Review
Biochemical Research Methods
Nan Lu, Hui Min Tay, Chayakorn Petchakup, Linwei He, Lingyan Gong, Kay Khine Maw, Sheng Yuan Leong, Wan Wei Lok, Hong Boon Ong, Ruya Guo, King Ho Holden Li, Han Wei Hou
Summary: Blood tests are standard clinical procedures for disease and health condition screening, but the complex cellular background and biomolecular composition often make accurate analysis challenging. Label-free microfluidic technologies that rely on intrinsic cell properties are emerging as a promising approach for point-of-care blood diagnostics. This review summarizes recent advances in label-free blood cell separation, single cell analysis, and the combination of microfluidics with machine learning for enhanced clinical studies.
Article
Chemistry, Physical
Fangteng Song, Chao Wang, Chunhua Wang, Jianbo Wang, Yu Wu, Yihe Wang, Hong Liu, Yu Zhang, Lin Han
Summary: This study developed an integrated microfluidic platform for the multiplexed outline of exosome secretion by single cells, exploring intercellular heterogeneity and identifying unique secretion patterns in different cell subsets.
Article
Biotechnology & Applied Microbiology
Felix J. Hartmann, Dunja Mrdjen, Erin McCaffrey, David R. Glass, Noah F. Greenwald, Anusha Bharadwaj, Zumana Khair, Sanne G. S. Verberk, Alex Baranski, Reema Baskar, William Graf, David Van Valen, Jan Van den Bossche, Michael Angelo, Sean C. Bendall
Summary: This antibody-based method allows profiling of metabolic protein expression and regulation in single cells using mass spectrometry while characterizing cellular phenotype. The approach was validated using mass cytometry and applied to clinical samples, revealing tissue-restricted, metabolically repressed immune cells in human colorectal carcinoma. The combination with multiplexed ion beam imaging unveiled the spatial organization of metabolic programs in human tissues.
NATURE BIOTECHNOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Linmei Li, Haoran Su, Yahui Ji, Fengjiao Zhu, Jiu Deng, Xue Bai, Huibing Li, Xianming Liu, Yong Luo, Bingcheng Lin, Tingjiao Liu, Yao Lu
Summary: Cell-cell interactions are essential for regulating cellular activities. This study presents an integrative single-cell secretion analysis platform that profiles different secreted factors, spatial distances, and migration information simultaneously at the single-cell level. The analysis of tumor-stromal and tumor-immune interactions reveals that the initial distances between cells determine their migratory distances and direction. Close proximity enhances protein secretions while attenuating extracellular vesicle (EV) secretions. Migration has a stronger correlation with protein secretions than EV secretions, with absolute migration distance significantly affecting protein secretions. These findings emphasize the importance of spatial organization in regulating cell signaling behaviors and demonstrate the utility of the platform for comprehensive understanding of intercellular communication and interactions in the tumor microenvironment.
Article
Chemistry, Analytical
Lili Cong, Yu Tian, Zepeng Huo, Weiqing Xu, Chunxi Hou, Wei Shi, Weigang Wang, Chongyang Liang, Shuping Xu
Summary: This study introduces a microfluidic droplet-based fluorescence imaging platform for analyzing single cell-secreted VEGF, exploring cellular physiological clues at the single-cell level. By developing silica nanoparticle-based immunoprobes and conducting immunosandwich assays, the method demonstrates cellular heterogeneity in cytokine secretion.
ANALYTICAL CHEMISTRY
(2022)
Article
Multidisciplinary Sciences
Zhigang Chen, Yi Zhang, Wenlu Li, Chenyi Gao, Fengbo Huang, Lu Cheng, Menglei Jin, Xiaoming Xu, Jian Huang
Summary: This study utilizes single cell RNA-seq to investigate the mechanisms of fibroadenoma (FA) and develops organoid models for studying treatment resistance. The study identifies dysregulation of epithelial cells in FAs and discovers hormone-responsive functional signatures as well as mechanisms of hormone resistance. The authors also develop a human expandable FA organoid system and find that combinations of inhibitors can suppress the viability of tamoxifen-resistant organoids.
NATURE COMMUNICATIONS
(2023)
Article
Mathematical & Computational Biology
Junjing Cai, Qiwei Wang, Ce Wang, Yu Deng
Summary: Single cell dispensing techniques include limiting dilution, FACS, and microfluidic approaches. This paper presents a nearly non-destructive single-cell dispensing method based on object detection algorithm. The proposed method utilizes an automated image acquisition system and the PP-YOLO neural network model for detection. Experimental results show that the model achieves a good balance of detection speed and accuracy, inferring a 320 x 320 pixel image in at least 0.9 ms with a precision of 98.6% on a NVidia A100 GPU.
MATHEMATICAL BIOSCIENCES AND ENGINEERING
(2023)
Article
Cell Biology
Jie Yang, Yongyun Li, Yanping Han, Yiyi Feng, Min Zhou, Chunyan Zong, Xiaoyu He, Renbing Jia, Xiaofang Xu, Jiayan Fan
Summary: This study utilized single-cell RNA sequencing to uncover the cellular heterogeneity and developmental mechanism of human retinoblastoma, identifying two major cell types and 5 different cell states. Cone precursors were found to be the initiating cells of retinoblastoma, while UBE2C was associated with an activation state in the malignant progression of the tumor.
CELL DEATH & DISEASE
(2021)
Review
Biochemical Research Methods
Carlos Honrado, Paolo Bisegna, Nathan S. Swami, Federica Caselli
Summary: Microfluidic impedance cytometry is a label-free and high-throughput method for stratifying cellular systems based on electrophysiology, with applications ranging from life science research to precision medicine. Novel chip designs and data analytic strategies are laying the foundation for multiparametric cell characterization and subpopulation distinction, essential for understanding biological function and disease progression. Emerging developments in the technique, such as device design, data analytics, and phenotyping applications, offer exciting prospects for future challenges and directions in label-free quantification and isolation of subpopulations in heterogeneous biosystems.
Review
Chemistry, Multidisciplinary
Lu Huang, Yin Chen, Jianhua Zhou
Summary: Heterogeneous cell-cell communications are crucial for regulating biological processes and diseases. Traditional cell co-culture methods provide bulk-averaged results, while single-cell co-culture based on microfluidic platforms can reveal heterogeneous interactions between different cell types at single-cell resolution.
CELL REPORTS PHYSICAL SCIENCE
(2022)
Article
Chemistry, Analytical
Yingnan Sun, Qingqing Tian, Yongshu Liu, Kunming Xing, Yuyan Li, Yumin Liu, Shusheng Zhang
Summary: This study developed a microfluidic platform based on surface energy well (SEW) to achieve high spatiotemporal-resolved signal profiling at the single-cell level. The platform allows for controlled droplet replacement, reliable single-cell patterning, and successive treatment of adherent cells with reagent droplets.
Article
Multidisciplinary Sciences
Karen Davey, Daniel Wong, Filip Konopacki, Eugene Kwa, Tony Ly, Heike Fiegler, Christopher R. Sibley
Summary: The Nadia Instrument is a flexible, cost-effective, and user-friendly droplet-based microfluidics system capable of capturing a large number of single cells or nuclei's 3' mRNA in a single run. It demonstrates highly reproducible droplet size, low doublet rates, and high mRNA capture efficiency. When combined with different buffers and barcoded bead configurations, it enables diverse applications and the identification of distinct cell populations.
SCIENTIFIC REPORTS
(2021)
Review
Biochemical Research Methods
Shichao Lin, Yilong Liu, Mingxia Zhang, Xing Xu, Yingwen Chen, Huimin Zhang, Chaoyong Yang
Summary: Cells are the fundamental units of life, exhibiting vast heterogeneity. Single-cell transcriptomics provides insights into cell-to-cell gene expression variabilities. Microfluidic technologies have significantly advanced single-cell transcriptomics, improving throughput and comprehensiveness. Additionally, single-cell transcriptomics is expanding into spatial and temporal analysis.
Article
Engineering, Biomedical
Chao Wang, Chunhua Wang, Yu Wu, Jianwei Gao, Yingkuan Han, Yujin Chu, Le Qiang, Jiaoyan Qiu, Yakun Gao, Yanhao Wang, Fangteng Song, Yihe Wang, Xiaowei Shao, Yu Zhang, Lin Han
Summary: This work presents a high-throughput platform for living single-cell multi-index secreted biomarker profiling, combined with machine learning for accurate tumor cell classification, offering new intelligent tools for cancer research and biomedical studies.
ADVANCED HEALTHCARE MATERIALS
(2022)
Article
Biochemical Research Methods
Elizabeth E. Crouch, Loukas N. Diafos, Edward J. Valenzuela, Kaylee Wedderburn-Pugh, Janeth Ochoa Birrueta, Jaela Caston, Tara Joseph, Madeline G. Andrews, Aparna Bhaduri, Eric J. Huang
Summary: Angiogenesis and neurogenesis are functionally interconnected during brain development. This article presents a method to purify prenatal human brain vascular cells using FACS and utilize them in various downstream applications. The method is simple and efficient, generating high yields from small amounts of tissue. These vascular cells can be cultured and used in various in vitro assays. Additionally, the article describes how to use these cells for transplantation into cortical organoids to study neurovascular interactions in prenatal human brain development.
