Selective labeling of phosphatidylserine for cryo-TEM by a two-step immunogold method.

Immunogold labeling in transmission electron microscopy (TEM) utilizes the high electron density of gold nanoparticles conjugated to proteins to identify specific antigens in biological samples. In this work we applied the concept of immunogold labeling for the labeling of negatively charged phospholipids, namely phosphatidylserine, by a simple protocol, performed entirely in the liquid-phase, from which cryo-TEM specimens can be directly prepared. Labeling included a two-step process using biotinylated annexin-V and gold-conjugated streptavidin. We initially applied it on liposomal systems, demonstrating its specificity and selectivity, differentiating between 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (DOPS) membranes. We also observed specific labeling on extracellular vesicle samples isolated from THP1 cells and from MDA-468 cells, which underwent stimulations. Finally, we compared the levels of annexin-V labeling on the cells vs. on their isolated EVs by flow cytometry and found a good correlation with the cryo-TEM results. This simple, yet effective labeling technique makes it possible to differentiate between negatively charged and non-negatively charged membranes, thus shillucidating their possible EV shedding mechanism.

Tunable touch sensor and combined sensing platform: toward nanoparticle-based electronic skin.

In this paper, we present touch (or pressure) flexible sensors based on monolayer-capped nanoparticles (MCNPs) that are potentially inexpensive, could allow low-voltage operation, and could provide a platform for multifunctional applications. We show that modifying the mechanical and geometrical properties of the flexible substrates, on which the MCNP films are deposited, allows measuring a large span of loads ranging between tens of mg to tens of grams. All flexible sensors exhibited repeatable responses even after a large number of bending cycles. In addition, we show that modified platforms of those touch (or pressure) sensors allow precise detection and monitoring of environmental temperature and humidity. Relying on their superior characteristics, we were able to build an MCNP-based prototype allowing simultaneous detection and monitoring of multiple environmental parameters of touch (or pressure), humidity, and temperature. The excellent temperature (resolution higher than 1 °C and average error of ~5%) and relative humidity (resolution higher than 1% RH and average error of ~9%) sensitivities and the possibility to integrate those sensing abilities makes the suggested platform interesting for potentially inexpensive and low-voltage multifunctional electronic-skin applications.