THz spectroscopy is an emerging technique for studying the dynamics and interactions of cells and biomolecules, but many practical challenges still remain in experimental studies. We present a prototype of simple and inexpensive cell-trapping microfluidic chip for THz spectroscopic study of live cells. Cells are transported, trapped and concentrated into the THz exposure region by applying an AC bias signal while the chip maintains a steady temperature at 37 degrees C by resistive heating. We conduct some preliminary experiments on E. coli and T-cell solution and compare the transmission spectra of empty channels, channels filled with aqueous media only, and channels filled with aqueous media with un-concentrated and concentrated cells.

Cells are the smallest living units and together they form all living organisms on earth. The cells are not only the building blocks of all living things, they also possess the most important information about life. A deeper understanding of these units may reveal hidden secrets about difficultly cured diseases, memory and learning, among others. Today’s techniques have problems such as low sensitivity, lethal preparation steps for the cells and overlaps in result spectra. Microfluidics has shown to be a useful tool allowing improved dynamic control, high throughput and sensitivity in nanoliters. The aim with this project is to design a microfluidic system for future integration with photonic sensors. Three different designs were developed, one design with the aim to integrate with photonic sensors and two for cell trapping only. Simulations and analytical calculations were performed to verify the requirements of single cell trapping. Simulation and analytical calculation results consorted, except for the ladder design. Moreover, strength calculations were performed for the sensor, to verify that it could handle the high pressures. A fabrication process was developed and an OSTE polymer was chosen as a suitable material. The transparency of the OSTE for fluorescent signals was studied. Results from the fabrication show proper lithography and molding as well as flow through channels. However, bubbles tend to appear in the channels. A rough surface of the chip appeared to primarily come from defects and filth on mask and mold. Three different connector solutions were tested, but they could not stand the high pressures. The work in this project has taken the development one step closer to the final goal to integrate photonic biosensors with a microfluidic system enabling single cell sensing.

No description available.

This work presents detailed numerical calculations of the dielectrophoretic force in octupolar traps designed for single-cell trapping. A trap with eight planar electrodes is studied for spherical and ellipsoidal particles using an indirect implementation of the boundary element method (BEM). Multipolar approximations of orders one to three are compared with the full Maxwell stress tensor (MST) calculation of the electrical force on spherical particles. Ellipsoidal particles are also studied, but in their case only the dipolar approximation is available for comparison with the MST solution. The results show that the full MST calculation is only required in the study of non-spherical particles. / Singapore-MIT Alliance (SMA)

No description available.

Les agrégats cellulaires constituent un modèle intermédiaire entre les cellules uniques et les tissues cellulaires et sont utilisés dans de nombreux domaines tels que l’ingénierie tissulaire et le criblage de médicaments in vitro. La création de tels agrégats cellulaires dont les propriétés et la taille seraient contrôlées nécessite cependant le développement de nouvelles approches ascendantes. Le travail présenté dans ce manuscrit vise à développer des microsystèmes pour la formation contrôlée d’agrégats de cellules sous flux via des champs électriques. Cette approche se base sur la diélectrophorèse (DEP), un phénomène induisant le déplacement des particules diélectriques lorsqu’elles sont placées dans un champ électrique non-uniforme. Un outil de calcul, MyDEP, a tout d’abord été développé afin d’être en mesure de prédire le comportement des cellules en suspension dans un certain milieu. Cet outil permet d’étudier la réponse diélectrique des particules et des cellules en fonction de la fréquence du champ. Il contient une base de données regroupant les propriétés diélectriques des cellules publiées dans la littérature afin d’aider tant les spécialistes que les utilisateurs néophytes à comprendre le comportement diélectrophorétique des particules et des cellules ainsi qu’à choisir les paramètres expérimentaux tels que la conductivité électrique du milieu et la fréquence du champ préalablement aux manipulations expérimentales en laboratoire. Différents designs pour le piégeage de cellules sont proposés avec les simulations, par la méthode des éléments finis en utilisant COMSOL Multiphysics, associées. Leur fabrication a nécessité le développement d’une méthode d’alignement reproductible, précise au micromètre, des microcanaux d’un polymère appelé le polydiméthylsiloxane (PDMS) avec des électrodes coplanaires en titane/platine déposées sur du verre via l’utilisation d’une aligneuse de masques conventionnelle. La méthode est basée sur l’utilisation d’un moule en silicium associé à un sarcophage en Poly(methyl methacrylate) (PMMA) afin de garantir le contrôle du parallélisme entre les parties supérieure et inférieure du PDMS moulé. Les puces contenant les différents designs de piégeage ainsi fabriquées ont été testées avec succès sur des cellules rénales embryonnaires humaines (HEK) à l’aide d’une installation expérimentale démontrant par la même la capacité des puces à créer des agrégats constitués d’un nombre contrôlé de cellules par diélectrophorèse. Les agrégats ainsi formés se sont avérés stables après 5 minutes de contact cellule à cellule sans qu’une séparation des cellules n’ait été observée. Le design d’un capteur par impédance a par ailleurs été proposé pour caractériser tant les cellules uniques que les agrégats cellulaires avant et après la chambre de piégeage. Celui-ci, associé au design de piégeage par DEP, a été testé expérimentalement avec succès pour détecter leur passage. / Cell aggregates are an intermediary model between single cells and cell tissues used in many applications such as tissue engineering and in vitro drug screening. The creation of cells aggregates of controlled size and properties requires the development of new bottom-up strategies. The work developed in this manuscript aims at presenting the development of microsystems for the electric force-driven controlled formation of cell aggregates under flow conditions. This approach is based on dielectrophoresis, a phenomenon that causes induced motion on dielectric particles placed in a non-uniform electric field. A computational tool, MyDEP, was first developed in order to predict the behavior of cells in a specific medium. It allows to study the dielectric response of particles and cells as a function of frequency. The software also includes a database gathering cell dielectric models available in the literature to help experienced users as well as neophytes to understand the dielectrophoretic behavior of particles and cells and to choose parameters such as electric conductivity of the medium and frequency before performing laboratory experiments. Different designs for cell trapping are proposed and simulated in 2D with FEM using COMSOL Multiphysics. Their fabrication implied the development of a reproducible method for μm precision alignment of microchannels in a polymer called polydimethylsiloxane (PDMS) with coplanar titanium/platinum electrodes deposited on glass, using a conventional mask aligner. It is based on the use of a silicon mold in combination with a Poly(methyl methacrylate) (PMMA) sarcophagus for precise control of the parallelism between top and bottom surfaces of molded PDMS. The trapping design based on coplanar electrodes was successfully tested experimentally on human embryonic kidney cells (HEK) with an automated setup. It proves its capability to create aggregates of a controlled number of cells with DEP. The cell aggregates proved to be stable (no disruption) after only 5 minutes of cell-cell contact. An impedance-based sensor design was proposed for characterizing single cells and cells aggregates before and after the trapping chamber. This sensor was successfully tested experimentally to detect particle passage in combination with the dielectrophoretic trapping design.

In a time when microorganisms are controlling the world, research in biology is more relevant than ever and this requires some powerful instruments. Optical tweezers use a focused laser beam to manipulate and probe objects on the nano- and microscale. This allows for the exploration of a miniature world at the border between biology, chemistry and physics. New methods for biophysical and physicochemical measurements are continuously being developed and at Umeå University there is a need for a new system that combines several of these methods. This would truly be the new Swiss army knife of biophysical and biomechanical research, extending their reach in the world of optical tweezing. My ambition with this project is to design and construct a robust system that incorporates optical trapping with high-precision force measurements and Raman spectroscopy, as well as introducing the possibility of generating multiple traps by using a spatial light modulator (SLM). The proposed design incorporates four different lasers and a novel combination of signal detection techniques. To allow for precise control of the systems components and laser beams, I designed and constructed motorized opto-mechanical components. These are controlled by an in-house developed software that handles data processing and signal analysis, while also providing a user interface for the system. The components include, motorized beam blockers and optical attenuators, which were developed using commonly available 3D printing techniques and electronic controllers. By designing the system from scratch, I could eliminate the known weaknesses of conventional systems and allow for a modular design where components can be added easily. The system is divided into two parts, a laser breadboard and a main breadboard. The former contains all the equipment needed to generate and control the laser beams, which are then coupled through optical fibers to the latter. This contains the components needed to move the optical trap inside the sample chamber, while performing measurements and providing user feedback. Construction and testing was done for one sub-system at a time, while the lack of time required a postponement for the implementation of Raman and SLM. The system performance was verified through Allan variance stability tests and the results were compared with other optical tweezers setups. The results show that the system follows the thermal limit for averaging times (τ) up to ~1 s when disturbances had been eliminated, which is similar to other systems. However, we could also show a decrease in variance all the way to τ = 2000 s, which is exceptionally good and not found in conventional systems. The force-resolution was determined to be on the order of femtonewtons, which is also exceptionally good. Thus, I conclude that this optical tweezers setup could lie as a solid foundation for future development and research in biological science at Umeå University for years to come.