SENSORS AND ACTUATORS B-CHEMICAL
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2020
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IF 7.1
Mostafa Gouda,Kai Chen,Xiaoli Li,Yufei Liu,Yong He
Microalgae single-cell electrochemical characteristics are playing an important key role in its industrial application and functionality. This study conducted a single-cell electrochemical study to explain the influence mechanism of intracellular phenolic and flavonoid contents of microalgae on its surface electric charge. In the method, four microalgae species, Arthrospira platensis (AP), Chlorella vulgaris (CV), Phaeodactylum tricornutum (PT), and Scenedesmus obliquus (SO), were studied through single-cell electrochemical current (SCC), single-cell micro-Raman spectrometry (SCMR), and other physicochemical methods. Also, they were correlated with the chemical analytical methods. In the results, PT and AP phenolics, flavonoids, and antioxidant activities were significantly high compared to CV and SO. On the other hand, SO had the highest SCC compared to AP and PT. Also, SCMR at 1514 cm −1 was negatively correlated with antioxidant activity by nitric oxide (r 2 = -0.80). In conclusion, single-cell current measurement by gold microelectrode can verify the antioxidant activity of the different microalgae species. Besides, this work provides a simple micro-level study of the single-cell phytochemicals to better explain their exact functional impacts compared to the methods of macro-analyses that use millions of cells, which reduces their sensitivity. In which, it can be applied in the real-time functionality evaluation during microalgae cultivation.
研究领域:
微生物学
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电化学
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分析化学
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光谱学
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生物技术
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Haimeng Pei,Haibin Si,Dexin Du,Lu Li,Bo Tang
Extracellular vesicles (EVs) play a crucial role in mediating cell–cell communication and shaping cellular functions. However, the relationship between EVs secretion and cellular drug sensitivity, as well as the role of EVs in drug resistance development, remains unclear. Therefore, characterizing rare drug-resistant cells within cell populations and obtaining time-resolved information on single-cell EVs secretion are essential for understanding EVs’ role in drug resistance. Herein, we utilize a single-cell EVs secretion quantification chip (SCQ-chip) to compartmentalize individual cells and longitudinally monitor their secretion during drug treatment. The results show that EVs secretion is closely related to cellular drug sensitivity and strongly depends on the drug dose. Low-dose drugs induce increased levels of EVs secretion as a protective mechanism, whereas high-dose drugs reduce both cell viability and EVs secretion. Notably, stem-like cells exhibit a unique slow-fluctuating EVs secretion pattern. Furthermore, coculturing drug-sensitive cells with EVs from drug-resistant cells revealed that EVs facilitate drug resistance transfer. Proteomic analysis indicates that this process is primarily driven by metabolism-related pathways, confirming EVs as mediators of drug resistance dissemination. Our results proposed that EVs secretion is a promising indicator for drug resistance analysis, which will inspire us to develop new drugs targeting EVs secretion as a strategy to overcome drug resistance.
研究领域:
分子生物学
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细胞生物学
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药理学
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生物化学
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癌症研究
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产品关联
DOI
PubMed
高影响因子
Cell Reports Physical Science
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2023
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IF 8.9
Lu Huang,Zhangcai Liu,Jinxu He,Juanhua Li,Zhihao Wang,Jianhua Zhou,Yin Chen
The assessment of cell migration and proliferation is essential in the field of oncology. It has been widely performed for cancer prognosis and the prediction of treatment outcome. The microdevice-based methods have enabled the assessment of these two processes at single-cell resolution, which could acquire unique information on cell heterogeneity and subtypes. However, most of the current platforms show limited throughput due to design defects or lack of modules for high-speed data analysis, which greatly hampers the extraction of statistically unbiased biological data. To address this challenge, we propose a high-throughput system consisting of an addressable dual-nested microwell array chip (DNMA chip) and an artificial intelligence (AI)-based image analysis algorithm. Our DNMA chip allows single-cell trapping, label-free encoding, and long-term incubation. Combined with AI-aided data processing, the migration and proliferation of single tumor cells under normal culture or chemotherapy are quantitatively analyzed in a high-throughput and non-destructive manner.
研究领域:
肿瘤学
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生物医学工程
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单细胞分析
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微流控技术
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转化医学
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人工智能在医学中的应用
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Binghan Li,Xiaoyue Shen,Rujia Liu,Yingjie Zhao,Hongli Cao,Dengchao Wang
Tryptophan levels in the human body are closely related to disease development and metabolic processes, but identification and quantification of tryptophan enantiomers at the single-cell level is still very challenging now. Herein, the mono-(6-ethanediamine-6-deoxy)-β-cyclodextrin (β-CD)-modified carbon nanopipet (CNP) was fabricated, and high-enantioselective electrochemical detection of tryptophan was achieved. Interestingly, the selectivity of the prepared CNP toward l-tryptophan (l-Trp) and d-Tryptophan (d-Trp) could be modulated by adjusting the solution pH. Moreover, besides measuring the l-Trp concentration in a preserved cell environment, the fabricated tip could also be used to monitor the dynamics of l-Trp metabolism in the presence of representative amino acids. This work would offer a new approach to measure Trp enantiomers and reveal tryptophan metabolic pathways at the single-cell level.
研究领域:
生物化学
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分析化学
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医学
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产品关联
DOI
PubMed
高影响因子
MOLECULES
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2023
·Vol. 28
·期 10
·4202
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IF 4.6
Pujia Zhu,Guowei Bu,Ruifeng Hu,Xianqin Ruan,Rongrong Fu,Zhourui Zhang,Qiongqiong Wan,Xin Liu,Yiliang Miao,Suming Chen
Mass spectrometry (MS)-based lipidomic has become a powerful tool for studying lipids in biological systems. However, lipidome analysis at the single-cell level remains a challenge. Here, we report a highly sensitive lipidomic workflow based on nanoflow liquid chromatography and trapped ion mobility spectrometry (TIMS)-MS. This approach enables the high-coverage identification of lipidome landscape at the single-oocyte level. By using the proposed method, comprehensive lipid changes in porcine oocytes during their maturation were revealed. The results provide valuable insights into the structural changes of lipid molecules during porcine oocyte maturation, highlighting the significance of sphingolipids and glycerophospholipids. This study offers a new approach to the single-cell lipidomic.Keywords:mass spectrometry;lipidomic;single-cell;oocytes;liquid chromatography
研究领域:
生殖生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Kang Liu,Shuohan Huang,Danjun Fang
Here, a microelectrode approach is established to measure the flip-flop rate of cholesterol in plasma membranes at single living cells. The initial validation is performed in a modeled phospholipid bilayer positioned at an interconnecting hole between two compartments, in which cholesterol in one compartment diffuses into the other one through a flip-flop movement in the bilayer and is then detected by a cholesterol oxidase-modified microelectrode. As compared with the time (140 ± 28 s) for free cholesterol transport in absence of the bilayer, a prolonged time (702 ± 42 s) is needed to observe the current increase in the presence of the bilayer. The difference in the time (562 s) gives the estimated flip-flop time of cholesterol in the bilayer. The position of the microelectrode in contact with a living cell and the injection of cholesterol inside the cell are further applied to measure the cholesterol flip-flop in the plasma membrane. The average time (1183 ± 146 s) is obtained to observe an additional current increase at the microelectrode, which reflects the cholesterol flip-flop rate in plasma membranes in single living cells. All these results support the establishment of this microelectrode approach for the study of the cholesterol flip-flop process in lipid membranes.
研究领域:
生物化学
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分析化学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2020
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IF 6.785
Zishuai Li,Zhengmao Wang,Junmin Pan,Xiaoxiao Ma,Wenpeng Zhang,Zheng Ouyang
Single-cell metabolite analysis plays an important role in biological study. While mass spectrometry is a powerful tool for identification and quantitation of metabolites, the low absolute analyte amounts in single cell and difficulty in sampling represent significant challenges in single cell analysis. In this study, we developed an effective method with a simple sampling procedure for analyzing single cells. A single cell was driven to a capillary tip through electro-migration, followed by releasing the cell contents through electroporation, into a sealed small volume (∼1.5 pL) to prevent dilution. Subsequent mass spectrometry analysis was performed directly with nanoelectrospray ionization. This method was applied for analyzing a variety of cells and monitoring the metabolic changes in response to perturbed cell culturing conditions. This method opens a new avenue for easy and rapid analysis of single cells with high sensitivity.
研究领域:
生物化学
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分析化学
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细胞生物学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2024
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IF 6.7
Wenxiao Tang,Beibei Chen,Man He,Gaofei Song,Yonghong Bi,Bin Hu
Understanding the interactions between mercury and microalgae, especially the interactions between inorganic mercury (IHg) and extracellular polymeric substances (EPS, a protective barrier between cells and their external environment), is essential for elucidating mercury’s toxicological mechanisms. Given the inherent cell heterogeneity, a novel analysis system of an online viscoelastic fluid focusing chip-time-resolved analysis inductively coupled plasma mass spectrometry has been developed to investigate the bioaccumulation of HgS nanoparticles and Hg2+ in single Microcystis aeruginosa (M. aeruginosa) cells, exploring the interaction mechanisms between HgS/Hg2+ accumulation in algal cells and EPS. The single-cell analysis results reveal minimal bioavailability of HgS within algal cells, with mercury’s toxicity to M. aeruginosa being species-dependent. Notably, algal cells exhibited more heterogeneity in HgS uptake than in Hg2+ uptake. Under Hg2+/HgS stress, M. aeruginosa cells with EPS removed (EPS-R algal cells) showed an increased level of bioaccumulation of mercury compared to those with EPS (EPS-C algal cells), highlighting the critical role of EPS in mercury bioaccumulation. Overall, the designed viscoelastic fluid microfluidic focusing chip integrates focusing and cleaning functions, featuring easy fabrication, simple operation, low sample loss, and relatively high throughput. Under the optimal conditions, the sample throughput is 1195 min–1 and the cell recovery is 90%. Besides, this research offers novel insights into the interaction mechanisms between Hg2+/HgS and EPS in microalgal cells and unveils the specific toxic effects of Hg2+/HgS on M. aeruginosa at the single-cell level, contributing to a deeper understanding of mercury’s ecological and toxicological impact in aquatic environments.
研究领域:
环境科学
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毒理学
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分析化学
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产品关联
DOI
PubMed
高影响因子
Science China-Chemistry
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2024
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IF 10.4
Xu Xing,Zeng Xi,Lin Xin,Lin Shiyan,Liang Shanshan,Tian Tian,Su Rui,Song Jia,Yang Chaoyong
Single-cell joint analysis of methylome and transcriptome reveals how the methylation regulates the transcriptional activity. However, traditional bench-top protocols for single-cell DNA methylation and RNA transcription co-detection are labor-intensive, cost-ineffective and contaminant-prone. Herein, we establish the DMF-scMT-seq, a highly-efficient and cost-effective method to simultaneously analyze single-cell DNA methylation and transcriptional activity based on digital microfluidics. DMF-scMT-seq automates the workflow of single-cell isolation, cellular hypotonic lysis, nucleic acid separation and methylome/transcriptome library construction in a contactless and addressable way. The system ensures high accuracy ( R >0.85), high gene detection ability (14,697 genes per cell at 4 million sequencing depth), and high CpG coverage (677,198 CpG sites per cell at 1 million sequencing depth). By using DMF-scMT-seq, the relationship of DNA methylation and RNA transcription under different genomic contexts is resolved. We further apply DMF-scMT-seq to study the dynamics of transcription regulation with methylation-inhibiting anti-tumor Decitabine, and identify the methylated promoter/gene body driven genes in response to Decitabine treatment. DMF-scMT-seq facilitates the construction of the correlation of DNA methylation and transcriptional activity at the single-cell level in a flexible, sensitive and accurate way, which is anticipated to be a powerful tool in studying single-cell biological systems.
研究领域:
生物信息学
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表观遗传学
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分子生物学
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DOI
高影响因子
Clinical and Translational Medicine
·
2023
·Vol. 13
·期 6
·e1297
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IF 10.6
Jie Yang,Hu Tan,Mengjia Sun,Renzheng Chen,Zhao Jian,Yuanbin Song,Jihang Zhang,Shizhu Bian,Bo Zhang,Yi Zhang,Xubin Gao,Zhen Chen,Boji Wu,Xiaowei Ye,Hailin Lv,Zhen Liu,Lan Huang
Background Atrial fibrillation (AF) is associated with an increased risk of thrombosis of the left atrial appendage (LAA). However, the molecular mechanisms underlying this site-specificity remain poorly understood. Here, we present a comparative single-cell transcriptional profile of paired atrial appendages from patients with AF and illustrate the chamber-specific properties of the main cell types. Methods Single-cell RNA sequencing analysis of matched atrial appendage samples from three patients with persistent AF was evaluated by 10× genomics. The AF mice model was created using Tbx5 knockout mice. Validation experiments were performed by glutathione S-transferase pull-down assays, coimmunoprecipitation (Co-IP), cleavage assays and shear stress experiments in vitro. Results In LAA, phenotype switching from endothelial cells to fibroblasts and inflammation associated with proinflammatory macrophage infiltration were observed. Importantly, the coagulation cascade is highly enriched in LAA endocardial endothelial cells (EECs), accompanying the up-regulation of a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1) and the down-regulation of the tissue factor pathway inhibitor (TFPI) and TFPI2. Similar alterations were verified in an AF mouse model ( Tbx5 +/− ) and EECs treated with simulated AF shear stress in vitro. Furthermore, we revealed that the cleavage of both TFPI and TFPI2 based on their interaction with ADAMTS1 would lead to loss of anticoagulant activities of EECs. Conclusions This study highlights the decrease in the anticoagulant status of EECs in LAA as a potential mechanism underlying the propensity for thrombosis, which may aid the development of anticoagulation therapeutic approaches targeting functionally distinct cell subsets or molecules during AF.
研究领域:
心血管病学
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分子生物学
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基因组学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Huan Yao,Jinlei Yang,Zhengmao Wang,Xingyu Pan,Junmin Pan,Hongmei Li,Sichun Zhang
Microalgae metabolite analysis is fundamental for the rational design of metabolic engineering strategies for the biosynthesis of high-value products. Mass spectrometry (MS) has been utilized for single-cell microalgae analysis. However, limitations in the detection throughput and polarities of detectable substances make it difficult to realize high-throughput screening of high-performance microalgae. Herein, a plasma-assisted label-free mass cytometry, named as PACyESI-MS, was proposed combining the advantages of orthogonal hybrid ionization and high-throughput MS analysis, which realized rapid metabolite detection of single microalgae. The cell detection throughput of PACyESI-MS was up to 52 cells/min. Dozens of the critical primary and secondary metabolites within single microalgae were detected simultaneously, including pigments, lipids, and energy metabolites. Furthermore, metabolite changes of Chlamydomonas reinhardtii and Haematococcus pluvialis under nitrogen deficiency stress were studied. Discrimination of Chlamydomonas under different nutrient conditions was realized using single-cell metabolite profiles obtained by PACyESI-MS. The relationships between the accumulation of bioactive astaxanthin and changes in functional primary metabolites of Haematococcus were investigated. It was demonstrated that PACyESI-MS can detect the flexible change of metabolites in single microalgae cells under different nutritional conditions and during the synthesis of high-value products, which is expected to become an important tool for the design of metabolic engineering-based high-performance microalgae factories.
研究领域:
生物化学
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代谢工程
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植物学
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产品关联
DOI
PubMed
高影响因子
Materials Today Bio
·
2025
·
IF 10.2
Haitao Liu,Tingting Tao,Zhongqiao Gan,Yingying Xie,Yaqing Wang,Yizhao Yang,Xu Zhang,Xianliang Li,Jianhua Qin
Cancer organoids have improved our understanding of recapitulating the histology, genotypes, and drug response of patient tumors for personalized medicine. However, the existing cancer organoids are typically grown in animal-derived matrices (e.g., Matrigel), which suffer from poor reproducibility and low throughput due to uncontrollable origin of seed cells, undefined matrix, and manual manipulation. Here, we report a new strategy to massively generate uniform pancreatic cancer organoids (PCOs) in a droplet system from single cells. This system is composed of all-in-water fluids that allow to mildly encapsulate single tumor cell into isolated droplet, which subsequently proliferate and self-assemble into organoids, resembling the initial state of a tumor in the body. This high-throughput method can produce thousands of organoids in a single batch. The droplets can serve as templates for synthesizing defined microgels with proper stiffness similar to that of native tumors, facilitating functional expressions of PCOs. These organoids exhibit superior uniformity and controllability in terms of size and morphologies compared with organoids cultured in manually dropped Matrigel, due mainly to the controllable number of initiating cells and defined microgels. In addition, the established organoids maintain the key biomarkers of pancreatic tumor (e.g., KRT7, KRT19 and SOX9) and higher expression of genes associated with drug metabolism confirmed by RNA-seq and PCR analysis. Furthermore, they show distinguishing responses to four clinically used drugs in a reproducible manner in automatic pipetting workstation, indicating the feasibility of the proposed method in high-throughput drug testing. The established strategy has integrated the formation, 3D cultures, and analysis of PCOs derived from single cells in a whole system, which may provide a novel platform for advancing organoids research with standardized procedure in translational applications.
研究领域:
肿瘤学
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再生医学
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生物工程
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）