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Both microplastics and phytoplankton are found together in the ocean as suspended microparticles. There is a need for deployable technologies that can identify, size, and count these particles at high throughput to monitor plankton community structure and microplastic pollution levels. In situ analysis is particularly desirable as it avoids the problems associated with sample storage, processing, and degradation. Current technologies for phytoplankton and microplastic analysis are limited in their capability by specificity, throughput, or lack of deployability. Little attention has been paid to the smallest size fraction of microplastics and phytoplankton below 10 µm in diameter, which are in high abundance. Impedance cytometry is a technique that uses microfluidic chips with integrated microelectrodes to measure the electrical impedance of individual particles. Here, we present an impedance cytometer that can discriminate and count microplastics sampled directly from a mixture of phytoplankton in a seawater-like medium in the 1.5-10 µm size range. A simple machine learning algorithm was used to classify microplastic particles based on dual-frequency impedance measurements of particle size (at 1 MHz) and cell internal electrical composition (at 500 MHz). The technique shows promise for marine deployment, as the chip is sensitive, rugged, and mass producible.
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Particle classification plays a crucial role in various scientific and technological applications, such as differentiating between bacteria and viruses in healthcare applications or identifying and classifying cancer cells. This technique requires accurate and efficient analysis of particle properties. In this study, we investigated the integration of electrical and optical features through a multimodal approach for particle classification. Machine learning classifier algorithms were applied to evaluate the impact of combining these measurements. Our results demonstrate the superiority of the multimodal approach over analyzing electrical or optical features independently. We achieved an average test accuracy of 94.9% by integrating both modalities, compared to 66.4% for electrical features alone and 90.7% for optical features alone. This highlights the complementary nature of electrical and optical information and its potential for enhancing classification performance. By leveraging electrical sensing and optical imaging techniques, our multimodal approach provides deeper insights into particle properties and offers a more comprehensive understanding of complex biological systems.
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Aprendizado de Máquina , Imagem Óptica , AlgoritmosRESUMO
The necessity for rapid and accurate bacterial growth monitoring is imperative across various domains, including healthcare and environmental safety. We introduce the self-synchronized droplet-amplified electrical screening cytometry (SYNC) system, a novel meld of droplet microfluidics and electrochemical amplification tailored for precise bacterial growth kinetic monitoring. SYNC encapsulates single bacteria in picolitre droplets, enabling real-time, fluorescence-free electrochemical monitoring. A specially devised phosphorylation-amplified culture medium translates bacterial metabolic activity into discernible electrical impedance changes. The dual-channel design and a rail-based structure in SYNC facilitate parallel screening and self-synchronization of droplets, addressing the limitations of conventional impedance cytometry. SYNC showcases a 5-fold enhancement in detection sensitivity and reduces 50% of the detection time compared to traditional approaches. Notably, SYNC is pioneering in providing exact initial bacterial concentrations, achieve to 104 bacteria/ml, a capability unmatched by existing real-time techniques measuring electrochemical variations. Along with its robust performance, this earmarks SYNC as a powerful tool for applications such as antibiotic susceptibility testing, food quality monitoring, and real-time water bacteria monitoring, paving the way for enhanced microbial process management and infection control.
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Técnicas Biossensoriais , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/instrumentação , Fosforilação , Desenho de Equipamento , Microfluídica/métodos , Bactérias/isolamento & purificação , Bactérias/crescimento & desenvolvimento , Cinética , Técnicas Eletroquímicas/métodos , Escherichia coliRESUMO
Microfluidic impedance cytometry (MIC) has emerged as a popular technique for single-cell analysis. Traditional MIC electrode designs consist of a pair of (or three) working electrodes, and their detection performance needs further improvements for microorganisms. In this study, we designed an 8-electrode MIC device in which the center pair was defined as the working electrode, and the connection status of bypass electrodes could be changed. This allowed us to compare the performance of layouts with no bypasses and those with floating or grounding electrodes by simulation and experiment. The results of detecting Φ 5 µm beads revealed that both the grounding and the floating electrode outperformed the no bypass electrode, and the grounding electrode demonstrated the best signal-to-noise ratio (SNR), coefficient of variation (CV), and detection sensitivity. Furthermore, the effects of different bypass grounding areas (numbers of grounding electrodes) were investigated. Finally, particles passing at high horizontal positions can be detected, and Φ 1 µm beads can be measured in a wide channel (150 µm) using a fully grounding electrode, with the sensitivity of bead volume detection reaching 0.00097%. This provides a general MIC electrode optimization technology for detecting smaller particles, even macromolecular proteins, viruses, and exosomes in the future.
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Impedância Elétrica , Eletrodos , Razão Sinal-Ruído , Microfluídica , Técnicas Biossensoriais , Desenho de Equipamento , Citometria de Fluxo , Técnicas Analíticas MicrofluídicasRESUMO
There is a paradigm shift in biomanufacturing toward continuous bioprocessing but cell-based manufacturing using adherent and suspension cultures, including microcarriers, hydrogel microparticles, and 3D cell aggregates, remains challenging due to the lack of efficient in-line bioprocess monitoring and cell harvesting tools. Herein, a novel label-free microfluidic platform for high throughput (≈50 particles/sec) impedance bioanalysis of biomass, cell viability, and stem cell differentiation at single particle resolution is reported. The device is integrated with a real-time piezo-actuated particle sorter based on user-defined multi-frequency impedance signatures. Biomass profiling of Cytodex-3 microcarriers seeded with adipose-derived mesenchymal stem cells (ADSCs) is first performed to sort well-seeded or confluent microcarriers for downstream culture or harvesting, respectively. Next, impedance-based isolation of microcarriers with osteogenic differentiated ADSCs is demonstrated, which is validated with a twofold increase of calcium content in sorted ADSCs. Impedance profiling of heterogenous ADSCs-encapsulated hydrogel (alginate) microparticles and 3D ADSC aggregate mixtures is also performed to sort particles with high biomass and cell viability to improve cell quality. Overall, the scalable microfluidic platform technology enables in-line sample processing from bioreactors directly and automated analysis of cell quality attributes to maximize cell yield and improve the control of cell quality in continuous cell-based manufacturing.
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Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Humanos , Diferenciação Celular , Sobrevivência Celular , Hidrogéis/química , Agregação Celular , Separação Celular/métodos , Alginatos/química , Tecido Adiposo/citologia , Técnicas de Cultura de Células/métodos , Técnicas de Cultura de Células/instrumentaçãoRESUMO
In this work, we demonstrate the differentiation of demodulated multifrequency signals from impedance sensitive microparticles when targeting surface receptors on neutrophils in a microfluidic impedance cytometer. These scheme uses a single signal input and detection configuration, and machine learning can differentiate particle types with up to 82% accuracy.