Proteome Analysis of Ground State Pluripotency.

The differentiation potential of pluripotent embryonic stem cells (ESCs) can be manipulated via serum and medium conditions for direct cellular development or to maintain a naïve ground state. The self-renewal state of ESCs can thus be induced by adding inhibitors of mitogen activated protein kinase (MAPK) and glycogen synthase kinase-3 (Gsk3), known as 2 inhibitors (2i) treatment. We have used a shotgun proteomics approach to investigate differences in protein expressions between 2i- and serum-grown mESCs.

High-Precision Pinpointing of Luminescent Targets in Encoder-Assisted Scanning Microscopy Allowing High-Speed Quantitative Analysis.

Compared with routine microscopy imaging of a few analytes at a time, rapid scanning through the whole sample area of a microscope slide to locate every single target object offers many advantages in terms of simplicity, speed, throughput, and potential for robust quantitative analysis. Existing techniques that accommodate solid-phase samples incorporating individual micrometer-sized targets generally rely on digital microscopy and image analysis, with intrinsically low throughput and reliability. Here, we report an advanced on-the-fly stage scanning method to achieve high-precision target location across the whole slide.

A novel biocompatible europium ligand for sensitive time-gated immunodetection.

We describe the synthesis of a novel hydrophilic derivative of a tetradentate β-diketone europium ligand that was used to prepare an immunoconjugate probe against Giardia lamblia cysts. We used a Gated Autosynchronous Luminescence Detector (GALD) to obtain high quality delayed luminescence images of cells 30-fold faster than ever previously reported.

Surface Plasmon Scattering in Exposed Core Optical Fiber for Enhanced Resolution Refractive Index Sensing.

Refractometric sensors based on optical excitation of surface plasmons on the side of an optical fiber is an established sensing architecture that has enabled laboratory demonstrations of cost effective portable devices for biological and chemical applications. Here we report a Surface Plasmon Resonance (SPR) configuration realized in an Exposed Core Microstructured Optical Fiber (ECF) capable of optimizing both sensitivity and resolution. To the best of our knowledge, this is the first demonstration of fabrication of a rough metal coating suitable for spectral interrogation of scattered plasmonic wave using chemical electroless plating technique on a 10 μm diameter exposed core of the ECF.

Background-free in-vivo Imaging of Vitamin C using Time-gateable Responsive Probe.

Sensitive optical imaging of active biomolecules in the living organism requires both a molecular probe specifically responsive to the target and a high-contrast approach to remove the background interference from autofluorescence and light scatterings. Here, a responsive probe for ascorbic acid (vitamin C) has been developed by conjugating two nitroxide radicals with a long-lived luminescent europium complex. The nitroxide radical withholds the probe on its "off" state (barely luminescent), until the presence of vitamin C will switch on the probe by forming its hydroxylamine derivative.

Photoswitchable membranes based on peptide-modified nanoporous anodic alumina: toward smart membranes for on-demand molecular transport.

A smart and reversibly photoswitchable membrane based on an azobenzene photo-switch containing peptides attached inside the pores of nanoporous anodic alumina membranes (NAAMs) is presented. The transport of molecules of interest across the photoswitchable peptide (PSP) functionalized NAAMs can be effectively controlled and manipulated as a function of the photostationary state of the azobenzene group in a PSP.

The correlation of electrochemical measurements and molecular junction conductance simulations in β-strand peptides.

Understanding the electronic properties of single peptides is not only of fundamental importance, but it is also paramount to the realization of peptide-based molecular electronic components. Electrochemical and theoretical studies are reported on two β-strand-based peptides, one with its backbone constrained with a triazole-containing tether introduced by Huisgen cycloaddition (peptide 1) and the other a direct linear analogue (peptide 2). Density functional theory (DFT) and non-equilibrium Green's function were used to investigate conductance in molecular junctions containing peptides 3 and 4 (analogues of 1 and 2).

A fiber-tip label-free biological sensing platform: a practical approach toward in-vivo sensing.

The platform presented here was devised to address the unmet need for real time label-free in vivo sensing by bringing together a refractive index transduction mechanism based on Whispering Gallery Modes (WGM) in dye doped microspheres and Microstructured Optical Fibers. In addition to providing remote excitation and collection of the WGM signal, the fiber provides significant practical advantages such as an easy manipulation of the microresonator and the use of this sensor in a dip sensing architecture, alleviating the need for a complex microfluidic interface. Here, we present the first demonstration of the use of this approach for biological sensing and evaluate its limitation in a sensing configuration deprived of liquid flow which is most likely to occur in an in vivo setting.

Practical implementation, characterization and applications of a multi-colour time-gated luminescence microscope.

Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived complexes under low-background conditions. This is realized by imaging two pathogenic organisms (Giardia lamblia stained with a red europium probe and Cryptosporidium parvum with a green terbium probe) at UV wavelengths (320-400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit.

Unraveling the interplay of backbone rigidity and electron rich side-chains on electron transfer in peptides: the realization of tunable molecular wires.

Electrochemical studies are reported on a series of peptides constrained into either a 310-helix (1-6) or β-strand (7-9) conformation, with variable numbers of electron rich alkene containing side chains. Peptides (1 and 2) and (7 and 8) are further constrained into these geometries with a suitable side chain tether introduced by ring closing metathesis (RCM). Peptides 1, 4 and 5, each containing a single alkene side chain reveal a direct link between backbone rigidity and electron transfer, in isolation from any effects due to the electronic properties of the electron rich side-chains.

On-the-fly decoding luminescence lifetimes in the microsecond region for lanthanide-encoded suspension arrays.

Significant multiplexing capacity of optical time-domain coding has been recently demonstrated by tuning luminescence lifetimes of the upconversion nanoparticles called 'τ-Dots'. It provides a large dynamic range of lifetimes from microseconds to milliseconds, which allows creating large libraries of nanotags/microcarriers. However, a robust approach is required to rapidly and accurately measure the luminescence lifetimes from the relatively slow-decaying signals.