Real time imaging of single extracellular vesicle pH regulation in a microfluidic cross-flow filtration platform

Extracellular vesicles (EVs) are cell-derived membranous structures carrying transmembrane proteins and luminal cargo. Their complex cargo requires pH stability in EVs while traversing diverse body fluids. We used a filtration-based platform to capture and stabilize EVs based on their size and studied their pH regulation at the single EV level. Dead-end filtration facilitated EV capture in the pores of an ultrathin (100 nm thick) and nanoporous silicon nitride (NPN) membrane within a custom microfluidic device. Immobilized EVs were rapidly exposed to test solution changes driven across the backside of the membrane using tangential flow without exposing the EVs to fluid shear forces. The epithelial sodium-hydrogen exchanger, NHE1, is a ubiquitous plasma membrane protein tasked with the maintenance of cytoplasmic pH at neutrality. We show that NHE1 identified on the membrane of EVs is functional in the maintenance of pH neutrality within single vesicles. This is the first mechanistic description of EV function on the single vesicle level.

Laser-excited elastic guided waves reveal the complex mechanics of nanoporous silicon

Nanoporosity in silicon leads to completely new functionalities of this mainstream semiconductor. A difficult to assess mechanics has however significantly limited its application in fields ranging from nanofluidics and biosensorics to drug delivery, energy storage and photonics. Here, we present a study on laser-excited elastic guided waves detected contactless and non-destructively in dry and liquid-infused single-crystalline porous silicon. These experiments reveal that the self-organised formation of 100 billions of parallel nanopores per square centimetre cross section results in a nearly isotropic elasticity perpendicular to the pore axes and an 80% effective stiffness reduction, altogether leading to significant deviations from the cubic anisotropy observed in bulk silicon. Our thorough assessment of the wafer-scale mechanics of nanoporous silicon provides the base for predictive applications in robust on-chip devices and evidences that recent breakthroughs in laser ultrasonics open up entirely new frontiers for in-situ, non-destructive mechanical characterisation of dry and liquid-functionalised porous materials.