Review
Chemistry, Analytical
Zhihang Yu, Jing Jin, Lingling Shui, Huaying Chen, Yonggang Zhu
Summary: Proteins play a crucial role in cellular activities, and single-cell protein research is vital for understanding cellular heterogeneity. Droplet technology, as an effective tool for single-cell manipulation, has revolutionized high-throughput single-cell protein analysis by precisely controlling droplet volume and preventing contamination. The high-throughput and automated nature of droplet technology allows for the analysis of tens of thousands of single cells within an hour, making it highly compatible and admired in the field of single-cell protein analysis.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2021)
Review
Chemistry, Analytical
Zhenqi Jiang, Haoran Shi, Xiaoying Tang, Jieling Qin
Summary: Single-cell manipulation and analysis is crucial for studying biological processes and cellular heterogeneity. Droplet microfluidics has revolutionized single-cell studies by allowing extraction of information at the genomic, transcriptomic, proteomic, and metabolomic level from individual cells. It provides a detailed understanding of intracellular heterogeneity and changes in cellular phenotype. This review summarizes the advances of droplet microfluidics in single-cell analysis, emphasizing its advantages, preparation, and various assays. It also highlights its application in studying gene expression and other biomolecules at the single-cell level, facilitating fundamental and clinical research.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2023)
Article
Biochemical Research Methods
Alina Peethan, M. Aravind, Santhosh Chidangil, Sajan D. George
Summary: Droplet splitting using tailored surface wettability is an important technique for creating ultralow volumes of samples, which have various applications in bioassays, tissue engineering, and material synthesis. In this work, a non-adhesive contrast quartz substrate is fabricated for the trapping and analysis of single biological cells using Raman spectroscopy. The substrate allows the splitting of microliter droplets into smaller daughter droplets, enabling high throughput screening and analysis of single cells.
Article
Chemistry, Multidisciplinary
Chunyu Li, Yanhai Gong, Xixian Wang, Jian Xu, Bo Ma
Summary: An addressable dynamic droplet array (aDDA) integrates the advantages of static droplet arrays and continuous-flow droplet platforms, allowing all sample preparation steps to be performed within a sub-nanoliter droplet. Validation studies show that aDDA reduces amplification bias and increases genome recovery efficiency, making it a valuable tool for high-genome-coverage sequencing of single microbial cells.
Article
Cell Biology
Mahsan Banijamali, Pontus Hojer, Abel Nagy, Petra Haag, Elizabeth Paz Gomero, Christiane Stiller, Vitaliy O. Kaminskyy, Simon Ekman, Rolf Lewensohn, Amelie Eriksson Karlstrom, Kristina Viktorsson, Afshin Ahmadian
Summary: Small extracellular vesicles (sEVs) have emerged as a valuable source of biomarkers for disease diagnosis and therapy monitoring. A new method called droplet barcode sequencing for protein analysis (DBS-Pro) has been developed to profile surface proteins on individual sEVs, enabling the identification of sEV subtypes. This method uses DNA barcoded affinity reagents and sequencing as readout to analyze multiple proteins in high throughput single vesicle profiling.
JOURNAL OF EXTRACELLULAR VESICLES
(2022)
Article
Chemistry, Multidisciplinary
Reem Khojah, Zhuyun Xiao, Mohanchandra K. Panduranga, Michael Bogumil, Yilian Wang, Maite Goiriena-Goikoetxea, Rajesh V. Chopdekar, Jeffrey Bokor, Gregory P. Carman, Rob N. Candler, Dino Di Carlo
Summary: Programming magnetic fields at the microscale using magnetostrictive materials coupled with ferroelectric materials enables refined control of magnetization, allowing for the manipulation of magnetic beads and cells with sub-micrometer precision.
ADVANCED MATERIALS
(2021)
Article
Genetics & Heredity
David Conchouso, Amani Al-Ma'abadi, Hayedeh Behzad, Mohammed Alarawi, Masahito Hosokawa, Yohei Nishikawa, Haruko Takeyama, Katsuhiko Mineta, Takashi Gojobori
Summary: In this study, three droplet-based microfluidic technologies were integrated into the metagenomic pipeline to enable high-throughput functional screening of bioproducts. By encapsulating single cells, integrating fluorescence-exciting lasers, and incorporating a passive droplet merger, researchers can quickly utilize microfluidic technologies for single-cell resolution and high-throughput capability (> 50,000 cells/day) in mining and bioprospecting metagenomic data.
GENOMICS PROTEOMICS & BIOINFORMATICS
(2021)
Article
Nanoscience & Nanotechnology
Akihiro Isozaki, Dunhou Huang, Yuta Nakagawa, Keisuke Goda
Summary: Droplet microfluidics is a powerful tool for various applications, with on-chip multiplexed droplet sorting allowing real-time and high-throughput sorting of cells with different phenotypes. The use of sequentially addressable dielectrophoretic arrays enables efficient and scalable droplet sorting.
MICROFLUIDICS AND NANOFLUIDICS
(2021)
Article
Chemistry, Multidisciplinary
Shinya Yamahira, Ryuji Misawa, Takahiro Kosaka, Mondong Tan, Shin Izuta, Hayato Yamashita, Yuji Heike, Akimitsu Okamoto, Teruyuki Nagamune, Satoshi Yamaguchi
Summary: This study presents a method for accurately positioning multiple types of cells using a photoactivatable material. The material can anchor cells through lipid-cell membrane interactions, and the anchoring can be activated using light exposure. The technique holds promise for high-throughput analysis of individual cell-cell communication.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2022)
Article
Chemistry, Multidisciplinary
Kun Yin, Shanqing Huang, Weidong Ruan, Haopai Wei, He Dong, Dianyi Liang, Meijuan Zhao, Wei Qi, Chaoyong Yang, Zhi Zhu
Summary: Iso-seq is a rapid and uniform single-cell whole-genome sequencing method that uses a specially designed dual-well structure and a capillary-based centrifugal-driven droplet generator to isolate single cells and compartmentalize their genomes in droplets within 3 minutes. Iso-seq achieves high mapping ratio, broad coverage of the genome, and low coefficient of variation, making it promising for a wide variety of biological applications.
Article
Chemistry, Multidisciplinary
Serban Dobroiu, Falco C. M. J. M. van Delft, Ayyappasamy Sudalaiyadum Perumal, Shantoshini Dash, Jenny Aveyard, Jeroen van Zijl, Jaap Snijder, Eric van den Heuvel, Jurgen van Berkum, Marie Pierre Blanchard, Cyril Favard, Dan Nicolau
Summary: One challenge in biosensing technology is detecting very low concentrations of analytes. The fluorescence interference contrast (FLIC) technique improves sensitivity by selectively amplifying or suppressing the emission of a fluorophore-labeled biomolecule. FLIC's extreme sensitivity to wavelength can be a problem due to variations in the fluorophore's vertical position.
Article
Chemistry, Multidisciplinary
Tianze Xie, Qiang Zhang, Weifei Zhang, Shuo Feng, Jin-Ming Lin
Summary: Cell adhesion is crucial for cellular behaviors and life processes, but studying specific cells is challenging due to adhesion deficiencies or interference from surrounding cells. This study presents an open microfluidic system for culturing adherent single cells, offering high versatility and compatibility for various assaying methods.
Review
Chemistry, Multidisciplinary
Haiyang Xie, Xianting Ding
Summary: The advancement of single-cell protein analysis technologies enables high-throughput and multiplexed protein measurements, providing a more comprehensive exploration of cellular signaling processes in combination with RNA sequencing and mass spectrometry. While the potential applications are vast, there are still limitations and challenges to be addressed in this field.
Article
Chemistry, Multidisciplinary
Haijun Cui, Xianxian Wang, Janine Wesslowski, Tina Tronser, Jakob Rosenbauer, Alexander Schug, Gary Davidson, Anna A. Popova, Pavel A. Levkin
Summary: Artificial multicellular systems are important in tissue engineering and regenerative medicine. The droplet-fusion technique developed in this study allows for high-throughput assembly of multiple cell spheroids into complex multicellular architectures, which can be used to study various biological processes such as cell signaling, cancer invasion, embryogenesis, and neural development.
ADVANCED MATERIALS
(2021)
Review
Biotechnology & Applied Microbiology
B. M. Tiemeijer, J. Tel
Summary: Single-cell techniques, especially microfluidics and microgel technology, are playing an increasingly important role in cell biological research. This review provides a comprehensive overview of how different types of hydrogels can be incorporated into droplet-based methods, offering novel and reliable analytic and screening applications in cell biology research.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Analytical
Ali Salehi-Reyhani, Frank Gesellchen, Dileep Mampallil, Rab Wilson, Julien Reboud, Oscar Ces, Keith R. Willison, Jonathan M. Cooper, David R. Klug
ANALYTICAL CHEMISTRY
(2015)
Article
Multidisciplinary Sciences
Eszter Lakatos, Ali Salehi-Reyhani, Michael Barclay, Michael P. H. Stumpf, David R. Klug
Article
Chemistry, Analytical
Agostino Cilibrizzi, Mattia Terenghi, Marina Fedorova, Rudiger Woscholski, David Klug, Ramon Vilar
SENSORS AND ACTUATORS B-CHEMICAL
(2017)
Meeting Abstract
Biophysics
Bogachan Tahirbegi, Alastair J. Magness, Aurelien Boillat, Keith R. Willison, David R. Klug, Thomas Knopfel, Liming Ying
BIOPHYSICAL JOURNAL
(2018)
Article
Chemistry, Physical
Hugh Sowley, ZhiQiang Liu, Julia Davies, Robert Peach, Rui Guo, Sophie Sim, FengQin Long, Geoffrey Holdgate, Keith Willison, Wei Zhuang, David R. Klug
JOURNAL OF PHYSICAL CHEMISTRY B
(2019)
Article
Multidisciplinary Sciences
Bogachan Tahirbegi, Alastair J. Magness, Maria Elena Piersimoni, Thomas Knopfel, Keith R. Willison, David R. Klug, Liming Ying
SCIENTIFIC REPORTS
(2020)
Article
Physics, Multidisciplinary
Taran Driver, Bridgette Cooper, Ruth Ayers, Ruediger Pipkorn, Serguei Patchkovskii, Vitali Averbukh, David R. Klug, Jon P. Marangos, Leszek J. Frasinski, Marina Edelson-Averbukh
Article
Cell Biology
Stelios Chrysostomou, Rajat Roy, Filippo Prischi, Lucksamon Thamlikitkul, Kathryn L. Chapman, Uwais Mufti, Robert Peach, Laifeng Ding, David Hancock, Christopher Moore, Miriam Molina-Arcas, Francesco Mauri, David J. Pinato, Joel M. Abrahams, Silvia Ottaviani, Leandro Castellano, Georgios Giamas, Jennifer Pascoe, Devmini Moonamale, Sarah Pirrie, Claire Gaunt, Lucinda Billingham, Neil M. Steven, Michael Cullen, David Hrouda, Mathias Winkler, John Post, Philip Cohen, Seth J. Salpeter, Vered Bar, Adi Zundelevich, Shay Golan, Dan Leibovici, Romain Lara, David R. Klug, Sophia N. Yaliraki, Mauricio Barahona, Yulan Wang, Julian Downward, J. Mark Skehel, Maruf M. U. Ali, Michael J. Seckl, Olivier E. Pardo
Summary: The study identified RSK4 as a driver of drug resistance and metastasis in lung and bladder cancer cells, and silencing this kinase increased sensitivity to chemotherapy and reduced metastasis. Drug screening revealed fluoroquinolone antibiotics as potent RSK4 inhibitors, with trovafloxacin replicating effects of RSK4 silencing. Further structural analysis provided insights into how trovafloxacin blocks RSK4 kinase activation. Additionally, patients receiving chemotherapy and prophylactic levofloxacin showed significantly prolonged overall survival, suggesting RSK4 inhibition as a potential therapeutic strategy for lung and bladder cancer.
SCIENCE TRANSLATIONAL MEDICINE
(2021)
Article
Chemistry, Analytical
Suhuur Osman, Claus Bendtsen, Samantha Peel, Linda Yrlid, Daniel Muthas, John Simpson, Keith R. Willison, David R. Klug
Summary: The cellular thermal shift assay (CETSA) is widely used to study drug-target engagement, especially in early stage drug discovery. Single-cell CETSA can accurately count active FOXO1 molecules and observe its stability and destabilization in the presence of small molecule inhibitors. The successful use of the MAC chip for single-cell CETSA enables the study of precious clinical samples and underlying population heterogeneity in cellular systems.
ANALYTICAL CHEMISTRY
(2021)
Article
Chemistry, Multidisciplinary
Bogachan Tahirbegi, Alastair J. Magness, Maria Elena Piersimoni, Xiangyu Teng, James Hooper, Yuan Guo, Thomas Knopfel, Keith R. Willison, David R. Klug, Liming Ying
Summary: Aggregation kinetics of proteins and peptides have been extensively studied for their importance in human diseases and physiological processes. However, most studies rely on bulk measurements and cannot directly measure assembly reactions at low protein concentrations. Using single-molecule methods and machine learning models, we developed a high-throughput protocol to study protein oligomers' stoichiometry and aggregation kinetics. Our results show promising classification accuracy.
FRONTIERS IN CHEMISTRY
(2022)
Meeting Abstract
Biochemistry & Molecular Biology
F. Prischi, S. Chrysostomou, R. Roy, K. Chapman, U. Mufti, R. Peach, L. Ding, F. Mauri, G. Bellezza, L. Cagini, M. Barbareschi, S. Ferrero, J. Abrahams, S. Ottaviani, L. Castellano, G. Giamas, J. Pascoe, D. Moonamale, L. Billingham, M. Cullen, D. Hrouda, M. Winkler, D. Klug, S. Yaliraki, M. Barahona, Y. Wang, M. Ali, M. Seckl, O. Pardo
Meeting Abstract
Oncology
Stelios Chrysostomou, Rajat Roy, Filippo Prischi, Katie Chapman, Uwais Mufti, Francesco Mauri, Guido Bellezza, Joel Abrahams, Silvia Ottaviani, Leandro Castellano, Georgios Giamas, David Hrouda, Mathias Winkler, David Klug, Sophia Yaliraki, Mauricio Barahona, Yulan Wang, Maruf Ali, Michael Seckl, Olivier Pardo
Meeting Abstract
Chemistry, Multidisciplinary
Sophie Sim, Hugh Sowley, Nathan Kidley, Laura Barter, David Klug
ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY
(2017)
Article
Oncology
Alastair J. Magness, James A. Squires, Beatrice Griffiths, Khurum Khan, Amanda Swain, Keith R. Willison, David Cunningham, Marco Gerlinger, David R. Klug
CONVERGENT SCIENCE PHYSICAL ONCOLOGY
(2017)
Meeting Abstract
Biophysics
K. R. Willison, A. Salehi-Reyhani, E. Burgin, M. Barclay, A. Brown, M. A. Neil, O. Ces, D. R. Klug
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
(2015)
