nanoscale. When writing features in thick resist layer, dose insufficiency typically encountered when resolution approaches focal spot electron beam itself. We present study this phenomenon, theory for its understanding and compensation, method assignment correct area small features. Dose originates from proximity effect distributing energy volumes that are...

Here, we present a methodology based on multiplexed fluorescence screening of two- or three-dimensional cell cultures in newly designed multichambered microwell chip, allowing direct assessment drug immune cytotoxic efficacy. We establish framework for culture, formation tumor spheroids, labeling, and imaging fixed live cells at various magnifications directly the chip together with data analysis interpretation. The is demonstrated by cytotoxicity using ovarian non-small lung cancer cellular...

This paper presents a novel method to embed, anchor, and cultivate cells in controlled 3-D flow-through microenvironment. is realized using an etched silicon pillar flow chamber filled with extracellular matrix (ECM) gel mixed cells. At 4°C, while liquid form, ECM injected into the chamber. Raising temperature 37°C results gel, embedded. The pillars both stabilize increase surface volume ratio of gel. During polymerization shrinks, thus creating channels, which enables perfusion through...

A novel micromachined interface for airborne sample-to-liquid adsorption and droplet-to-liquid transfer was designed fabricated. It enables a robust sheet liquid flow serving as an site. The characterised pressure properties tested successfully the transfer/adsorption of different samples. qualitative theoretical model device characteristics is presented. We also used to introduce method system fast detection dust- vapour-based narcotics explosives traces. microfluidic vapour-to-liquid...

X-ray zone plates made from gold are common optical components used in imaging experiments. These nanostructures normally fabricated using a combination of electron-beam lithography and electroplating with cyanide baths. In this study, we present process miniaturized gold-suplphite bath. The miniaturization is enabled by on-chip reference plating areas well defined sizes, offering reliable way to control the height structures carefully choosing time at given current density accordance...

The spatial distribution of activating ligands dictates NK cell synapse formation, stability, and function.

Metal assisted chemical etching is a promising method for fabricating high aspect ratio micro- and nanostructures in silicon. Previous results have suggested that P-type N-type silicon etches with different degrees of anisotropy, questioning the use nanostructures. In this study, we compare processing X-ray zone plate N through metal gold catalyst. Fabricated plates were cleaved studied focus on resulting verticality, depth porosity. Results show nanostructures, both prove to be viable...

We present the design, fabrication and successful testing of a 14×14×4 mm3 integrated electronic narcotics sensing system which consists only four parts. The microsystem absorbs airborne molecules performs liquid assay using an quartz crystal microbalance (QCM). A vertically conductive double-sided adhesive foil (VCAF) was used studied as novel material for LOC MEMS applications provides easy assembly, electrical contacting containment. tested measuring cocaine ecstasy, with detection...

Abstract In this work, a new technique for off‐line hyphenation between CE and MALDI‐MS is presented. Two closed fused‐silica capillaries were connected via silicon chip comprising an open microcanal. The EOF in the system was evaluated using mesityloxide or leucine‐enkephalin as sample with running buffer that rendered analyte neutrally charged. Comparison made (first capillary solely included electrical circuit) closed–open microcanal circuit). It concluded experimental values of agreed...

Natural killer (NK) cell cytotoxicity in tissue is dependent on the ability of NK cells to migrate through extracellular matrix (ECM) microenvironment. Traditional imaging studies migration and have utilized 2D surfaces, which do not properly reproduce structural mechanical cues that shape migratory response vivo. Here, we combined a microwell assay allows long-term tracking small, well-defined populations with an interstitial ECM-like matrix. The for NK-target interactions within confined...

Zone plates are diffractive optics commonly used in X-ray microscopes. Here, we present a wet-chemical approach for fabricating high aspect ratio Pd/Si zone plate aimed at the hard regime. A Si mold is fabricated via metal-assisted chemical etching (MACE) and further metalized with Pd electroless deposition (ELD). MACE results vertical zones ratios. The observed rate our design 700 nm/min. ELD metallization yields density of 10.7 g/cm 3 , value slightly lower than theoretical 12 . Fabricated...