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mass cytometry imaging

Worapol Ngamcherdtrakul, Thanapon Sangvanich, Shaun Goodyear, Moataz Reda, Shenda Gu, David J Castro, Primana Punnakitikashem, Wassana Yantasee
Multiparametric and high-content protein analysis of single cells or tissues cannot be accomplished with the currently available flow cytometry or imaging techniques utilizing fluorophore-labelled antibodies, because the number of spectrally resolvable fluorochromes is limited. In contrast, mass cytometry can resolve more signals by exploiting lanthanide-tagged antibodies; however, only about 100 metal reporters can be attached to an antibody molecule. This makes the sensitivity of lanthanide-tagged antibodies substantially lower than fluorescent reporters...
March 15, 2019: Bioengineering
Nicholas D Schmitt, Catherine M Rawlins, Elizabeth C Randall, Xianzhe Wang, Antonius Koller, Jared R Auclair, Jane-Marie Kowalski, Paul J Kowalski, Ed Luther, Alexander R Ivanov, Nathalie Y R Agar, Jeffrey N Agar
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) provides a unique in situ chemical profile that can include drugs, nucleic acids, metabolites, lipids, and proteins. MSI of individual cells (of a known cell type) affords a unique insight into normal and disease-related processes and is a prerequisite for combining the results of MSI and other single-cell modalities (e.g. mass cytometry and next-generation sequencing). Technological barriers have prevented the high-throughput assignment of MSI spectra from solid tissue preparations to their cell type...
March 6, 2019: Analytical Chemistry
Daniel E Carvajal-Hausdorf, Jonathan Patsenker, Kelly P Stanton, Franz Villarroel-Espindola, Amanda Esch, Ruth R Montgomery, Amanda Psyrri, Konstantine T Kalogeras, Vassiliki Kotoula, George Foutzilas, Kurt A Schalper, Yuval Kluger, David L Rimm
PURPOSE: Imaging Mass Cytometry (IMC) uses metal-conjugated antibodies to provide multidimensional, objective measurement of protein targets. We used this high-throughput platform to perform an 18-plex assessment of HER2 ICD/ECD, cytotoxic T cell infiltration and other structural and signaling proteins in a cohort of patients treated with trastuzumab to discover associations with trastuzumab benefit. EXPERIMENTAL DESIGN: An antibody panel for detection of 18 targets (Pancytokeratin, HER2 ICD, HER2 ECD, CD8, vimentin, cytokeratin 7, beta-catenin, HER3, MET, EGFR, ERK 1-2, MEK 1-2, PTEN, PI3K p110 alpha, Akt, mTOR, Ki67 and Histone H3) was used with a selection of trastuzumab-treated patients from the HeCOG 10/05 trial (n=180), and identified a case:control series...
February 22, 2019: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
James Turro, Pratiksha Singh, Manbeer Singh Sarao, Satish Tadepalli, Pramil Cheriyath
Background : Burkitt lymphoma is a rare, aggressive and rapidly fatal, B-cell non-Hodgkin's lymphoma. It has an incidence of 0.4/100,000 age-adjusted to the USA standard population. Here we describe the case of a 77-year-old patient who presented with Burkitt lymphoma. Case : A 77-year-old male presented to his primary care physician with fatigue and listlessness and was referred to the hospital with a white blood cell count (WBC)-23.7 K/uL (neutrophils 37%, lymphocyte 11%, blasts 9%) and platelets-19 K/uL...
2019: Journal of Community Hospital Internal Medicine Perspectives
Lin Ren, Masoumeh Dowlatshahi Pour, Per Malmberg, Andrew G Ewing
Micromolar concentration of zinc has been shown to significantly change the dynamics of exocytosis as well as the vesicle contents in a model cell line, providing direct evidence that zinc regulates neurotransmitter release. To provide insight about how zinc modulates these exocytotic processes, neurotransmitter release and vesicle content are now compared with single cell amperometry and intracellular impact vesicle cytometry at a range of zinc concentrations. Additionally, ToF-SIMS images of lipid distributions in the cell membrane after zinc treatment correlate to changes in exocytosis...
February 14, 2019: Chemistry: a European Journal
Zuzana Gajdosechova, Zoltan Mester
The need to assess the human and environmental risks of nanoparticles (NPs) has prompted an adaptation of existing techniques and the development of new ones. Nanoparticle analysis poses a great challenge as the analytical information has to consider both physical (e.g. size and shape) and chemical (e.g. elemental composition) state of the analyte. Furthermore, one has to contemplate the transformation of NPs during the sample preparation and provide sufficient information about the new species derived from such alteration...
February 14, 2019: Analytical and Bioanalytical Chemistry
Xiaoquan Rao, Shi Zhao, Zachary Braunstein, Hong Mao, Michael Razavi, Lihua Duan, Yingying Wei, Amelia C Toomey, Sanjay Rajagopalan, Jixin Zhong
BACKGROUND: We and others have shown that dipeptidyl peptidase-IV (DPP4) expression is increased in obesity/atherosclerosis and is positively correlated with atherosclerotic burden. However, the mechanism by which DPP4 expression is regulated in obesity remains unclear. In this study, we investigated the pathways regulating the expression of DPP4 on macrophages. METHODS: Flowsight® Imaging Flow Cytometry was employed for the detection of DPP4 and immunophenotyping...
February 6, 2019: EBioMedicine
Yue J Wang, Daniel Traum, Jonathan Schug, Long Gao, Chengyang Liu, Mark A Atkinson, Alvin C Powers, Michael D Feldman, Ali Naji, Kyong-Mi Chang, Klaus H Kaestner
The interaction between the immune system and endocrine cells in the pancreas is crucial for the initiation and progression of type 1 diabetes (T1D). Imaging mass cytometry (IMC) enables multiplexed assessment of the abundance and localization of more than 30 proteins on the same tissue section at 1-μm resolution. Herein, we have developed a panel of 33 antibodies that allows for the quantification of key cell types including pancreatic exocrine cells, islet cells, immune cells, and stromal components. We employed this panel to analyze 12 pancreata obtained from donors with clinically diagnosed T1D and 6 pancreata from non-diabetic controls...
January 28, 2019: Cell Metabolism
Nicolas Damond, Stefanie Engler, Vito R T Zanotelli, Denis Schapiro, Clive H Wasserfall, Irina Kusmartseva, Harry S Nick, Fabrizio Thorel, Pedro L Herrera, Mark A Atkinson, Bernd Bodenmiller
Type 1 diabetes (T1D) results from the autoimmune destruction of insulin-producing β cells. A comprehensive picture of the changes during T1D development is lacking due to limited sample availability, inability to sample longitudinally, and the paucity of technologies enabling comprehensive tissue profiling. Here, we analyzed 1,581 islets from 12 human donors, including eight with T1D, using imaging mass cytometry (IMC). IMC enabled simultaneous measurement of 35 biomarkers with single-cell and spatial resolution...
January 11, 2019: Cell Metabolism
Mimi Shin, Ying Wang, Jason R Borgus, B Jill Venton
Electrochemical measurements of neurotransmitters provide insight into the dynamics of neurotransmission. In this review, we describe the development of electrochemical measurements of neurotransmitters and how they started with extrasynaptic measurements but now are pushing toward synaptic measurements. Traditionally, behavioral measurements with biosensors or fast-scan cyclic voltammetry have monitored extrasynaptic levels of neurotransmitters, such as dopamine, serotonin, adenosine, glutamate, and acetylcholine...
February 1, 2019: Annual Review of Analytical Chemistry
Abdul Baset Abbas, Bingjing Lin, Chen Liu, Arwa Morshed, Jialiang Hu, Hanmei Xu
Immune-checkpoint blockades, suchas PD-1 monoclonal antibodies, have shown new promising avenues to treat cancers. Failure responsesof many cancer patients to these agents have led to a massive need for alternative strategies to optimize tumor immunotherapy. Currently, new therapeutic developments involve peptide blocking strategies, as they have high stability and low immunogenicity. Here, we have designed and synthesized a new peptide FITC-YT-16 to target PD-1. We have studied FITC-YT-16 by various experiments, including Molecular Operating Environment MOE modeling, purification testing by HPLC and LC mass, peptide/PD-1 conjugation and affinity by microscale thermophoresis (MST), and T cell immune-fluorescence imaging by fluorescence microscopy and flow cytometry...
January 29, 2019: International Journal of Molecular Sciences
Na Li, Vincent van Unen, Tamim Abdelaal, Nannan Guo, Sofya A Kasatskaya, Kristin Ladell, James E McLaren, Evgeny S Egorov, Mark Izraelson, Susana M Chuva de Sousa Lopes, Thomas Höllt, Olga V Britanova, Jeroen Eggermont, Noel F C C de Miranda, Dmitriy M Chudakov, David A Price, Boudewijn P F Lelieveldt, Frits Koning
The fetus is thought to be protected from exposure to foreign antigens, yet CD45RO+ T cells reside in the fetal intestine. Here we combined functional assays with mass cytometry, single-cell RNA sequencing and high-throughput T cell antigen receptor (TCR) sequencing to characterize the CD4+ T cell compartment in the human fetal intestine. We identified 22 CD4+ T cell clusters, including naive-like, regulatory-like and memory-like subpopulations, which were confirmed and further characterized at the transcriptional level...
January 21, 2019: Nature Immunology
Andrea K Pomerantz, Farid Sari-Sarraf, Kerri J Grove, Liliana Pedro, Patrick J Rudewicz, John W Fathman, Thomas Krucker
Single-cell imaging-based assays are an area of active and growing investment in drug discovery and development. This approach offers researchers the capability to interrogate rare subpopulations of cells with minimal sample consumption and multiplexed readouts. Recent technological advances in the optical interrogation and manipulation of single cells have substantially increased the throughput and sensitivity of these assays. Areas covered: In this review, the authors focus on three classes of single-cell imaging-based analyses: single-cell microscopy combined with microfluidics, mass spectrometric imaging for subcellular compound localization, and imaging mass cytometry (IMC)...
February 2019: Expert Opinion on Drug Discovery
Delphine C M Rolland, Megan S Lim, Kojo S J Elenitoba-Johnson
Mass spectrometry-based techniques now enable the unbiased identification of proteins in complex mixtures including proteins isolated from cells and tissues. These powerful tools permit near-complete annotation of proteins expressed in cells, tissues or organs. Further, these techniques permit the interrogation of the numerous posttranslational modifications that govern cell-specific responses to signaling cues and underlie the functional heterogeneity of cellular composition and contribute to biological complexity...
January 2019: Seminars in Hematology
Jérémie Decalf, Matthew L Albert, James Ziai
Tumor cell heterogeneity and tumor cell - stromal interactions are being explored as determinants of disease progression and treatment resistance in solid tumor and hematologic malignancies. As such, tools simultaneously capable of highly-multiplexed profiling of tissues' protein and RNA content, as well as interrogation of rare or single cells, are required to precisely characterize constituent tumor cell populations, infiltrating lymphocytes and stromal elements. Access to spatial relationships will enable more precise characterization of tumors, support patient stratification and may help to identify novel drug targets...
December 20, 2018: Journal of Pathology
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
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