Small extracellular vesicles (sEVs) generated from the endolysosomal system, often referred to as exosomes, have attracted interest as a suitable biomarker for cancer diagnostics, as they carry valuable biological information and reflect their cells of origin. Herein, we propose a simple and inexpensive electrical method for label-free detection and profiling of sEVs in the size range of exosomes. The detection method is based on the electrokinetic principle, where the change in the streaming current is monitored as the surface markers of the sEVs interact with the affinity reagents immobilized on the inner surface of a silica microcapillary. As a proof-of-concept, we detected sEVs derived from the non-small-cell lung cancer (NSCLC) cell line H1975 for a set of representative surface markers, such as epidermal growth factor receptor (EGFR), CD9, and CD63. The detection sensitivity was estimated to be ∼175000 sEVs, which represents a sensor surface coverage of only 0.04%. We further validated the ability of the sensor to measure the expression level of a membrane protein by using sEVs displaying artificially altered expressions of EGFR and CD63, which were derived from NSCLC and human embryonic kidney (HEK) 293T cells, respectively. The analysis revealed that the changes in EGFR and CD63 expressions in sEVs can be detected with a sensitivity in the order of 10% and 3%, respectively, of their parental cell expressions. The method can be easily parallelized and combined with existing microfluidic-based EV isolation technologies, allowing for rapid detection and monitoring of sEVs for cancer diagnosis.

In this paper, we present a novel approach to enhance the sensitivity of microfluidic biosensor platforms with self-assembled magnetic bead chains. An adjustable, more than 5-fold sensitivity enhancement is achieved by introducing a magnetic field gradient along a microfluidic channel by means of a soft-magnetic lattice with a 350 μm spacing. The alternating magnetic field induces the self-assembly of the magnetic beads in chains or clusters and thus improves the perfusion and active contact between the analyte and the beads. The soft-magnetic lattices can be applied independent of the channel geometry or chip material to any microfluidic biosensing platform. At the same time, the bead-based approach achieves chip reusability and shortened measurement times. The bead chain properties and the maximum flow velocity for bead retention were validated by optical microscopy in a glass capillary. The magnetic actuation system was successfully validated with a biotin-streptavidin model assay on a low-cost electrochemical microfluidic chip, fabricated by dry-film photoresist technology (DFR). Labelling with glucose oxidase (GOx) permits rapid electrochemical detection of enzymatically produced H2O2.

No abstract

A recently presented novel plasma source generating discharge in liquids based on the pin-hole discharge configuration is characterized in detail. The system is supplied by DC non-pulsing high voltage of both polarities in NaCl water solutions at a conductivity range of 100-15 000 μS/cm. The discharge itself shows self-pulsing operation. The discharge ignition is observed in micro bubbles by transient discharge followed by a glow discharge in positive polarity at lower conductivities propagating inside the bubbles. At high conductivities, the glow regime is particularly replaced by a more energetic sequence of transient discharges followed by a shorter glow mode operation. The transient regime probability and its intensity are higher in the negative discharge polarity. The transient discharge produces acoustic waves and shock waves, which are observed at the moment of the bubble cavitation. The average gas temperature of 700-1500 K was calculated from the lowest OH (A-X) 0-0 band transitions. The average electron concentrations of 10 20 -10 23 m −3 were calculated from H α and H β line profiles. Finally, the production of a chemically active species is determined by hydrogen peroxide energy yields related to the energy consumption of the whole interelectrode system. All these quantities are dependent on the solution conductivity, the discharge polarity, and the applied power.