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Fluorescent sensory MIP (molecularly imprinted polymer) particles were combined with a droplet-based 3D microfluidic system for the sensitive and selective determination of 2,4-dichlorophenoxyacetic acid (2,4-D) in water samples. 2,4-D being an important and widely used herbicide to regulate plant growth, its extensive use leads to food or ground water contamination. Analytical assays based on MIP have emerged as a valuable tool in the field of environmental analysis thanks to low production costs, stability, format adaptability and their ability to recognize a wide variety of targets, such as the critical neutral organic molecules present in water: herbicides, pesticides, antibiotics, etc.
To accomplish this, a tailor-made fluorescent indicator cross-linker was designed that translates directly the binding event into an enhanced fluorescence signal. This phenoxazinone-type cross-linker was co-polymerized into a thin MIP layer grafted from the surface of silica microparticles. The latter was achieved by integration of the fluorescent core-shell MIP sensor particles into a modular microfluidic platform that allows for an in-line phase-transfer assay, extracting the analyte from aqueous sample droplets into organic phase droplets that contain the sensor particles.
This tool offering a novel simple and rapid way for the detection of herbicides, real-time fluorescence determination of 2,4-D down to 20 nM was realized with the system and applied for the analysis of worlwide surface water samples.
The identification and quantification of phosphorylated amino acids, peptides and proteins is one of the key challenges in contemporary bioanalytical research, noteworthy to diagnose and treat diseases at an early stage. Small sensing devices like microfluidic chips combined with “smart” detection chemistry and simple data assessment, processing and presentation are attractive in this regard. We developed novel synthetic probes, targeting phosphorylated amino acids, based on core-shell microparticles consisting of a silica core coated with a molecularly imprinted polymer shell (MIP). These “plastic antibodies” which are extremely robust, resist denaturing solvents and high temperatures, can be reproducibly produced at low cost, can potentially overcome many of the problems in the current detection strategies. The MIP layer, containing a fluorescent probe monomer, responds to phosphorylated amino acids such as phosphorylated tyrosine (pTyr) with a significant imprinting factor, i.e. selectivity, higher than 3.5, and a “lighting-up” of its fluorescence accompanied by the development of a strongly red-shifted emission band. The bead-based ratiometric detection scheme has also been successfully transferred to a microfluidic chip format to demonstrate its applicability to rapid assays. Such a device could yield an automated pTyr measurement system in the future. The setup was built by coupling a PDMS/Teflon/glass microfluidic chip with an optical setup for fluorescence measurements able to extract and monitor pTyr concentration from 0.5–200 μM.
Vesicles self-assembled in water from natural and synthetic phospholipids (liposomes), amphiphilic block copolymers (polymersomes), and more recently amphiphilic Janus dendrimers (dendrimersomes) -5 as hollow soft structures in the nano size regime have attracted increasing interest as they can mimic primitive and contemporary biological membranes, and can be configured into biomimetic nanocapsules with application in nanomedicine such as gene, proteins and drug carriers or theranostics.
Compared to other amphiphilic structures, the molecular structure of Janus dendrimers can be precisely controlled: by using the vast range of tools from organic chemistry their size, architecture, density, generation as well as the number of end groups of the individual dendrons can be modified as desired.
Unfortunately, the controlled production of supramolecular aggregates made thereof is still a challenging task. Conventional batch-based techniques such as the solvent injection method or the film hydration method typically go along with a lack of control over self assembly/mixing and thus over size, morphology and size distribution of the vesicles.
The micromixer technology is a promising method for the controlled preparation of supramolecular assemblies as it allows control of mixing at microscale level. In addition, such microfluidic systems benefit from a high mixing efficiency, a low mixing time as well as from a reproducible and continuous production of soft nanoparticles. Herein, we report on the microfluidic-controlled self-assembly of Janus dendrimers as dendrimersomes and the impact of the mixing parameters on the self-assembly process.
Chlorination of pool water and wastewater, in food and pharmaceutical production, as well as in pesticide and paper manufacturing is a routinely used technique. However, the amount of chlorine in water must be strictly adjusted, to ensure enough concentration to kill pathogenic bacteria and viruses, while preventing too high concentrations inducing negative effects on human health. As an indicator, a molecular fluorescent probe based on a BODIPY structure was designed. This indicator exhibits a sensitive and selective fluorescence response upon increasing concentrations of hypochlorite in aqueous solvent mixtures. Real-time analyses became possible after the integration of this fluorescent indicator into newly designed 2D & 3D microfluidic chips incorporating a passive sinusoidal mixer and a micro-hydrocyclone, respectively. A comparison of the two microfluidic systems, including their ability to prevent accumulation or circulation of microbubbles, has shown excellent fluidic behaviour for the micro-hydrocyclone device. This system was distinctly more robust against gas bubbles, showed a higher signal gain and allowed to halve the limit of detection to 0.02 mg L–1. The use of the 3D system to quantify the chlorine content of pool water samples for sensitive and quantitative chlorine monitoring has been demonstrated.
This study describes the motion of magnetizable particles of spherical shape with radius at most of 1 μm, also referred to as microbeads, immersed in a liquid under laminar flow conditions in a microfluidic channel. An external magnetic field is applied in part of the system such that it permeates the channel and is characterized by a spatially varying magnetic field, i.e., the gradient of the magnetic field is nonzero. The beads are superparamagnetic; hence they can reach high level of magnetization in the presence of a magnetic field, and this magnetization disappears when the field is zero. Therein lies the attractiveness of these microbeads and the potential for applications because their motion can be controlled using an external magnetic field. The motion is governed by several factors, including the magnetic force acting on microbeads (particles), the drag force due to viscosity, the interaction between particles and the fluid, as well as the interaction between particles themselves. For a single particle case, the trajectory of the motion is determined by balancing the drag and magnetic forces acting on the particle, a calculation that in general requires numerical integration. For a system consisting of an ensemble of interacting particles, several regimes of motion dominated by one of the factors are identified. Of particular interest are the systems dominated by the large number of particles in the ensemble, i.e., high particle concentration, in which cases the wake influences the flow downstream. This effect is qualitatively investigated by considering the Navier-Stokes equation with and without the magnetic force contribution.
pH and oxygen are amongst the most important and frequently measured analytes in the life and material sciences, indicating, e.g., diseases and corrosion processes. This includes the optical monitoring of pH in living cells for studying cellular internalization pathways, such as phagocytosis, endocytosis, and receptor ligand internalization with the aid of molecular and nanoscale fluorescent sensors. Nanoparticle (NP)-based sensors, that are labeled or stained with a multitude of sensor dyes, have several advantages as compare to conventional molecular probes like enhanced brightness, i.e., amplified signals, ease of designing ratiometric systems by combining analyte sensitive and inert reference dyes, and increased photostability. Moreover, this can enable the use of hydrophobic dyes in aqueous environments. Versatile templates and carriers for the fabrication of nanosensors by the staining and/or labelling with different fluorophores and sensor molecules or surface functionalized NP like silica (SiO2-NP) and polystyrene (PS-NP) particles provide. Here we present the design of a versatile platform of color emissive nanosensors and stimuli-responsive microparticles for the measurement of pH, oxygen, and other targets utilizing both types of matrices and sets of spectrally distinguishable sensor and reference dyes and their characterization and demonstrate the applicability of representative sensor particle for cellular studies.
PFAS Sensors
(2023)
This contribution provides an introduction to the development of sensors for PFAS analysis, presents the most common approaches, and describes the opto-microfluidic strategy in combination with polymerizable indicators and detection matrices currently being pursued by the Chemical and Optical Sensing Division at BAM.
Per- and polyfluoroalkyl substances (PFAS) are a class of man-made organo-fluorine chemicals that have become environmental contaminants of emerging concern, originating from a variety of materials such as adhesive, stain- and oil-resistant coatings, firefighting foams, etc. The high strength of this C-F bond makes PFAS thermodynamically stable and resistant to (bio)degradation, thus retaining them in the environment over time. Perfluoroalkyl carboxylic acids (PFCAs), one category of the most used PFAS, consist of a fully fluorinated carbon backbone and a charged carboxylic acid headgroup, and have been classified as Substances of Very High Concern (SVHC) and added to the REACH Candidate List due to their persistence in the environment, non-biodegradability and toxicological effects.[1-2] Traditional techniques for the analysis of PFCAs include GC-MS, HRMS and HPLC-based approaches, which are laborious, not portable, costly and require trained personnel. In contrast, fluorescence assays can be designed as easy-to-operate, portable and cost-effective methods with high sensitivity and fast response. Integration of fluorescent probes with an adequately miniaturized assay enables a promising alternative for PFCAs analysis.
Here, a novel guanidine fluorescent probe has been synthesized and fully characterized for the detection of PFCAs in a biphasic extract-&-detect assay. The fluorescent probe was then incorporated into polymeric matrices supported by a red dye-doped SiO2 nanoparticle to construct a dual-emission sensing platform. Such a system allows precise and selective detection of PFCAs, reducing the interference of competitors, matrix effects and other factors except for the PFCAs. The system was then employed in a droplet-based microfluidic setup which offers a portable and easy to operate detection platform.