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Cytometry. Part A: the Journal of the International Society for Analytical Cytology

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https://www.readbyqxmd.com/read/29220555/multiparameter-cytometric-analysis-of-complex-cellular-response
#1
Šárka Šimečková, Radek Fedr, Ján Remšík, Zuzana Kahounová, Eva Slabáková, Karel Souček
Complex analysis of cellular responses after experimental treatment is important for screening, mechanistic understanding of treatment effects, and the identification of sensitive and resistant cell phenotypes. Modern multicolor flow cytometry has demonstrated its power for such analyses. Here, we introduce a multiparametric protocol for complex analysis of cytokinetics by the simultaneous detection of seven fluorescence parameters. This analysis includes the detection of two surface markers for immunophenotyping, analysis of proliferation based on the cell cycle and the measurement of incorporated nucleoside analogue 5-ethynyl-2'-deoxyuridine (EdU) in newly synthesized DNA, analysis of DNA damage using an anti-phospho-histone H2A...
December 8, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29205767/atomic-mass-tag-of-bismuth-209-for-increasing-the-immunoassay-multiplexing-capacity-of-mass-cytometry
#2
Guojun Han, Shih-Yu Chen, Veronica D Gonzalez, Eli R Zunder, Wendy J Fantl, Garry P Nolan
Mass cytometry (or CyTOF) is an atomic mass spectrometry-based single-cell immunoassay technology, which has provided an increasingly systematic and sophisticated view in basic biological and clinical studies. Using elemental reporters composed of stable heavy metal isotopes, more than 50 cellular parameters are measured simultaneously. However, this current multiplexing does not meet the theoretical capability of CyTOF instrumentation with 135 detectable channels, primarily due to the limitation of available chemistries for conjugating elemental mass tags to affinity reagents...
December 4, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29194963/simultaneous-detection-of-protein-and-mrna-in-jurkat-and-kg-1a-cells-by-mass-cytometry
#3
Anastasia Mavropoulos, Bedilu Allo, Mingxiao He, Emily Park, Daniel Majonis, Olga Ornatsky
Mass cytometry uniquely enables high-dimensional single-cell analysis of complex populations. This recently developed technology is based on inductively coupled time-of-flight mass spectrometry for multiplex proteomic analysis of more than 40 markers per cell. The ability to characterize the transcriptome is critical for the understanding of disease pathophysiology, medical diagnostics, and drug discovery. Current techniques allowing the in situ detection of transcripts in single cells are limited to a small number of simultaneous targets and are generally tedious and labor-intensive...
November 30, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29194951/quantification-of-large-scale-dna-organization-for-predicting-prostate-cancer-recurrence
#4
Calum MacAulay, Mira Keyes, Malcolm Hayes, Andrea Lo, Gang Wang, Martial Guillaud, Martin Gleave, Laden Fazli, Jagoda Korbelik, Colin Collins, Sarah Keyes, Branko Palcic
This study investigates whether Genomic Organization at Large Scales (which we propose to call GOALS) as quantified via nuclear phenotype characteristics and cell sociology features (describing cell organization within tissue) collected from prostate tissue microarrays (TMAs) can separate biochemical failure from biochemical nonevidence of disease (BNED) after radical prostatectomy (RP). Of the 78 prostate cancer tissue cores collected from patients treated with RP, 16 who developed biochemical relapse (failure group) and 16 who were BNED patients (nonfailure group) were included in the analyses (36 cores from 32 patients)...
November 30, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29165907/stripping-flow-cytometry-how-many-detectors-do-we-need-for-bacterial-identification
#5
Peter Rubbens, Ruben Props, Cristina Garcia-Timermans, Nico Boon, Willem Waegeman
Multicolor approaches are challenging for microbial flow cytometry; as flow cytometers are mainly developed for biomedical applications, modern instruments contain more detectors than needed. Some of these additional fluorescence detectors measure biological information due to spectral overlap, yet the extent to which this information is relevant for the identification of bacterial populations is ambiguous. In this paper we characterize the usefulness of these additional detectors. We propose a data-driven detector selection method to select the smallest subset of detectors that will optimally discriminate between bacterial populations...
November 22, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29165899/quantitative-performance-evaluation-of-a-back-illuminated-scmos-camera-with-95-qe-for-super-resolution-localization-microscopy
#6
Yujie Wang, Lingxi Zhao, Zhe Hu, Yina Wang, Zeyu Zhao, Luchang Li, Zhen-Li Huang
Scientific Complementary Metal Oxide Semiconductor (sCMOS) cameras were introduced into the market in 2009 and are now becoming a major type of commercial cameras for low-light imaging. sCMOS cameras provide simultaneously low read noise, high readout speed, and large pixel array; however, the relatively low quantum efficiency (QE) of sCMOS cameras has been a major limitation for its application in single molecule imaging, especially super-resolution localization microscopy which requires high detection sensitivity...
November 22, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29156109/a-microfabricated-96-well-wound-healing-assay
#7
Shaoliang Luan, Rui Hao, Yuanchen Wei, Deyong Chen, Beiyuan Fan, Fengliang Dong, Wei Guo, Junbo Wang, Jian Chen
This article presents a microfabricated 96-well wound-healing assay enabling high-throughput measurement of cellular migration capabilities. Within each well, the middle area is the wound region, made of microfabricated gold surface with self-assembled PEG repellent for cell seeding. After the formation of a cellular confluent monolayer around the wound region, collagen solution was applied to form three-dimensional matrix to cover the PEG surface, initiating the wound-healing process. By interpreting the numbers of migrated cells into the wound regions as a function of specific stimuli with different concentrations, EC50 (half-maximal effective concentration) was obtained...
November 20, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29125897/image-analysis-of-neural-stem-cell-division-patterns-in-the-zebrafish-brain
#8
Valerio Lupperger, Felix Buggenthin, Prisca Chapouton, Carsten Marr
Proliferating stem cells in the adult body are the source of constant regeneration. In the brain, neural stem cells (NSCs) divide to maintain the stem cell population and generate neural progenitor cells that eventually replenish mature neurons and glial cells. How much spatial coordination of NSC division and differentiation is present in a functional brain is an open question. To quantify the patterns of stem cell divisions, one has to (i) identify the pool of NSCs that have the ability to divide, (ii) determine NSCs that divide within a given time window, and (iii) analyze the degree of spatial coordination...
November 10, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29077263/cell-shape-characterization-and-classification-with-discrete-fourier-transforms-and-self-organizing-maps
#9
Fabian L Kriegel, Ralf Köhler, Jannike Bayat-Sarmadi, Simon Bayerl, Anja E Hauser, Raluca Niesner, Andreas Luch, Zoltan Cseresnyes
Cells in their natural environment often exhibit complex kinetic behavior and radical adjustments of their shapes. This enables them to accommodate to short- and long-term changes in their surroundings under physiological and pathological conditions. Intravital multi-photon microscopy is a powerful tool to record this complex behavior. Traditionally, cell behavior is characterized by tracking the cells' movements, which yields numerous parameters describing the spatiotemporal characteristics of cells. Cells can be classified according to their tracking behavior using all or a subset of these kinetic parameters...
October 27, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29072818/isolation-cultivation-and-characterization-of-human-mesenchymal-stem-cells
#10
REVIEW
Dolly Mushahary, Andreas Spittler, Cornelia Kasper, Viktoria Weber, Verena Charwat
Mesenchymal stem cells (MSC) exhibit a high self-renewal capacity, multilineage differentiation potential and immunomodulatory properties. This set of exceptional features makes them an attractive tool for research and clinical application. However, MSC are far from being a uniform cell type, which makes standardization difficult. The exact properties of human MSC (hMSC) can vary greatly depending on multiple parameters including tissue source, isolation method and medium composition. In this review we address the most important influence factors...
October 26, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/29030991/omip-041-optimized-multicolor-immunofluorescence-panel-rat-microglial-staining-protocol
#11
Naama E Toledano Furman, Karthik S Prabhakara, Supinder Bedi, Charles S Cox, Scott D Olson
No abstract text is available yet for this article.
October 14, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28976646/untangling-cell-tracks-quantifying-cell-migration-by-time-lapse-image-data-analysis
#12
REVIEW
Carl-Magnus Svensson, Anna Medyukhina, Ivan Belyaev, Naim Al-Zaben, Marc Thilo Figge
Automated microscopy has given researchers access to great amounts of live cell imaging data from in vitro and in vivo experiments. Much focus has been put on extracting cell tracks from such data using a plethora of segmentation and tracking algorithms, but further analysis is normally required to draw biologically relevant conclusions. Such relevant conclusions may be whether the migration is directed or not, whether the population has homogeneous or heterogeneous migration patterns. This review focuses on the analysis of cell migration data that are extracted from time lapse images...
October 4, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28976638/bivariate-flow-cytometric-analysis-and-sorting-of-different-types-of-maize-starch-grains
#13
Xudong Zhang, Jiaojiao Feng, Heng Wang, Jianchu Zhu, Yuyue Zhong, Linsan Liu, Shutu Xu, Renhe Zhang, Xinghua Zhang, Jiquan Xue, Dongwei Guo
Particle-size distribution, granular structure, and composition significantly affect the physicochemical properties, rheological properties, and nutritional function of starch. Flow cytometry and flow sorting are widely considered convenient and efficient ways of classifying and separating natural biological particles or other substances into subpopulations, respectively, based on the differential response of each component to stimulation by a light beam; the results allow for the correlation analysis of parameters...
October 4, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28945315/quantitating-adcc-against-adherent-cells-impedance-based-detection-is-superior-to-release-membrane-permeability-or-caspase-activation-assays-in-resolving-antibody-dose-response
#14
Gábor Tóth, János Szöllősi, György Vereb
Monoclonal antibody-based immunotherapeutics will dominate Pharma's next generation of blockbuster drugs, and Fc-associated functions, including antibody dependent cellular cytotoxicity (ADCC) are among the highly desired activities mediated by these antibodies. Therefore, quantitative evaluation of ADCC is required during drug development. Our objective was to find the most suitable and reliable nonradioactive method for quantitative analysis of in vitro ADCC against adherent cells, which often serve as models for solid tumors...
September 25, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28940935/rapid-comparative-immunophenotyping-of-human-mesenchymal-stromal-cells-by-a-modified-fluorescent-cell-barcoding-flow-cytometric-assay
#15
Tamara Lekishvili, Jonathan J Campbell
Flow cytometry immunophenotyping is a sensitive technique allowing rapid characterization of single cells within heterogeneous populations, but it includes several subjective steps during sample analysis that impact the development of standardized methodology. Proposed strategies to overcome these limitations include fluorescent cell barcoding (FCB), which facilitates multiplexed sample evaluation with increased data reproducibility whilst reducing labeling variation, materials, and time. To date, the FCB assay has found utility for analyzing the phosphorylation status of intracellular targets but has not been intensively employed for cellular immunophenotypic analyses using cell surface markers...
September 22, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28941170/imaging-flow-cytometry-assays-for-quantifying-pigment-grade-titanium-dioxide-particle-internalization-and-interactions-with-immune-cells-in-whole-blood
#16
Rachel E Hewitt, Bradley Vis, Laetitia C Pele, Nuno Faria, Jonathan J Powell
Pigment grade titanium dioxide is composed of sub-micron sized particles, including a nanofraction, and is widely utilized in food, cosmetic, pharmaceutical, and biomedical industries. Oral exposure to pigment grade titanium dioxide results in at least some material entering the circulation in humans, although subsequent interactions with blood immune cells are unknown. Pigment grade titanium dioxide is employed for its strong light scattering properties, and this work exploited that attribute to determine whether single cell-particle associations could be determined in immune cells of human whole blood at "real life" concentrations...
September 20, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28941046/pericytes-and-their-potential-in-regenerative-medicine-across-species
#17
REVIEW
C L Esteves, F X Donadeu
The discovery that pericytes are in vivo counterparts of Mesenchymal Stem/Stromal Cells (MSCs) has placed these perivascular cells in the research spotlight, bringing up hope for a well-characterized cell source for clinical applications, alternative to poorly defined, heterogeneous MSCs preparations currently in use. Native pericytes express typical MSC markers and, after isolation by fluorescence-activated cell sorting, display an MSC phenotype in culture. These features have been demonstrated in different species, including humans and horses, the main targets of regenerative treatments...
September 20, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28926198/serum-free-human-msc-medium-supports-consistency-in-human-but-not-in-equine-adipose-derived-multipotent-mesenchymal-stromal-cell-culture
#18
Susanna Schubert, Walter Brehm, Aline Hillmann, Janina Burk
For clinical applications of multipotent mesenchymal stromal cells (MSCs), serum-free culture is preferable to standardize cell products and prevent contamination with pathogens. In contrast to human MSCs, knowledge on serum-free culture of large animal MSCs is limited, despite its relevance for preclinical studies and development of veterinary cellular therapeutics. This study aimed to evaluate the suitability of a commercially available serum-free human MSC medium for culturing equine adipose-derived MSCs in comparison with human adipose MSCs...
September 19, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28914994/hessian-based-quantitative-image-analysis-of-host-pathogen-confrontation-assays
#19
Zoltan Cseresnyes, Kaswara Kraibooj, Marc Thilo Figge
Host-fungus interactions have gained a lot of interest in the past few decades, mainly due to an increasing number of fungal infections that are often associated with a high mortality rate in the absence of effective therapies. These interactions can be studied at the genetic level or at the functional level via imaging. Here, we introduce a new image processing method that quantifies the interaction between host cells and fungal invaders, for example, alveolar macrophages and the conidia of Aspergillus fumigatus...
September 15, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28906588/surface-glycan-pattern-of-canine-equine-and-ovine-bone-marrow-derived-mesenchymal-stem-cells
#20
Salvatore Desantis, Gianluca Accogli, Antonio Crovace, Edda G Francioso, Alberto Maria Crovace
The use of bone marrow-derived mesenchymal stem cells (MSCs) for clinical and experimental studies is increasing, but full characterization of MSCs in veterinary species is hindered by the variability in species-specific cell surface marker expression and antibody cross reactivity. Recent studies demonstrated that the glycans in the glycocalyx of MSCs are promising candidates as cell biomarkers. In the present study, we analyzed the glycocalyx of canine MSCs (cMSCs), ovine MSCs (oMSCs), and equine MSCs (eMSCs) using a cell microarray procedure in which MSCs were spotted on microarray slides and incubated with a panel of 14 biotinylated lectins and Cy3-conjugated streptavidin...
September 14, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
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