Novel Reproducible Manufacturing and Reversible Sealing Method for Microfluidic Devices

Pérez-Sosa, Camilo; Peñaherrera, Ana; Rosero, Gustavo; Bourguignon, Natalia; Aravelli, Aparna; Bhansali, Shekhar; Pérez, Maximiliano; Lerner, Betiana

doi:10.3390/mi13050650

Cited by 8 publications

(6 citation statements)

References 18 publications

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“…A great number of microdevices are suitable for single use only, and it is difficult to recover the contents inside the microchannels or perform advanced microscopy visualization due to their irreversible sealing method. On the flip side, conventional reversible sealing procedures are only suitable for low pressures (less than 35 kPa), and chips are prone to debonding or leaking [ 62 ]. Irreversible sealing methodologies are useful due to their strong bonding and ability to support high pressures.…”

Section: Resultsmentioning

confidence: 99%

Demonstration of a Transparent and Adhesive Sealing Top for Microfluidic Lab-Chip Applications

Agarwal,

Salahuddin,

Ahamed

2024

Sensors

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A transparent and adhesive film-based enclosing and sealing method is here presented for out-of-cleanroom-based open-form microfluidic devices. The commercially available polyester flexible film known as Microseal ‘B’ is presented in this paper as a cover seal for open-form microfluidic devices. This film is adaptable to high working temperatures and is biocompatible. The quality of the sealing film was investigated by leak tests, fluorescence tests, and contact angle measurements. The investigations revealed its sealing strength, fluorescence detection compatibility, and surface wettability. It was found that the proposed sealing polyester film on the 3D-printed device could sustain a gauge pressure of 2.7 atm at a flow rate of 4 mL/min without any leaks. It also provided fluorescence detection compatibility and an intensity-to-background ratio in the range of 2.3 to 4.5 for particle sizes of 5 μm and 15 μm, respectively, which is comparable with the performances of other sealing materials. The film’s hydrophobicity is comparable to other polymers used in microfluidics. This paper concludes by showcasing some applications of such transparent tops in classical microfluidic devices used for droplet generation and fluid mixing, in order to demonstrate the prospects of this fabrication technique in lab-on-a-chip devices.

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Section: Resultsmentioning

confidence: 99%

Demonstration of a Transparent and Adhesive Sealing Top for Microfluidic Lab-Chip Applications

Agarwal,

Salahuddin,

Ahamed

2024

Sensors

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“…Through the plasma oxygen system (deposition of chemical vapors enhanced with plasma), the device and the glass base are exposed for 3 min at a pressure of more than 4000 g overnight [ 18 , 19 , 20 ]. In the case of a droplet storage chip, multilevel photopolymer technology and microdevice manufacturing technology were used, as described in previous work [ 25 , 26 , 27 ].…”

Section: Methodsmentioning

confidence: 99%

Droplets for Gene Editing Using CRISPR-Cas9 and Clonal Selection Improvement Using Hydrogels

Pérez-Sosa,

Pérez,

Vallejo-Janeta

et al. 2024

Micromachines

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Gene editing tools have triggered a revolutionary transformation in the realms of cellular and molecular physiology, serving as a fundamental cornerstone for the evolution of disease models and assays in cell culture reactions, marked by various enhancements. Concurrently, microfluidics has emerged over recent decades as a versatile technology capable of elevating performance and reducing costs in daily experiments across diverse scientific disciplines, with a pronounced impact on cell biology. The amalgamation of these groundbreaking techniques holds the potential to amplify the generation of stable cell lines and the production of extracellular matrix hydrogels. These hydrogels, assuming a pivotal role in isolating cells at the single-cell level, facilitate a myriad of analyses. This study presents a novel method that seamlessly integrates CRISPR-Cas9 gene editing techniques with single-cell isolation methods in induced pluripotent stem cell (hiPSC) lines, utilizing the combined power of droplets and hydrogels. This innovative approach is designed to optimize clonal selection, thereby concurrently reducing costs and the time required for generating a stable genetically modified cell line. By bridging the advancements in gene editing and microfluidic technologies, our approach not only holds significant promise for the development of disease models and assays but also addresses the crucial need for efficient single-cell isolation. This integration contributes to streamlining processes, making it a transformative method with implications for enhancing the efficiency and cost-effectiveness of stable cell line generation. As we navigate the intersection of gene editing and microfluidics, our study marks a significant stride toward innovative methodologies in the dynamic landscape of cellular and molecular physiology research.

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“…Three main steps comprise the PDMS microfluidic device manufacturing process: (1) Photopolymer flexographic master mold (Fmold) fabrication, (2) manufacture of a reusable epoxy resin male mold (ERmold) from the FMold by a copy of the mold, and (3) PDMS replica from ERmold features ( Figure 1 ). The complete methodology was developed by the group and has been described in previous works [ 29 , 30 , 31 ].…”

Section: Methodsmentioning

confidence: 99%

Large Area Microfluidic Bioreactor for Production of Recombinant Protein

et al. 2022

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To produce innovative biopharmaceuticals, highly flexible, adaptable, robust, and affordable bioprocess platforms for bioreactors are essential. In this article, we describe the development of a large-area microfluidic bioreactor (LM bioreactor) for mammalian cell culture that works at laminar flow and perfusion conditions. The 184 cm2 32 cisterns LM bioreactor is the largest polydimethylsiloxane (PDMS) microfluidic device fabricated by photopolymer flexographic master mold methodology, reaching a final volume of 2.8 mL. The LM bioreactor was connected to a syringe pump system for culture media perfusion, and the cells’ culture was monitored by photomicrograph imaging. CHO-ahIFN-α2b adherent cell line expressing the anti-hIFN-a2b recombinant scFv-Fc monoclonal antibody (mAb) for the treatment of systemic lupus erythematosus were cultured on the LM bioreactor. Cell culture and mAb production in the LM bioreactor could be sustained for 18 days. Moreover, the anti-hIFN-a2b produced in the LM bioreactor showed higher affinity and neutralizing antiproliferative activity compared to those mAbs produced in the control condition. We demonstrate for the first-time, a large area microfluidic bioreactor for mammalian cell culture that enables a controlled microenvironment suitable for the development of high-quality biologics with potential for therapeutic use.

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Novel Reproducible Manufacturing and Reversible Sealing Method for Microfluidic Devices

Cited by 8 publications

References 18 publications

Demonstration of a Transparent and Adhesive Sealing Top for Microfluidic Lab-Chip Applications

Demonstration of a Transparent and Adhesive Sealing Top for Microfluidic Lab-Chip Applications

Droplets for Gene Editing Using CRISPR-Cas9 and Clonal Selection Improvement Using Hydrogels

Large Area Microfluidic Bioreactor for Production of Recombinant Protein

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