Discriminating Leukemia Cellular Heterogeneity and Screening Metabolite Biomarker Candidates using Label-Free Mass Cytometry

Author(s):  
Huan Yao ◽  
Hansen Zhao ◽  
Xingyu Pan ◽  
Xu Zhao ◽  
Jiaxin Feng ◽  
...  
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Soo-Yeon Cho ◽  
Xun Gong ◽  
Volodymyr B. Koman ◽  
Matthias Kuehne ◽  
Sun Jin Moon ◽  
...  

AbstractNanosensors have proven to be powerful tools to monitor single cells, achieving spatiotemporal precision even at molecular level. However, there has not been way of extending this approach to statistically relevant numbers of living cells. Herein, we design and fabricate nanosensor array in microfluidics that addresses this limitation, creating a Nanosensor Chemical Cytometry (NCC). nIR fluorescent carbon nanotube array is integrated along microfluidic channel through which flowing cells is guided. We can utilize the flowing cell itself as highly informative Gaussian lenses projecting nIR profiles and extract rich information. This unique biophotonic waveguide allows for quantified cross-correlation of biomolecular information with various physical properties and creates label-free chemical cytometer for cellular heterogeneity measurement. As an example, the NCC can profile the immune heterogeneities of human monocyte populations at attomolar sensitivity in completely non-destructive and real-time manner with rate of ~600 cells/hr, highest range demonstrated to date for state-of-the-art chemical cytometry.


Neuroforum ◽  
2019 ◽  
Vol 25 (3) ◽  
pp. 195-204
Author(s):  
Chotima Böttcher ◽  
Roman Sankowski ◽  
Josef Priller ◽  
Marco Prinz

Abstract The cellular composition of the central nervous system (CNS) is highly complex and dynamic. Regulation of this complexity is increasingly recognized to be spatially and temporally dependent during development, homeostasis and disease. Context-dependent cellular heterogeneity was shown for neuroectodermal cells as well as the myeloid compartment of the CNS. The brain myeloid compartment comprises microglia and other CNS-associated macrophages. These are brain-resident cells with critical roles in brain development, maintenance, and immune responses during states of disease. Profiling of CNS myeloid cell heterogeneity has been greatly facilitated in the past years by development of high-throughput technologies for single-cell analysis. This review summarizes current insights into heterogeneity of the CNS myeloid cell population determined by single-cell RNA sequencing and mass cytometry. The results offer invaluable insights into CNS biology and will facilitate the development of therapies for neurodegenerative and neuroinflammatory pathologies.


2019 ◽  
Vol 116 (36) ◽  
pp. 17957-17962 ◽  
Author(s):  
Su Bin Lim ◽  
Trifanny Yeo ◽  
Wen Di Lee ◽  
Ali Asgar S. Bhagat ◽  
Swee Jin Tan ◽  
...  

Despite pronounced genomic and transcriptomic heterogeneity in non–small-cell lung cancer (NSCLC) not only between tumors, but also within a tumor, validation of clinically relevant gene signatures for prognostication has relied upon single-tissue samples, including 2 commercially available multigene tests (MGTs). Here we report an unanticipated impact of intratumor heterogeneity (ITH) on risk prediction of recurrence in NSCLC, underscoring the need for a better genomic strategy to refine prognostication. By leveraging label-free, inertial-focusing microfluidic approaches in retrieving circulating tumor cells (CTCs) at single-cell resolution, we further identified specific gene signatures with distinct expression profiles in CTCs from patients with differing metastatic potential. Notably, a refined prognostic risk model that reconciles the level of ITH and CTC-derived gene expression data outperformed the initial classifier in predicting recurrence-free survival (RFS). We propose tailored approaches to providing reliable risk estimates while accounting for ITH-driven variance in NSCLC.


2019 ◽  
Vol 91 (15) ◽  
pp. 9777-9783 ◽  
Author(s):  
Huan Yao ◽  
Hansen Zhao ◽  
Xu Zhao ◽  
Xingyu Pan ◽  
Jiaxin Feng ◽  
...  

2022 ◽  
Author(s):  
Zizheng Shen ◽  
Hansen Zhao ◽  
Huan Yao ◽  
Xingyu Pan ◽  
Jinlei Yang ◽  
...  

Natural killer cell(NK cell)is an important immune cell which attracts increasing attention in cancer immunotherapy. Due to the heterogeneity of cells, individual cancer cell shows different resistance to NK cytotoxicity,...


2018 ◽  
Author(s):  
Subarna Palit ◽  
Fabian J. Theis ◽  
Christina E. Zielinski

AbstractRecent advances in cytometry have radically altered the fate of single-cell proteomics by allowing a more accurate understanding of complex biological systems. Mass cytometry (CyTOF) provides simultaneous single-cell measurements that are crucial to understand cellular heterogeneity and identify novel cellular subsets. High-dimensional CyTOF data were traditionally analyzed by gating on bivariate dot plots, which are not only laborious given the quadratic increase of complexity with dimension but are also biased through manual gating. This review aims to discuss the impact of new analysis techniques for in-depths insights into the dynamics of immune regulation obtained from static snapshot data and to provide tools to immunologists to address the high dimensionality of their single-cell data.


2015 ◽  
Vol 112 (28) ◽  
pp. E3661-E3668 ◽  
Author(s):  
Naside Gozde Durmus ◽  
H. Cumhur Tekin ◽  
Sinan Guven ◽  
Kaushik Sridhar ◽  
Ahu Arslan Yildiz ◽  
...  

Several cellular events cause permanent or transient changes in inherent magnetic and density properties of cells. Characterizing these changes in cell populations is crucial to understand cellular heterogeneity in cancer, immune response, infectious diseases, drug resistance, and evolution. Although magnetic levitation has previously been used for macroscale objects, its use in life sciences has been hindered by the inability to levitate microscale objects and by the toxicity of metal salts previously applied for levitation. Here, we use magnetic levitation principles for biological characterization and monitoring of cells and cellular events. We demonstrate that each cell type (i.e., cancer, blood, bacteria, and yeast) has a characteristic levitation profile, which we distinguish at an unprecedented resolution of 1 × 10−4g⋅mL−1. We have identified unique differences in levitation and density blueprints between breast, esophageal, colorectal, and nonsmall cell lung cancer cell lines, as well as heterogeneity within these seemingly homogenous cell populations. Furthermore, we demonstrate that changes in cellular density and levitation profiles can be monitored in real time at single-cell resolution, allowing quantification of heterogeneous temporal responses of each cell to environmental stressors. These data establish density as a powerful biomarker for investigating living systems and their responses. Thereby, our method enables rapid, density-based imaging and profiling of single cells with intriguing applications, such as label-free identification and monitoring of heterogeneous biological changes under various physiological conditions, including antibiotic or cancer treatment in personalized medicine.


2021 ◽  
Author(s):  
Juan Du ◽  
Junlei Zhang ◽  
Xiaocang Cao ◽  
Xun Wang ◽  
Jinyuan Song ◽  
...  

Abstract Background and Aims: Ulcerative colitis(UC) is a chronic inflammatory bowel disorder with highly cellular heterogeneity. Mass cytometry(Cy-TOF) and single-cell RNA sequencing (scRNA-seq) have revealed cellular heterogeneity of UC. However, comprehensive elucidation of tissue topological changes within the UC ecosystem is still missing. And we aimed to illustrate compositional and spatial changes of the UC ecosystem.Methods: Imaging mass cytometry (IMC) and scRNA-seq were applied to depict the single-cell landscape of colon ecosystem.Results: We noticed tissue topological changes featured with macrophage disappearance reaction (MDR) in UC region. MDR only occurred for CD163+ tissue-resident macrophages. We found reactive oxygen species (ROS) level were higher in UC region but ROS scavenging enzyme SOD1/2 were barely detected in resident macrophages, resulting selective oxidative protection for inflammatory macrophages and resident macrophage disappearance reaction. Furthermore, inflammatory macrophages replaced resident macrophages during UC, which played a key role in forming the inflammatory cellular network by producing TNF-α and IL-1β. Conclusions: Our study dissected the microenvironment of UC lesions at single-cell resolution while preserving its architecture, based on which, we discovered the mechanism of MDR in UC region and resident macrophage specific MDR resulted in infiltration of inflammatory macrophage, which formed the cytokine producing network within the local cellular neighborhood.


2020 ◽  
Author(s):  
Soo-Yeon Cho ◽  
Xun Gong ◽  
Volodymyr Koman ◽  
Matthias Kuehne ◽  
Sun Jin Moon ◽  
...  

Abstract Nanosensor have proven to be powerful tools to monitor single biological cells and organisms, achieving spatial and temporal precision even at the single molecule level. However, there has not been a way of extending this approach to statistically relevant numbers of living cells and organisms. Herein, we design and fabricate a high throughput nanosensor array in a microfluidic channel that addresses this limitation, creating a Nanosensor Chemical Cytometry (NCC). An array of nIR fluorescent single walled carbon nanotube (SWNT) nanosensors is integrated along a microfluidic channel through which a population of flowing cells is guided. We show that one can utilize the flowing cell itself as highly informative Gaussian lenses projecting nIR emission profiles and extract rich information on a per cell basis at high throughput. This unique biophotonic waveguide allows for quantified cross-correlation of the biomolecular information with physical properties such as cellular diameter, refractive index (RI), and eccentricity and creates a label-free chemical cytometer for the measurement of cellular heterogeneity with unprecedented precision. As an example, the NCC can profile the immune response heterogeneities of distinct human monocyte populations at attomolar (10-18 moles) sensitivity in a completely non-destructive and real-time manner with a rate of ~100 cells/frame, highest range demonstrated to date for state of the art chemical cytometry. We demonstrate distinct H2O2 efflux heterogeneities between 330 and 624 attomole/cell·min with cell projected areas between 271 and 263 µm2, eccentricity values between 0.405 and 0.363 and RI values between 1.383 and 1.377 for non-activated and activated human monocytes, respectively. Hence, we show that our nanotechnology based biophotonic cytometer has significant potential and versatility to answer important questions and provide new insight in immunology, cell manufacturing and biopharmaceutical research.


2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Yu-Sang Sabrina Yang ◽  
Prabhani U. Atukorale ◽  
Kelly D. Moynihan ◽  
Ahmet Bekdemir ◽  
Kavya Rakhra ◽  
...  

Abstract Inorganic nanoparticles (NPs) are studied as drug carriers, radiosensitizers and imaging agents, and characterizing nanoparticle biodistribution is essential for evaluating their efficacy and safety. Tracking NPs at the single-cell level with current technologies is complicated by the lack of reliable methods to stably label particles over extended durations in vivo. Here we demonstrate that mass cytometry by time-of-flight provides a label-free approach for inorganic nanoparticle quantitation in cells. Furthermore, mass cytometry can enumerate AuNPs with a lower detection limit of ∼10 AuNPs (3 nm core size) in a single cell with tandem multiparameter cellular phenotyping. Using the cellular distribution insights, we selected an amphiphilic surface ligand-coated AuNP that targeted myeloid dendritic cells in lymph nodes as a peptide antigen carrier, substantially increasing the efficacy of a model vaccine in a B16-OVA melanoma mouse model. This technology provides a powerful new level of insight into nanoparticle fate in vivo.


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