-
Frontiers in Bioengineering and... 2024
PubMed: 38895556
DOI: 10.3389/fbioe.2024.1432352 -
APL Bioengineering Jun 2024Micropipette aspiration (MPA) is one of the gold standards for quantifying biological samples' mechanical properties, which are crucial from the cell membrane scale to...
Micropipette aspiration (MPA) is one of the gold standards for quantifying biological samples' mechanical properties, which are crucial from the cell membrane scale to the multicellular tissue. However, relying on the manipulation of individual home-made glass pipettes, MPA suffers from low throughput and no automation. Here, we introduce the sliding insert micropipette aspiration method, which permits parallelization and automation, thanks to the insertion of tubular pipettes, obtained by photolithography, within microfluidic channels. We show its application both at the lipid bilayer level, by probing vesicles to measure membrane bending and stretching moduli, and at the tissue level by quantifying the viscoelasticity of 3D cell aggregates. This approach opens the way to high-throughput, quantitative mechanical testing of many types of biological samples, from vesicles and individual cells to cell aggregates and explants, under dynamic physico-chemical stimuli.
PubMed: 38894959
DOI: 10.1063/5.0193333 -
Sensors (Basel, Switzerland) Jun 2024Fabry disease is a lysosomal storage disorder caused by a significant decrease in the activity or absence of the enzyme α-galactosidase A. The diagnostics of Fabry...
Fabry disease is a lysosomal storage disorder caused by a significant decrease in the activity or absence of the enzyme α-galactosidase A. The diagnostics of Fabry disease during newborn screening are reasonable, due to the availability of enzyme replacement therapy. This paper presents an electrochemical method using complementary metal-oxide semiconductor (CMOS)-compatible ion-sensitive field effect transistors (ISFETs) with hafnium oxide-sensitive surfaces for the detection of α-galactosidase A activity in dried blood spot extracts. The capability of ISFETs to detect the reaction catalyzed by α-galactosidase A was demonstrated. The buffer composition was optimized to provide suitable conditions for both enzyme and ISFET performance. The use of ISFET structures as sensor elements allowed for the label-free detection of enzymatic reactions with melibiose, a natural substrate of α-galactosidase A, instead of a synthetic fluorogenic one. ISFET chips were packaged with printed circuit boards and microfluidic reaction chambers to enable long-term signal measurement using a custom device. The packaged sensors were demonstrated to discriminate between normal and inhibited GLA activity in dried blood spots extracts. The described method offers a promising solution for increasing the widespread distribution of newborn screening of Fabry disease.
Topics: alpha-Galactosidase; Dried Blood Spot Testing; Humans; Fabry Disease; Transistors, Electronic; Biosensing Techniques; Infant, Newborn; Neonatal Screening
PubMed: 38894470
DOI: 10.3390/s24113681 -
Sensors (Basel, Switzerland) Jun 2024Hydrogels are of great importance for functionalizing sensors and microfluidics, and poly(ethylene glycol) dimethacrylate (PEG-DMA) is often used as a viscosifier for...
Hydrogels are of great importance for functionalizing sensors and microfluidics, and poly(ethylene glycol) dimethacrylate (PEG-DMA) is often used as a viscosifier for printable hydrogel precursor inks. In this study, 1-10 kDa PEG-DMA based hydrogels were characterized by gravimetric and electrochemical methods to investigate the diffusivity of small molecules and proteins. Swelling ratios () of 14.43-9.24, as well as mesh sizes ξ of 3.58-6.91 nm were calculated, and it was found that the correlates with the molar concentration of PEG-DMA in the ink () (SR = 0.1127 × MCI + 8.3256, R = 0.9692) and ξ correlates with the molecular weight () (ξ = 0.3382 × + 3.638, R = 0.9451). To investigate the sensing properties, methylene blue (MB) and MB-conjugated proteins were measured on electrochemical sensors with and without hydrogel coating. It was found that on sensors with 10 kDa PEG-DMA hydrogel modification, the DPV peak currents were reduced to 92 % for MB, 73 % for MB-BSA, and 23 % for MB-IgG. To investigate the diffusion properties of MB(-conjugates) in hydrogels with 1-10 kDa PEG-DMA, diffusivity was calculated from the current equation. It was found that diffusivity increases with increasing ξ. Finally, the release of MB-BSA was detected after drying the MB-BSA-containing hydrogel, which is a promising result for the development of hydrogel-based reagent reservoirs for biosensing.
PubMed: 38894467
DOI: 10.3390/s24113678 -
Sensors (Basel, Switzerland) May 2024Circulating tumor cells are typically found in the peripheral blood of patients, offering a crucial pathway for the early diagnosis and prediction of cancer. Traditional...
Circulating tumor cells are typically found in the peripheral blood of patients, offering a crucial pathway for the early diagnosis and prediction of cancer. Traditional methods for early cancer diagnosis are inefficient and inaccurate, making it difficult to isolate tumor cells from a large number of cells. In this paper, a new spiral microfluidic chip with asymmetric cross-section is proposed for rapid, high-throughput, label-free enrichment of CTCs in peripheral blood. A mold of the desired flow channel structure was prepared and inverted to make a trapezoidal cross-section using a micro-nanotechnology process of 3D printing. After a systematic study of how flow rate, channel width, and particle concentration affect the performance of the device, we utilized the device to simulate cell sorting of 6 μm, 15 μm, and 25 μm PS (Polystyrene) particles, and the separation efficiency and separation purity of 25 μm PS particles reached 98.3% and 96.4%. On this basis, we realize the enrichment of a large number of CTCs in diluted whole blood (5 mL). The results show that the separation efficiency of A549 was 88.9% and the separation purity was 96.4% at a high throughput of 1400 μL/min. In conclusion, we believe that the developed method is relevant for efficient recovery from whole blood and beneficial for future automated clinical analysis.
Topics: Humans; Cell Separation; Neoplastic Cells, Circulating; Lab-On-A-Chip Devices; A549 Cells; Microfluidic Analytical Techniques; Printing, Three-Dimensional
PubMed: 38894343
DOI: 10.3390/s24113552 -
Sensors (Basel, Switzerland) May 2024With the steady increase in allergy prevalence worldwide, there is a strong need for novel diagnostic tools for precise, fast, and less invasive testing methods. Herein,...
With the steady increase in allergy prevalence worldwide, there is a strong need for novel diagnostic tools for precise, fast, and less invasive testing methods. Herein, a miniatured fluorescence-based biosensing system is developed for the rapid and quantitative detection of allergen-specific immunoglobulin-E. An antibody-based fluorescence assay in a microfluidic-patterned slide, combined with a custom-made portable fluorescence reader for image acquisition and user-friendly software for the data analysis, enables obtaining results for multiple allergens in just ~1 h with only 80 μL of blood serum. The multiplexed detection of common birch, timothy grass, cat epithelia, house dust mite, and dog epithelia shows quantitative IgE-mediated allergic responses to specific allergens in control serum samples with known total IgE concentration. The responses are verified with different control tests and measurements with a commercial fluorescence reader. These results open the door to point-of-care allergy screening for early diagnosis and broader access and for large-scale research in allergies.
Topics: Biosensing Techniques; Allergens; Point-of-Care Systems; Immunoglobulin E; Animals; Humans; Hypersensitivity; Fluorescence; Dogs; Cats
PubMed: 38894075
DOI: 10.3390/s24113280 -
Molecules (Basel, Switzerland) May 2024Within the fields of infectious disease diagnostics, microfluidic-based integrated technology systems have become a vital technology in enhancing the rapidity, accuracy,... (Review)
Review
Within the fields of infectious disease diagnostics, microfluidic-based integrated technology systems have become a vital technology in enhancing the rapidity, accuracy, and portability of pathogen detection. These systems synergize microfluidic techniques with advanced molecular biology methods, including reverse transcription polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), and clustered regularly interspaced short palindromic repeats (CRISPR), have been successfully used to identify a diverse array of pathogens, including COVID-19, Ebola, Zika, and dengue fever. This review outlines the advances in pathogen detection, attributing them to the integration of microfluidic technology with traditional molecular biology methods and smartphone- and paper-based diagnostic assays. The cutting-edge diagnostic technologies are of critical importance for disease prevention and epidemic surveillance. Looking ahead, research is expected to focus on increasing detection sensitivity, streamlining testing processes, reducing costs, and enhancing the capability for remote data sharing. These improvements aim to achieve broader coverage and quicker response mechanisms, thereby constructing a more robust defense for global public health security.
Topics: Humans; Nucleic Acid Amplification Techniques; Molecular Diagnostic Techniques; Microfluidics; Communicable Diseases; COVID-19; SARS-CoV-2; Microfluidic Analytical Techniques; Dengue; Zika Virus Infection; Zika Virus
PubMed: 38893293
DOI: 10.3390/molecules29112417 -
International Journal of Molecular... May 2024Microbial foodborne pathogens present significant challenges to public health and the food industry, requiring rapid and accurate detection methods to prevent infections... (Review)
Review
Microbial foodborne pathogens present significant challenges to public health and the food industry, requiring rapid and accurate detection methods to prevent infections and ensure food safety. Conventional single biosensing techniques often exhibit limitations in terms of sensitivity, specificity, and rapidity. In response, there has been a growing interest in multimodal biosensing approaches that combine multiple sensing techniques to enhance the efficacy, accuracy, and precision in detecting these pathogens. This review investigates the current state of multimodal biosensing technologies and their potential applications within the food industry. Various multimodal biosensing platforms, such as opto-electrochemical, optical nanomaterial, multiple nanomaterial-based systems, hybrid biosensing microfluidics, and microfabrication techniques are discussed. The review provides an in-depth analysis of the advantages, challenges, and future prospects of multimodal biosensing for foodborne pathogens, emphasizing its transformative potential for food safety and public health. This comprehensive analysis aims to contribute to the development of innovative strategies for combating foodborne infections and ensuring the reliability of the global food supply chain.
Topics: Biosensing Techniques; Foodborne Diseases; Food Microbiology; Humans; Food Safety
PubMed: 38892147
DOI: 10.3390/ijms25115959 -
Cells May 2024The fungi is found in cereals and feedstuffs and may produce mycotoxins, which are secondary metabolites, such as the T-2 toxin (T-2). In this work, we explored the...
The fungi is found in cereals and feedstuffs and may produce mycotoxins, which are secondary metabolites, such as the T-2 toxin (T-2). In this work, we explored the hepatotoxicity of T-2 using microfluidic 3D hepatic cultures. The objectives were: (i) exploring the benefits of microfluidic 3D cultures compared to conventional 3D cultures available commercially (Aggrewell plates), (ii) establishing 3D co-cultures of hepatic cells (HepG2) and stellate cells (LX2) and assessing T-2 exposure in this model, (iii) characterizing the induction of metabolizing enzymes, and (iv) evaluating inflammatory markers upon T-2 exposure in microfluidic hepatic cultures. Our results demonstrated that, in comparison to commercial (large-volume) 3D cultures, spheroids formed faster and were more functional in microfluidic devices. The viability and hepatic function decreased with increasing T-2 concentrations in both monoculture and co-cultures. The RT-PCR analysis revealed that exposure to T-2 upregulates the expression of multiple Phase I and Phase II hepatic enzymes. In addition, several pro- and anti-inflammatory proteins were increased in co-cultures after exposure to T-2.
Topics: T-2 Toxin; Humans; Hep G2 Cells; Spheroids, Cellular; Liver; Coculture Techniques; Microfluidics; Hepatocytes; Cell Survival
PubMed: 38891032
DOI: 10.3390/cells13110900 -
Foods (Basel, Switzerland) May 2024Microfluidic technology, as a continuous and mass preparation method of nanoparticles, has attracted much attention in recent years. In this study, zein nanoparticles...
Microfluidic technology, as a continuous and mass preparation method of nanoparticles, has attracted much attention in recent years. In this study, zein nanoparticles (ZNPs) were continuously fabricated in a highly controlled manner by combining a microfluidics platform with the antisolvent method. The impact of ethanol content (60~95%, /) and flow rates of inner and outer phases in the microfluidics platform on particle properties were examined. Among all ZNPS, 90%-ZNPs have the highest solubility (32.83%) and the lowest hydrophobicity (90.43), which is the reverse point of the hydrophobicity of ZNPs. Moreover, when the inner phase flow rate was 1.5 mL/h, the particle size decreased significantly from 182.81 nm to 133.13 nm as the outer phase flow rate increased from 10 mL/h to 50 mL/h. The results revealed that ethanol content had significant impacts on hydrophilic-hydrophobic properties of ZNPs. The flow rates of ethanol-water solutions and deionized water (solvent and antisolvent) in the microfluidics platform significantly affected the particle size of ZNPs. These findings demonstrated that the combined application of a microfluidics platform and an antisolvent method could be an effective pathway for precisely controlling the fabrication process of protein nanoparticles and modulating their physicochemical properties.
PubMed: 38890958
DOI: 10.3390/foods13111730