Erratum: High-throughput quantitation of inorganic nanoparticle biodistribution at the single-cell level using mass cytometry.
Ontology highlight
ABSTRACT:
SUBMITTER: Yang YS
PROVIDER: S-EPMC5561543 | biostudies-other | 2017 Aug
REPOSITORIES: biostudies-other
Similar Datasets
Project description: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.
| S-EPMC5247578 | biostudies-literature
Project description:Background: Mass cytometry (Cytometry by Time of Flight, CyTOF) allows single-cell characterization on the basis of specific metal-based cell markers. In addition, other metals in the mass range such as silver can be detected per cell. Bacteria are known to be sensible to silver and a protocol was developed to measure both the number of affected cells per population and the quantities of silver per cell. Methods: For mass cytometry ruthenium red was used as a marker for all cells of a population while parallel application of cisplatin discriminated live from dead cells. Silver quantities per cell and frequencies of silver containing cells in a population were measured by mass cytometry. In addition, live/dead subpopulations were analyzed by flow cytometry and distinguished by cell sorting based on ruthenium red and propidium iodide double staining. Verification of the cells' silver load was performed on the bulk level by using ICP-MS in combination with cell sorting. The protocol was developed by conveying both, fast and non-growing Pseudomonas putida cells as test organisms. Results: A workflow for labeling bacteria in order to be analyzed by mass cytometry was developed. Three different parameters were tested: ruthenium red provided counts for all bacterial cells in a population while consecutively applied cisplatin marked the frequency of dead cells. Apparent population heterogeneity was detected by different frequencies of silver containing cells. Silver quantities per cell were also well measurable. Generally, AgNP-10 treatment caused higher frequencies of dead cells, higher frequencies of silver containing cells and higher per-cell silver quantities. Due to an assumed chemical equilibrium of free and bound silver ions live and dead cells were associated with silver in equal quantities and this preferably during exponential growth. With ICP-MS up to 1.5 fg silver per bacterial cell were detected. Conclusion: An effective mass cytometry protocol was developed for the detection and quantification of silver in single bacterial cells of different physiological states. The silver quantities were generally heterogeneously distributed among cells in a population, the degree of which was dependent on micro-environmental conditions and on silver applied either in ion or nanoparticle-aggregated form.
| S-EPMC5511850 | biostudies-literature
Project description:Progress in the development and application of nanoengineered systems is limited by the availability of quantitative measurement techniques. For the engineering of nanoparticle (NP)-based systems, single NP characterization is essential, but existing methods are slow and low throughput. We demonstrate a flow spectroscopy technique capable of analyzing hundreds of nanoparticles per second and use this technique for the high throughput analysis of nanoparticle surface-enhanced resonant Raman scattering (SERRS) tags. By measuring Rayleigh and Raman scattering from thousands of individual tags, tag preparations can be characterized based on their brightness and uniformity. The rapid analysis of individual nanoparticles using high spectral resolution flow spectroscopy will be useful in many areas of nanoengineering.
| S-EPMC2730202 | biostudies-literature
Project description:IntroductionDiabetic ocular complications include sight-threatening consequences and decreased corneal sensitivity, characterized by decreased tear production, corneal sensitivity and delayed corneal epithelial wound healing. The pathogenesis of diabetic corneal disorders remains largely unknown. Growing evidence implies the participation of immune cells in the development of diabetic corneal diseases. Nonetheless, the immunological changes that result in diabetic corneal problems are largely unknown.MethodsMass cytometry by time of flight (CyTOF) was used to investigate immune cell cluster alterations associated with diabetic corneal disorders. CyTOF test was performed on corneal cells at a single level from 21-week-old diabetic (db/db) and non-diabetic (db/m) mice. A panel of 41 immune-related markers monitored different immune cell types in diabetic corneas. To investigate the proportion of each immune cell subpopulation, an unsupervised clustering method was employed, and T-distributed stochastic neighbor embedding was used to visualize the distinctions between different immune cell subsets.ResultsThrough CyTOF test, we identified 10 immune cell subsets in the corneal tissues. In a novel way, we discovered significant immune alterations in diabetic corneas, including pronounced alterations in T cells and myeloid cell subgroups in diabetic corneas linked to potential biomarkers, including CD103, CCR2, SiglecF, Ly6G, and CD172a. Comprehensive immunological profiling indicated remarkable changes in the immune microenvironment in diabetic corneas, characterized by a notable decrease in CD103+CD8+ tissue-resident memory T (TRM) cells and Tregs, as well as a dramatic increase of γδT cells and subsets of CD11b+Ly6G+ myeloid-derived suppressor cells (MDSCs).ConclusionCyTOF analysis revealed significant alterations in the immune microenvironment during the development of diabetic corneal complications. This study mapped the immune microenvironment landscape of type 2 diabetic corneas, providing a fundamental understanding of immune-driven diabetic corneal disorders.
| S-EPMC10507693 | biostudies-literature
Project description:This SuperSeries is composed of the SubSeries listed below.
2018-09-26 | GSE120446 | GEO
Project description:Haemostasis occurs at sites of vascular injury, where flowing blood forms a clot, a dynamic and heterogeneous fibrin-based biomaterial. Paramount in the clot's capability to stem haemorrhage are its changing mechanical properties, the major drivers of which are the contractile forces exerted by platelets against the fibrin scaffold. However, how platelets transduce microenvironmental cues to mediate contraction and alter clot mechanics is unknown. This is clinically relevant, as overly softened and stiffened clots are associated with bleeding and thrombotic disorders. Here, we report a high-throughput hydrogel-based platelet-contraction cytometer that quantifies single-platelet contraction forces in different clot microenvironments. We also show that platelets, via the Rho/ROCK pathway, synergistically couple mechanical and biochemical inputs to mediate contraction. Moreover, highly contractile platelet subpopulations present in healthy controls are conspicuously absent in a subset of patients with undiagnosed bleeding disorders, and therefore may function as a clinical diagnostic biophysical biomarker.
| S-EPMC5266633 | biostudies-literature
Project description:Fluorescent in situ hybridization (FISH) is a method that uses fluorescent probes to detect specific nucleic acid sequences at the single-cell level. Here we describe optimized protocols that exploit a highly sensitive FISH method based on branched DNA technology to detect mRNA and miRNA in human leukocytes. This technique can be multiplexed and combined with fluorescent antibody protein staining to address a variety of questions in heterogeneous cell populations. We demonstrate antigen-specific upregulation of IFNγ and IL-2 mRNAs in HIV- and CMV-specific T cells. We show simultaneous detection of cytokine mRNA and corresponding protein in single cells. We apply this method to detect mRNAs for which flow antibodies against the corresponding proteins are poor or are not available. We use this technique to show modulation of a microRNA critical for T-cell function, miR-155. We adapt this assay for simultaneous detection of mRNA and proteins by ImageStream technology.
| S-EPMC4256720 | biostudies-literature
Project description:In-depth organic mass cytometry reveals differential contents of 3-hydroxybutanoic acid on single cell level. Single-cell transcriptome indicates the expression difference of BHB downstream anti-oxidative stress proteins such as MT2A while Fluorescence Activated Cell Sorting (FACS) assay validates positive relationship between BHB and target proteins, which suggests that contents heterogeneity of BHB may endow cancer cells with variant ability to resist surrounding oxidative stress. Our ID-organic cytoMS paves the way for deep single-cell metabolome profiling and the investigation of cancer physiological and pathological processes. Comprehensive single-cell metabolic profiling is critical for revealing phenotypic heterogeneity and elucidating molecular mechanisms of biological processes. However, single-cell metabolomics remains challenging because of the limited metabolites coverage and disability of isomer discrimination. Herein, we establish a novel single-cell metabolomics platform of in-depth organic mass cytometry (ID-organic cytoMS). Extended single-cell analysis time guarantees sufficient MS/MS acquisition for metabolites identification and the isomers discrimination with online high-throughput analysis, achieving the largest number of about 600 metabolites identified in single cells. Fine sub-typing of MCF-7 cells are first demonstrated by differential contents of 3-hydroxybutanoic acid (BHB) among clusters. Single-cell transcriptome indicates the expression difference of BHB downstream anti-oxidative stress proteins such as MT2A while Fluorescence Activated Cell Sorting (FACS) assay validates positive relationship between BHB and target proteins, which suggests that contents heterogeneity of BHB may endow cancer cells with variant ability to resist surrounding oxidative stress. Our ID-organic cytoMS paves the way for deep single-cell metabolome profiling and the investigation of cancer physiological and pathological processes.
2024-03-31 | GSE262591 | GEO
Project description:Bulk RNA Sequencing of Healthy Bone Marrow Mononuclear Cells
2018-09-26 | GSE120444 | GEO
Project description:New techniques for single-cell analysis have led to insights into hematopoiesis and the immune system, but the ability of these techniques to cross-validate and reproducibly identify the biological variation in diverse human samples is currently unproven. We therefore performed a comprehensive assessment of human bone marrow cells using both single-cell RNA sequencing and multiparameter flow cytometry from twenty healthy adult human donors across a broad age range. These data characterize variation between healthy donors as well as age-associated changes in cell population frequencies. Direct comparison of techniques revealed discrepancy in the quantification of T lymphocyte and natural killer cell populations. Orthogonal validation of immunophenotyping using mass cytometry demonstrated good correlation with flow cytometry. Technical replicates using single-cell RNA sequencing matched robustly, while biological replicates showed variation. Given the increasing use of single-cell technologies in translational research, this resource serves as an important reference dataset and highlights opportunities for further refinement. [Funding source] Project Number: 1ZIAHL006163-05 Contact PI / Project Leader: HOURIGAN, CHRISTOPHER Title: DETECTION, PREVENTION AND TREATMENT OF ACUTE MYELOID LEUKEMIA (AML) RELAPSE. Awardee Organization: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
2018-09-21 | GSE120221 | GEO