The use of droplet-based microfluidic technologies for accelerated selection of Yarrowia lipolytica and Phaffia rhodozyma yeast mutants DOI Creative Commons
Taras Mika, Mārtiņš Kalniņš, Krišs Spalviņš

et al.

Biology Methods and Protocols, Journal Year: 2024, Volume and Issue: 9(1)

Published: Jan. 1, 2024

Abstract Microorganisms are widely used for the industrial production of various valuable products, such as pharmaceuticals, food and beverages, biofuels, enzymes, amino acids, vaccines, etc. Research is constantly carried out to improve their properties, mainly increase productivity efficiency reduce cost processes. The selection microorganisms with improved qualities takes a lot time resources (both human material); therefore, this process itself needs optimization. In last two decades, microfluidics technology appeared in bioengineering, which allows manipulating small particles (from tens microns nanometre scale) flow liquid microchannels. based on small-volume objects (microdroplets from nano femtolitres), manipulated using microchip. chip made an optically transparent inert medium material contains series channels size (<1 mm) certain geometry. Based physical chemical properties microparticles (like size, weight, optical density, dielectric constant, etc.), they separated microsensors. idea accelerated application microfluidic technologies separate mutants after mutagenesis. This article discusses possible practical implementation separation mutants, including yeasts like Yarrowia lipolytica Phaffia rhodozyma mutagenesis will be discussed.

Language: Английский

Unveiling the Potential of Single‐Cell Encapsulation in Biomedical Applications: Current Advances and Future Perspectives DOI Creative Commons
Manuel Pires‐Santos,

Sara Nadine,

João F. Mano

et al.

Small Science, Journal Year: 2024, Volume and Issue: 4(5)

Published: March 25, 2024

The encapsulation of single cells has emerged as a promising field in recent years, owing to its potential applications cell‐based therapeutics, bioprinting, vitro cell culture, high‐throughput screening, and diagnostics. Single‐cell units offer several advantages, including compatibility with standard imaging techniques, superior diffusion rates, lower material‐to‐cell volume ratios. They also serve effective carriers for targeted drug delivery, allowing precise administration therapeutics cell‐mediated quantities. Moreover, single‐cell exhibit improved circulation throughout the vasculature, reduced likelihood entrapment compared multicell strategies. However, production from random dispersion follows Poisson distribution, requiring separation empty ones. Various methods have been developed address this challenge; nevertheless, majority these strategies are either expensive or time‐consuming. This review provides an in‐depth analysis advantages limitations their applications, well comprehensive overview most used techniques sorting

Language: Английский

Citations

11
Microfluidic platform for omics analysis on single cells with diverse morphology and size: A review DOI
Shujing Lin, Dan Feng, Xiao Han

et al.

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1294, P. 342217 - 342217

Published: Jan. 6, 2024

Language: Английский

Citations

7
Yeast-based screening platforms to understand and improve human health DOI
Marcus Deichmann, Frederik G. Hansson, Emil D. Jensen

et al.

Trends in biotechnology, Journal Year: 2024, Volume and Issue: 42(10), P. 1258 - 1272

Published: April 26, 2024

Language: Английский

Citations

7
Acoustofluidic separation of cell-encapsulated droplets based on traveling surface acoustic wave-induced acoustic radiation force DOI
Mushtaq Ali,

Song Ha Lee,

Beomseok Cha

et al.

Sensors and Actuators B Chemical, Journal Year: 2024, Volume and Issue: 415, P. 135988 - 135988

Published: May 17, 2024

Language: Английский

Citations

5
Label-free single-cell analysis in microdroplets using a light-scattering-based optofluidic chip DOI
Li Liang, Minhui Liang, Zewen Zuo

et al.

Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: 253, P. 116148 - 116148

Published: Feb. 20, 2024

Language: Английский

Citations

4
Droplet-based microfluidic platform for detecting agonistic peptides that are self-secreted by yeast expressing a G-protein-coupled receptor DOI Creative Commons

Ririka Asama,

Cher J. S. Liu,

Masahiro Tominaga

et al.

Microbial Cell Factories, Journal Year: 2024, Volume and Issue: 23(1)

Published: April 9, 2024

Abstract Background Single-cell droplet microfluidics is an important platform for high-throughput analyses and screening because it provides independent compartmentalized microenvironment reaction or cultivation by coencapsulating individual cells with various molecules in monodisperse microdroplets. In combination microbial biosensors, this technology becomes a potent tool the of mutant strains. study, we demonstrated that genetically engineered yeast strain can fluorescently sense agonist ligands via heterologous expression human G-protein-coupled receptor (GPCR) concurrently secrete candidate peptides highly compatible single-cell microfluidic new agonistically active peptides. Results The water-in-oil microdroplets were generated using flow-focusing chip to encapsulate coexpressing GPCR [i.e., angiotensin II type 1 (AGTR1)] secretory agonistic peptide (Ang II)]. single cultured droplets then observed under microscope analyzed image processing incorporating machine learning techniques. AGTR1-mediated signal transduction elicited self-secreted Ang was successfully detected fluorescent reporter cultures. system could also distinguish analog different activities. Notably, further culture enabled detection rarely existing positive II-secreting) model mixed cell library, whereas conventional batch-culture environment shake flask failed do so. Thus, our approach provided microculture environments, which prevent diffusion, dilution, cross-contamination secreted from easy identification agonists. Conclusions We established droplet-based integrated biosensor expressed This offers individually isolated microenvironments allow secreting these gaging their signaling activities, Our base on biosensors will be widely applicable metabolic engineering, environmental drug discovery.

Language: Английский

Citations

4
Recent advancements in single-cell metabolic analysis for pharmacological research DOI Creative Commons
Ying Hou,

Hongren Yao,

Jin‐Ming Lin

et al.

Journal of Pharmaceutical Analysis, Journal Year: 2023, Volume and Issue: 13(10), P. 1102 - 1116

Published: Aug. 23, 2023

Cellular heterogeneity is crucial for understanding tissue biology and disease pathophysiology. Pharmacological research being advanced by single-cell metabolic analysis, which offers a technique to identify variations in RNA, proteins, metabolites, drug molecules cells. In this review, the recent advancement of analysis techniques their applications metabolism response are summarized. High-precision controlled isolation manipulation provided microfluidics-based methods, such as droplet microfluidics, microchamber, open microfluidic probe, digital microfluidics. They used tandem with variety detection techniques, including optical imaging, Raman spectroscopy, electrochemical detection, RNA sequencing, mass spectrometry, evaluate changes administration. The advantages disadvantages different discussed along challenges future directions analysis. These employed pharmaceutical studying resistance pathway, therapeutic targets discovery, vitro model evaluation.

Language: Английский

Citations

11
Single‐Cell Liquid‐Core Microcapsules for Biomedical Applications DOI
Manuel Pires‐Santos, Mariana Carreira, Bruno P. Morais

et al.

Advanced Healthcare Materials, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 24, 2025

Abstract More recently, single‐cell encapsulation emerged as a promising field in biomedicine due to its potential applications, cell analysis and therapy. Traditional techniques involve embedding cells crosslinked polymers create continuous microgels, suitable mainly for adherent cells, or encapsulating them droplets only short‐term analysis, their instability. In this study, we developed method single liquid‐core microcapsules address these limitations. The liquid system is generated an all aqueous environment through polymeric electrostatic interactions. Additionally, design innovative low cost sorting utilizing magnetic nanoparticles (MNPs) efficiently select encapsulated units further applications. This tested with both suspension types, demonstrating cytocompatibility no abnormal effects on behavior. MNP‐based achieved nearly 80% purity of the population. Overall, technology provides highly efficient such screening, by enabling precise short medium‐term real time monitoring, high resolution imaging cellular Furthermore, semipermeable membrane unlocks new advancing therapy offering protection while ensuring diffusion therapeutic factors, paving way strategies.

Language: Английский

Citations

0
Gray‐Level Guided Image‐Activated Droplet Sorter for Label‐Free, High‐Accuracy Screening of Single‐Cell on Demand DOI

Zhen Liu,

Yidi Zhang, Jianing Li

et al.

Small, Journal Year: 2025, Volume and Issue: unknown

Published: May 8, 2025

Abstract Single‐cell encapsulation in droplet microfluidics has become a powerful tool precision medicine, single‐cell analysis, and immunotherapy. However, generation with is random process, which also results large number of empty multi‐cell droplets. Current sorting technologies suffer from drawbacks such as fluorescent labeling, inability to remove droplets, or low throughput. This paper presents gray‐level guided image‐activated sorter (GL‐IADS), enables label‐free, high‐accuracy screening droplets by rejecting The based recognition method can accurately classify images (empty, single‐cell, droplets), especially differentiating cell‐laden (accuracy 100%). Crucially, this reduces the image processing time ≈300 µs, makes GL‐IADS possible reach an ultra‐high throughput up hundreds even KHz. integrates novel detachable acoustofluidic system, achieving purity 97.9%, 97.4%, >99% for multi‐cell, respectively, 43 Hz. holds promise numerous biological applications that are previously difficult fluorescence‐based technologies.

Language: Английский

Citations

0
Integration of acoustic, optical, and electrical methods in picoliter droplet microfluidics for rare particles enrichment DOI Creative Commons
Huasheng Zhuo, Chunhua He,

Canfeng Yang

et al.

Communications Engineering, Journal Year: 2025, Volume and Issue: 4(1)

Published: May 13, 2025

Rare particle enrichment plays a pivotal role in advancing numerous scientific research areas and industrial processes. Traditional methods encounter obstacles such as low efficiency, high cost, complexity. Acoustic focusing, optical fiber detection, electrical manipulation have shown potential microfluidics for analysis. This study pioneers the integration of acoustic, optical, units to overcome traditional limitations. The cooperative dynamics acoustic flow focusing are explored. fibers with an enhanced detection algorithm greatly boost sensitivity. Furthermore, droplet charging enhance tip phenomenon is complemented validated. sorting accuracy enriching large-size H22 cells reached 99.8% 99.3%, respectively, target cell concentration increased by nearly 86-fold. Our work significantly enhances sensitivity accuracy, ultimately offering robust reliable solution generating droplets enrich rare particles.

Language: Английский

Citations

0