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mass cytometry imaging

Chandresh R Gajera, Rosemary Fernandez, Nadia Postupna, Kathleen S Montine, Edward J Fox, Dmitry Tebaykin, Michael Angelo, Sean C Bendall, C Dirk Keene, Thomas J Montine
BACKGROUND: Synaptic alterations, especially presynaptic changes, are cardinal features of neurodegenerative diseases and strongly correlate with cognitive decline. NEW METHOD: We report "Mass Synaptometry" for the high-dimensional analysis of individual human synaptosomes, enriched nerve terminals from brain. This method was adapted from cytometry by time-of-flight mass spectrometry (CyTOF), which is commonly used for single-cell analysis of immune and blood cells...
November 20, 2018: Journal of Neuroscience Methods
Ponarulselvam Sekar, Duen-Yi Huang, Shie-Liang Hsieh, Shwu-Fen Chang, Wan-Wan Lin
BACKGROUND: P2X7 is ubiquitously expressed in myeloid cells and regulates the pathophysiology of inflammatory diseases. Since mitochondrial function in microglia is highly associated with microglial functions in controlling neuronal plasticity and brain homeostasis, we interested to explore the roles of P2X7 in mitochondrial and lysosomal functions as well as mitophagy in microglia. METHODS: P2X7-/- bone marrow-derived macrophages (BMDM), primary microglia and BV-2 immortalized microglial cells were used to detect the particular protein expression by immunoblotting...
November 20, 2018: Cell Communication and Signaling: CCS
G P Vishnu Vardhan, M Hema, C Sushmitha, H S Savithri, Usha Natraj, M R N Murthy
The capsids of viruses have a high degree of symmetry. Therefore, virus nanoparticles (VNPs) can be programmed to display many imaging agents precisely. Plant VNPs are biocompatible, biodegradable and non-infectious to mammals. We have carried out bioconjugation of sesbania mosaic virus (SeMV), a well characterized plant virus, with fluorophores using reactive lysine-N-hydroxysuccinimide ester and cysteine-maleimide chemistries. Monitoring of cellular internalization of labelled SeMV nanoparticles (NPs) by confocal microscopy and flow cytometry showed that the particles have a natural preference for entry into MDA-MB-231 (breast cancer) cells, although they could also enter various other cell lines...
November 14, 2018: Archives of Virology
Joshua Whited, Czharena Kay Rama, Xue-Long Sun
Glycan-binding molecules, such as lectins, are very important tools for characterizing, imaging, or targeting glycans and are often involved in either physiological or pathological processes. However, their availability is far less compared to the diversity of native glycans. Therefore, development of lectin mimetics with desired specificity and affinity is in high demand. Boronic acid reacts with 1,2- and 1,3-diols of saccharides in aqueous media through reversible boronate ester formation and are regarded as synthetic lectin mimetics...
October 31, 2018: ACS Omega
K Lemdani, H Salmon, R Gahoual, M Bessodes, D Scherman, P Houzé, J Seguin, N Mignet
In the context of increasing liver diseases, no contrast agent is currently available in Europe and the United States to directly assess the liver function. Only neolactosylated human serum albumin is being clinically used in Asia. In order to perform preclinical studies in the context of liver diseases, we conceived a fluorescent lactosylated albumin for the quantification of liver functional cells (l-Cyal). Precise characterization was achieved in order to determine the amounts of lactose and Cyanine 5 (Cy5) coupled to the albumin...
November 8, 2018: Nanoscale
Khursheed Ali, Bilal Ahmed, Mohammad Saghir Khan, Javed Musarrat
Biogenic hematite (α-Fe2 O3 ) nanoparticles (NPs) of average size <10 nm were synthesized using green approach with Aloe vera extract (ALE). The aim of the study was to assess the protective effect of extracellular polymeric substances (EPS) against antibacterial and antibiofilm activities of ALE-α-Fe2 O3 NPs in normal EPS producers (pristine) and experimentally modified (low-EPS) Pseudomonas aeruginosa (P. aeruginosa) cells and the mechanism of cell killing. Formation of ALE-α-Fe2 O3 NPs has been validated by X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM) and Fourier-transformed infrared spectroscopy (FTIR) analysis...
November 2018: Journal of Photochemistry and Photobiology. B, Biology
Yuan Zhao, Mohamed Uduman, Jacqueline H Y Siu, Thomas J Tull, Jeremy D Sanderson, Yu-Chang Bryan Wu, Julian Q Zhou, Nedyalko Petrov, Richard Ellis, Katrina Todd, Konstantia-Maria Chavele, William Guesdon, Anna Vossenkamper, Wayel Jassem, David P D'Cruz, David J Fear, Susan John, Dagmar Scheel-Toellner, Claire Hopkins, Estefania Moreno, Natalie L Woodman, Francesca Ciccarelli, Susanne Heck, Steven H Kleinstein, Mats Bemark, Jo Spencer
Human memory B cells and marginal zone (MZ) B cells share common features such as the expression of CD27 and somatic mutations in their IGHV and BCL6 genes, but the relationship between them is controversial. Here, we show phenotypic progression within lymphoid tissues as MZ B cells emerge from the mature naïve B cell pool via a precursor CD27- CD45RBMEM55+ population distant from memory cells. By imaging mass cytometry, we find that MZ B cells and memory B cells occupy different microanatomical niches in organised gut lymphoid tissues...
September 21, 2018: Nature Communications
Marieke E Ijsselsteijn, Thomas P Brouwer, Ziena Abdulrahman, Eileen Reidy, Ana Ramalheiro, A Marijne Heeren, Alexander Vahrmeijer, Ekaterina S Jordanova, Noel F C C de Miranda
Checkpoint blockade immunotherapies have revolutionised cancer treatment in the last decade. Nevertheless, these are only beneficial for a small proportion of cancer patients. Important prognosticators for response to immunotherapy are the mutation burden of tumours as well as the quality and quantity of tumour-infiltrating immune cells. High-throughput multiplex immunophenotyping technologies have a central role in deciphering the complexity of anti-tumour immune responses. Current techniques for the immunophenotyping of solid tumours are held back by the lack of spatial context, limitations in the number of targets that can be visualised simultaneously, and/or cumbersome protocols...
September 7, 2018: Journal of Pathology. Clinical Research
Tiffany Porta Siegel, Gregory Hamm, Josephine Bunch, Jo Cappell, John S Fletcher, Kristina Schwamborn
Over the last two decades, mass spectrometry imaging (MSI) has been increasingly employed to investigate the spatial distribution of a wide variety of molecules in complex biological samples. MSI has demonstrated its potential in numerous applications from drug discovery, disease state evaluation through proteomic and/or metabolomic studies. Significant technological and methodological advancements have addressed natural limitations of the techniques, i.e., increased spatial resolution, increased detection sensitivity especially for large molecules, higher throughput analysis and data management...
December 2018: Molecular Imaging and Biology: MIB: the Official Publication of the Academy of Molecular Imaging
Kuo-Ching Mei, Artur Ghazaryan, Er Zhen Teoh, Huw D Summers, Yueting Li, Belén Ballesteros, Justyna Piasecka, Adam Walters, Robert C Hider, Volker Mailänder, Khuloud T Al-Jamal
Hard corona (HC) protein, i.e., the environmental proteins of the biological medium that are bound to a nanosurface, is known to affect the biological fate of a nanomedicine. Due to the size, curvature, and specific surface area (SSA) 3-factor interactions inherited in the traditional 3D nanoparticle, HC-dependent bio-nano interactions are often poorly probed and interpreted. Here, the first HC-by-design case study in 2D is demonstrated that sequentially and linearly changes the HC quantity using functionalized graphene oxide (GO) nanosheets...
August 24, 2018: Advanced Materials
Yanshuo Cao, Qing Chang, Michael Cabanero, Wenjiang Zhang, Sara Hafezi-Bakhtiari, David Hedley, Gail Darling, Fayez Quereshy, Raymond Jang, Elena Elimova, Jennifer Knox, Olga Ornatsky, Stefano Serra, Eric Chen
PURPOSE: There is a wide range in tumor response following preoperative chemotherapy in locally advanced gastric or gastroesophageal junction cancers. We investigated the relationship between tumor platinum levels and pathological responses in these patients. METHODS: Tumor and adjacent normal tissues were retrieved. Pathological responses were assessed per standard criteria. Tissue platinum concentrations were determined with high-performance liquid chromatography mass spectrometry...
August 16, 2018: Journal of Gastrointestinal Cancer
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...
November 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
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