Design of turbulent tangential micro-mixers that mix liquids on the nanosecond time scale

Unravelling (bio)chemical reaction mechanisms and macromolecular folding pathways on the (sub)microsecond time scale is limited by the time resolution of kinetic instruments for mixing reactants and observation of the progress of the reaction. To improve the mixing time resolution, turbulent four- and two-jet tangential micro-mixers were designed and characterized for their mixing and (unwanted) premixing performances employing acid-base reactions monitored by a pH-sensitive fluorescent dye. The mixing performances of the micro-mixers were determined after the mixing chamber in a free-flowing jet. The premixing behavior in the vortex chamber was assessed in an optically transparent glass-silicon replica of a previously well-characterized stainless-steel four-jet tangential micro-mixer. At the highest flow rates, complete mixing was achieved in 160 ns with only approximately 9% premixing of the reactants. The mixing time of 160 ns is at least 50 times shorter than estimated for other fast mixing devices. Key aspects to the design of ultrafast turbulent micro-mixers are discussed. The integration of these micro-mixers with an optical flow cell would enable the study of the very onset of chemical reactions in general and of enzyme catalytic reactions in particula

Field-effect based attomole titrations in nanoconfinement

This paper describes a novel capacitive method to change the pH in micro- and nanofluidic channels. A device with two metal gate electrodes outside an insulating channel wall is used for this purpose. The device is operated at high ionic strength with thin double layers. We demonstrate that gate potentials applied between the electrodes cause a release or uptake of protons from the silicon nitride surface groups, resulting in a pH shift in the channel and a titration of solution compounds present. Due to the high quality silicon nitride insulating layer, the effect is purely capacitive and electrolysis can be neglected. Fluorescein was employed as a fluorescent pH indicator to quantify the induced pH changes, and a maximum change of 1.6 pH units was calculated. A linear relationship was found between applied potential and fluorescein intensity change, indicating a linear relation between actuated proton amount and applied voltage. Since this pH actuation method avoids redox reactions and can be operated at physiological ionic strength, it can be very useful as a soft way to change the pH in very small volumes e.g. in bioassays or cell-based research. The sensitivity of the optical detection method poses the only limit to the detectable amount of substance and the observed volume. In a preliminary measurement we show one possible application, namely titration of 100 attomol of TRIS in a 7 pL detection volume. It is important to stress that this pH actuation principle fundamentally differs from the pH changes occurring in ionic transistors which are due to counterion enrichment and coion exclusion, because it does not rely on double-layer overlap. As a result it can be operated at high ionic strength and in channels of up to at least 1 µm height

Field-effect pH Control in Nanochannels

We demonstrate a novel capacitive method to change the pH in nanochannels.\ud The device employs metal electrodes outside an insulating channel wall to change\ud the electrical double layer potential by the field effect (‘voltage gating’). We demonstrate that this potential change is accompanied by a release or uptake of protons from surface groups, resulting in a pH shift in the nanochannel, and a titration of solution compounds present. This pH actuation method avoids redox reactions and can be very useful as a “soft” way to change the pH in small volumes e.g. in bioassays or cell-based research, but also for detection in separation methods. We demonstrate the detection of 100 attomol of TRIS in the detection volume by titration. Importantly, the proton release mechanism does not rely on double layer overlap

Electrokinetic pumping and detection of low-volume flows in nanochannels

Electrokinetic pumping of low-volume rates was performed on-chip in channels of small cross sectional area and height in the sub-m range. The flow was detected with the current monitoring technique by monitoring the change in resistance of the fluid in the channel upon the electroosmosis-driven displacement of an electrolyte solution by a second electrolyte solution. Flow rates in the order of 0.1 pL/s were successfully generated and detected. The channels were fabricated with the sacrificial layer technology

Gene transfer and protein dynamics in stem cells using single cell electroporation in a microfluidic device

There is great interest in genetic modification of bone marrow-derived mesenchymal stem cells (MSC), not only for research purposes but also for use in (autologous) patient-derived-patient-used transplantations. A major drawback of bulk methods for genetic modifications of (stem) cells, like bulk-electroporation, is its limited yield of DNA transfection (typically then 10%). This is even more limited when cells are present at very low numbers, as is the case for stem cells. Here we present an alternative technology to transfect cells with high efficiency (>75%), based on single cell electroporation in a microfluidic device. In a first experiment we show that we can successfully transport propidium iodide (PI) into single mouse myoblastic C2C12 cells. Subsequently, we show the use of this microfluidic device to perform successful electroporation of single mouse myoblastic C2C12 cells and single human MSC with vector DNA encoding a green fluorescent-erk1 fusion protein (EGFP-ERK1 (MAPK3)). Finally, we performed electroporation in combination with live imaging of protein expression and dynamics in response to extracellular stimuli, by fibroblast growth factor (FGF-2). We observed nuclear translocation of EGFP-ERK1 in both cell types within 15 min after FGF-2 stimulation. Due to the successful and promising results, we predict that microfluidic devices can be used for highly efficient small-scale genetic modification of cells, and biological experimentation, offering possibilities to study cellular processes at the single cell level. Future applications might be small-scale production of cells for therapeutic application under controlled conditions

Spatial Site-Patterning of Wettability in a Microcapillary Tube

Substrate functionalization is of great importance in successfully manipulating flows and liquid interfaces in microdevices. Herein, we propose an alternative approach for spatial patterning of wettability in a microcapillary tube. The method combines a photolithography process with self-assembled monolayer formation. The modified microcapillaries show very sharp boundaries between the alternating hydrophilic/hydrophobic segments with an achieved smallest domain dimension down to 60 μm inside a 580 μm inner diameter capillary. Our two-step method allows us to pattern multiple types of functional groups in an enclosed channel. Such structures are promising regarding the manipulation of segmented flows inside capillaries