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mass cytometry imaging

Allison N Schorzman, Andrew T Lucas, John R Kagel, William C Zamboni
Major advances in carrier-mediated agents (CMAs), which include nanoparticles, nanosomes, and conjugates, have revolutionized drug delivery capabilities over the past decade. While providing numerous advantages, such as greater solubility, duration of exposure, and delivery to the site of action over their small molecule counterparts, there is substantial variability in systemic clearance and distribution, tumor delivery, and pharmacologic effects (efficacy and toxicity) of these agents. In this chapter, we focus on the analytical and phenotypic methods required to design a study that characterizes the pharmacokinetics (PK) and pharmacodynamics (PD) of all forms of these nanoparticle-based drug agents...
2018: Methods in Molecular Biology
Julia Kh Leman, Sarah K Sandford, Janet L Rhodes, Roslyn A Kemp
Colorectal cancer (CRC) is a heterogeneous disease, with a diverse and plastic immune cell infiltrate. These immune cells play an important role in regulating tumour growth - progression or elimination. Some populations of cells have a strong correlation with disease-free survival, making them useful prognostic markers. In particular, the infiltrate of CD3+ and CD8+ T cells into CRC tumours has been validated worldwide as a valuable indicator of patient prognosis. However, the heterogeneity of the immune response, both between patients with tumours of different molecular subtypes, and within the tumour itself, necessitates the use of multiparametric analysis in the investigation of tumour-specific immune responses...
July 21, 2018: World Journal of Gastroenterology: WJG
Amélie St-Georges-Robillard, Mathieu Masse, Maxime Cahuzac, Mathias Strupler, Bishnubrata Patra, Adriana Mari Orimoto, Jennifer Kendall-Dupont, Benjamin Péant, Anne-Marie Mes-Masson, Frédéric Leblond, Thomas Gervais
Tumor spheroids represent a realistic 3D in vitro cancer model because they provide a missing link between monolayer cell culture and live tissues. While microfluidic chips can easily form and assay thousands of spheroids simultaneously, few commercial instruments are available to analyze this massive amount of data. Available techniques to measure spheroid response to external stimuli, such as confocal imaging and flow cytometry, are either not appropriate for 3D cultures, or destructive. We designed a wide-field hyperspectral imaging system to analyze multiple spheroids trapped in a microfluidic chip in a single acquisition...
August 6, 2018: Analyst
Alexey Y Karulin, Zoltán Megyesi, Richard Caspell, Jodi Hanson, Paul V Lehmann
Over the past decade, ELISPOT has become a highly implemented mainstream assay in immunological research, immune monitoring, and vaccine development. Unique single cell resolution along with high throughput potential sets ELISPOT apart from flow cytometry, ELISA, microarray- and bead-based multiplex assays. The necessity to unambiguously identify individual T and B cells that do, or do not co-express certain analytes, including polyfunctional cytokine producing T cells has stimulated the development of multi-color ELISPOT assays...
2018: Methods in Molecular Biology
Jozef I Nissimov, Rebecca Vandzura, Christopher T Johns, Frank Natale, Liti Haramaty, Kay D Bidle
Emiliania huxleyi produces calcium carbonate (CaCO3 ) coccoliths and transparent exopolymer particles (TEP), sticky, acidic carbohydrates that facilitate aggregation. E. huxleyi's extensive oceanic blooms are often terminated by coccolithoviruses (EhVs) with the transport of cellular debris and associated particulate organic carbon (POC) to depth being facilitated by TEP-bound "marine snow" aggregates. The dynamics of TEP production and particle aggregation in response to EhV infection are poorly understood...
June 19, 2018: Environmental Microbiology
Giulio Caracciolo, Sara Palchetti, Luca Digiacomo, Riccardo Zene Zini Chiozzi, Anna Laura Capriotti, Heinz Amenitsch, Paolo Maria Tentori, Valentina Palmieri, Massimiliano Papi, Francesco Cardarelli, Daniela Pozzi, Aldo Laganà
More than 20 years after its approval by the Food and Drug Administration (FDA), liposomal doxorubicin (DOX) is still the drug of choice for the treatment of breast cancer and other conditions such as ovarian cancer and multiple myeloma. Yet, despite the efforts, liposomal DOX did not satisfy expectations at the clinical level. When liposomal drugs enter a physiological environment, their surface gets coated by a dynamic biomolecular corona (BC). The BC changes liposome's synthetic identity, providing it with a new one, referred to as "biological identity" (size, aggregation state, and BC composition)...
July 11, 2018: ACS Applied Materials & Interfaces
Xiaoying Ye, Brian T Luke, Bih-Rong Wei, Jan A Kaczmarczyk, Jadranka Loncarek, Jennifer E Dwyer, Donald J Johann, Richard G Saul, Dwight V Nissley, Frank McCormick, Gordon R Whiteley, Josip Blonder
The most widely used cancer animal model is the human-murine tumor xenograft. Unbiased molecular dissection of tumor parenchyma versus stroma in human-murine xenografts is critical for elucidating dysregulated protein networks/pathways and developing therapeutics that may target these two functionally codependent compartments. Although antibody-reliant technologies (e.g., immunohistochemistry, imaging mass cytometry) are capable of distinguishing tumor-proper versus stromal proteins, the breadth or extent of targets is limited...
May 29, 2018: Oncotarget
Deon B Doxie, Allison R Greenplate, Jocelyn S Gandelman, Kirsten E Diggins, Caroline E Roe, Kimberly B Dahlman, Jeffrey A Sosman, Mark C Kelley, Jonathan M Irish
Little is known about the in vivo impacts of targeted therapy on melanoma cell abundance and protein expression. Here, 21 antibodies were added to an established melanoma mass cytometry panel to measure 32 cellular features, distinguish malignant cells, and characterize dabrafenib and trametinib responses in BRAFV600mut melanoma. Tumor cells were biopsied before neoadjuvant therapy and compared to cells surgically resected from the same site after 4 weeks of therapy. Approximately 50,000 cells per tumor were characterized by mass cytometry and computational tools t-SNE/viSNE, FlowSOM, and MEM...
May 19, 2018: Pigment Cell & Melanoma Research
Federico Boschi, Francesco De Sanctis, Stefano Ugel, Antonello E Spinelli
Cancer immunotherapy is a promising strategy based on the ability of the immune system to kill selected cells. In the development of an effective T-cell therapy, the noninvasive cell tracking methods play a crucial role. Here, we investigate the potentialities of T-cell marked with radionuclides in order to detect their localization with imaging techniques in small animal rodents. A protocol to label T-cells with 32 P-ATP was tested and evaluated. The homing of 32 P-ATP labeled T lymphocytes was investigated by Cerenkov luminescence imaging (CLI) and radioluminescence imaging (RLI)...
May 16, 2018: Journal of Biophotonics
Sara Khademi, Saeed Sarkar, Ali Shakeri-Zadeh, Neda Attaran, Sharmin Kharrazi, Mohammad Reza Ay, Hossein Ghadiri
Development of various cost-effective multifunctional nanoprobes for efficient targeted molecular imaging of tumors remains a great challenge in medicine. Herein, we report a simple method of forming folic acid-targeted multifunctional gold nanoparticles via cost-effective cysteamine as a template for tumor molecular computed tomography (CT) imaging technique. The formed multifunctional cysteamine-folic acid conjugated gold nanoparticles (FA-Cys-AuNPs) were characterized via different techniques. Colony assay, hematoxylin and eosin (H&E), MTT, and flow cytometry analysis were used to evaluate the cytocompatibility of the particles...
August 1, 2018: Materials Science & Engineering. C, Materials for Biological Applications
Bedilu Allo, Xudong Lou, Alexandre Bouzekri, Olga Ornatsky
Mass cytometry is a highly multiplexed single-cell analysis platform that uses metal-tagged reagents to identify multiple cellular biomarkers. The current metal-tagged reagent preparation employs thiol-maleimide chemistry to covalently couple maleimide-functionalized metal-chelating polymers (MCPs) with antibodies (Abs), a process that requires partial reduction of the Ab to form reactive thiol groups. However, some classes of Abs (for example, IgM) as well as biomolecules lacking cysteine residues have been challenging to label using this method...
June 20, 2018: Bioconjugate Chemistry
Daniel Schulz, Vito Riccardo Tomaso Zanotelli, Jana Raja Fischer, Denis Schapiro, Stefanie Engler, Xiao-Kang Lun, Hartland Warren Jackson, Bernd Bodenmiller
No abstract text is available yet for this article.
April 25, 2018: Cell Systems
Rufael Chekol, Viswas Raja Solomon, Elahe Alizadeh, Wendy Bernhard, Darrell Fisher, Wayne Hill, Kris Barreto, John Francis DeCoteau, Angel Casaco Parada, Clarence Ronald Geyer, Humphrey Fonge
Rationale: Epidermal growth factor receptor (EGFR) upregulation is associated with enhanced proliferation and drug resistance in a number of cancers. Nimotuzumab is a humanized monoclonal antibody with high affinity for EGFR. The objective of this study was to determine if 89 Zr-DFO-nimotuzumab could be suitable for human use as a PET probe for quantifying EGFR in vivo . Methods: To evaluate the pharmacokinetics, biodistribution, microPET imaging, radiation dosimetry, and normal tissue toxicity in tumor and non-tumor bearing mice of 89 Zr-desferoxamine-nimotuzumab (89 Zr-DFO-nimotuzumab) of a product prepared under GMP conditions...
March 30, 2018: Oncotarget
Wenhai Lin, Lei Yin, Tingting Sun, Tingting Wang, Zhigang Xie, Jingkai Gu, Xiabin Jing
Fundamental studies on the cellular uptake and drug release of PEGylated nanomedicines are beneficial to understand their fate in vivo and construct ideal nanoparticle formulations. In this work, the detailed metabolic process of PEGylated doxorubicin (Dox) nanomedicines were investigated via confocal laser scanning microscopy (CLSM), flow cytometry (FCM), cytotoxicity test, fluorescence imaging in vivo (FLIV) and liquid chromatography tandem mass spectrometry (LC-MS/MS). Among them, only LC-MS/MS could accurately determine the content of PEGylated Dox and Dox in vitro and in vivo...
May 14, 2018: Biomacromolecules
Stéphane Chevrier, Helena L Crowell, Vito R T Zanotelli, Stefanie Engler, Mark D Robinson, Bernd Bodenmiller
The advent of mass cytometry increased the number of parameters measured at the single-cell level while decreasing the extent of crosstalk between channels relative to dye-based flow cytometry. Although reduced, spillover still exists in mass cytometry data, and minimizing its effect requires considerable expert knowledge and substantial experimental effort. Here, we describe a novel bead-based compensation workflow and R-based software that estimates and corrects for interference between channels. We performed an in-depth characterization of the spillover properties in mass cytometry, including limitations defined by the linear range of the mass cytometer and the reproducibility of the spillover over time and across machines...
May 23, 2018: Cell Systems
F A Escobar, L Pantanowitz, J L Picarsic, F E Craig, J P Simons, P A Viswanathan, S Yilmaz, S E Monaco
INTRODUCTION: Ectopic thymic tissue can arise as an asymptomatic neck mass, which may be detected on imaging studies. The aim of this study was to determine the incidence of ectopic thymic tissue in paediatric FNAs and to the correlate clinical, radiological and cytomorphological findings. METHODS: FNAs in children with neck and mediastinal lesions performed between January 2012 and July 2016 were reviewed for cases of ectopic thymus. These were then evaluated and correlated with the cytology findings...
June 2018: Cytopathology: Official Journal of the British Society for Clinical Cytology
Chi Ching Goh, Maximilien Evrard, Shu Zhen Chong, Yingrou Tan, Leonard De Li Tan, Karen Wei Weng Teng, Wolfgang Weninger, David Laurence Becker, Hong Liang Tey, Evan William Newell, Bin Liu, Lai Guan Ng
Pressure ulcers are a chronic problem for patients or the elderly who require extended periods of bed rest. The formation of ulcers is due to repeated cycles of ischemia-reperfusion (IR), which initiates an inflammatory response. Advanced ulcers disrupt the skin barrier, resulting in further complications. To date, the immunological aspect of skin IR has been understudied, partly due to the complexity of the skin immune cells. Through a combination of mass cytometry, confocal imaging and intravital multiphoton imaging, this study establishes a workflow for multidimensionality single cell analysis of skin myeloid cell responses in the context of IR injury with high spatiotemporal resolution...
June 2018: European Journal of Immunology
Tess Melinda Brodie, Vinko Tosevski
Mass cytometry (cytometry by time-of-flight, CyTOF) is a high-dimensional single-cell analytical technology that allows for highly multiplexed measurements of protein or nucleic acid abundances by bringing together the detection capacity of atomic mass spectroscopy and the sample preparation workflow typical of regular flow cytometry. In 2014 the mass cytometer was adapted for the acquisition of samples from microscopy slides (termed imaging mass cytometry), greatly increasing the applicability of this technology with the inclusion of spatial information...
2018: Methods in Molecular Biology
Qingqing Meng, Aiping Wang, Hongchen Hua, Ying Jiang, Yiyun Wang, Hongjie Mu, Zimei Wu, Kaoxiang Sun
Background: Safe and effective delivery of therapeutic drugs to the brain is important for successful therapy of Alzheimer's disease (AD). Purpose: To develop Huperzine A (HupA)-loaded, mucoadhesive and targeted polylactide-co-glycoside (PLGA) nanoparticles (NPs) with surface modification by lactoferrin (Lf)-conjugated N-trimethylated chitosan (TMC) (HupA Lf-TMC NPs) for efficient intranasal delivery of HupA to the brain for AD treatment. Methods: HupA Lf-TMC NPs were prepared using the emulsion-solvent evaporation method and optimized using the Box-Behnken design...
2018: International Journal of Nanomedicine
Liron Pantanowitz, Frederic Preffer, David C Wilbur
Novel techniques have been developed to image cells at cellular and subcellular levels. They allow images to be analyzed with ultra-high resolution, in 2D and/or 3D. Several of these tools have been tested on cytology specimens demonstrating emerging applications that are likely to change the field of cytopathology. This review covers several of these advanced imaging methods. The use of optical coherence tomography to perform optical biopsies during endoscopic ultrasound procedures or visualize cells within effusion samples is discussed...
February 12, 2018: Diagnostic Cytopathology
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