RESUMO
The assessment of particle and cell size in electrical microfluidic flow cytometers has become common practice. Nevertheless, in flow cytometers with coplanar electrodes accurate determination of particle size is difficult, owing to the inhomogeneous electric field. Pre-defined signal templates and compensation methods have been introduced to correct for this positional dependence, but are cumbersome when dealing with irregular signal shapes. We introduce a simple and accurate post-processing method without the use of pre-defined signal templates and compensation functions using supervised machine learning. We implemented a multiple linear regression model and show an average reduction of the particle diameter variation by 37% with respect to an earlier processing method based on a feature extraction algorithm and compensation function. Furthermore, we demonstrate its application in flow cytometry by determining the size distribution of a population of small (4.6 ± 0.9 µm) and large (5.9 ± 0.8 µm) yeast cells. The improved performance of this coplanar, two electrode chip enables precise cell size determination in easy to fabricate impedance flow cytometers.
Assuntos
Técnicas Analíticas Microfluídicas , Microfluídica , Microfluídica/métodos , Citometria de Fluxo/métodos , Impedância Elétrica , Tamanho da Partícula , Aprendizado de Máquina Supervisionado , Técnicas Analíticas Microfluídicas/métodosRESUMO
Endotoxin is a deadly pyrogen, rendering it crucial to monitor with high accuracy and efficiency. However, current endotoxin detection relies on multistep processes that are labor-intensive, time-consuming, and unsustainable. Here, we report an aptamer-based biosensor for the real-time optical detection of endotoxin. The endotoxin sensor exploits the distance-dependent scattering of gold nanoparticles (AuNPs) coupled to a gold nanofilm. This is enabled by the conformational changes of an endotoxin-specific aptamer upon target binding. The sensor can be used in an ensemble mode and single-particle mode under dark-field illumination. In the ensemble mode, the sensor is coupled with a microspectrometer and exhibits high specificity, reliability (i.e., linear concentration to signal profile in logarithmic scale), and reusability for repeated endotoxin measurements. Individual endotoxins can be detected by monitoring the color of single AuNPs via a color camera, achieving single-molecule resolution. This platform can potentially advance endotoxin detection to safeguard medical, food, and pharmaceutical products.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Endotoxinas , Ouro/química , Reprodutibilidade dos Testes , Aptâmeros de Nucleotídeos/química , Nanopartículas Metálicas/química , Limite de DetecçãoRESUMO
Lab-on-chip electrokinetic focusing and separation techniques are widely used in several scientific fields. In a number of cases, these techniques have been combined with a selective analyte extraction for off-chip analysis. Nevertheless, the usability of the extracts is limited by diffusion which reduces the separation resolution. In this paper we propose the integration of a droplet generator capable of continuous or on-demand generation and extraction of electrokinetically separated and focused analytes. We demonstrate the selective droplet extraction of model analytes separated and concentrated via ion concentration polarization focusing (ICPF). We report extracted droplets with 1000-fold increased concentration. Importantly, the droplet generator does not interrupt the ICPF process making it suitable for integration with the majority of electrokinetic separation techniques.
RESUMO
Electrokinetic separation techniques in microfluidics are a powerful analytical chemistry tool, although an inherent limitation of microfluidics is their low sample throughput. In this article we report a free-flow variant of an electrokinetic focusing method, namely ion concentration polarization focusing (ICPF). The analytes flow continuously through the system via pressure driven flow while they separate and concentrate perpendicularly to the flow by ICPF. We demonstrate free flow ion concentration polarization focusing (FF-ICPF) in two operating modes, namely peak and plateau modes. Additionally, we showed the separation resolution could be improved by the use of an electrophoretic spacer. We report a concentration factor of 10 in human blood plasma in continuous flow at a flow rate of 15 µL min-1.
RESUMO
Electrokinetic focusing and separation methods, specifically ion concentration polarization focusing (ICPF), provide a very powerful and easy to use analytical tool for several scientific fields. Nevertheless, the concentrated and separated analytes are effectively trapped inside the chip in picoliter volumes. In this article we propose an ICPF device that allows continuous and selective extraction of the focused analytes. A theoretical background is presented to understand the dynamics of the system and a 1D model was developed that describes the general behavior of the system. We demonstrate the selective extraction of three fluorescent model anionic analytes and we report selective extraction of the analytes at a 300-fold increased concentration.
RESUMO
Microparticle porosity is normally determined in bulk manner providing an ensemble average that hinders establishing the individual role of each microparticle. On the other hand, single particle characterization implies expensive technology. We propose to use ion concentration polarization to measure differences in mesoporosity at the single particle level. Ion concentration polarization occurs at the interface between an electrolyte and a porous particle when an electric field is applied. The extent of ion concentration polarization depends, among others, on the mesopore size and density. By using a fluorescence marker, we could measure differences in concentration polarization between particles with 3 and 13 nm average mesopore diameters. A qualitative model was developed in order to understand and interpret the phenomena. We believe that this inexpensive method could be used to measure differences in mesoporous particle materials such as catalysts.