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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/30451364/a-flow-cytometric-study-of-er-stress-and-autophagy
#1
A Popat, A A Patel, G Warnes
The mechanistic link between ER stress, autophagy, and resultant cell death was investigated by the use of drugs Thapsigargin (Tg) and Chloroquine (CQ) with prior induction and or blockade of autophagy and apoptosis which modulated the ER stress response and resultant form of cell death. All these biological processes can be measured flow cytometrically allowing the determination of the type of cell death, G1 cell cycle arrest, cell cycle dependent measurement of ER stress transducer PERK, misfolded proteins, reticulophagy, and autophagy marker LC3B...
November 19, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30450827/fast-and-quantitative-evaluation-of-human-leukocyte-interaction-with-aspergillus-fumigatus-conidia-by-flow-cytometry
#2
Susann Hartung, Christopher Rauh, Thi Ngoc Mai Hoang, Susanne Jahreis, Kathleen Wagner, Juliane Macheleidt, Axel A Brakhage, Silke Rummler, Andreas Hochhaus, Marie von Lilienfeld-Toal
Systemic infections with the opportunistic mold Aspergillus fumigatus are a great threat to immunocompromised patients such as transplant recipients. Immunological research on A. fumigatus involves the measurement of phagocytosis of fungal conidia (spores) by human phagocytes. Here, we present a fast and flexible way to analyze phagocytosis by flow cytometry using fluorescein isothiocyanate (FITC) labeling of conidia prior to co-incubation with human leukocytes and an anti-FITC counterstaining step postincubation to allow the discrimination of internalized and adherent conidia...
November 19, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30427580/vycap-s-puncher-technology-for-single-cell-identification-isolation-and-analysis
#3
Michiel Stevens, Lisa Oomens, Joska Broekmaat, Joris Weersink, Fikri Abali, Joost Swennenhuis, Arjan Tibbe
Here we present the Puncher technology for the isolation of single cells. This technology combines a silicon chip with microwells, fluorescence imaging, and a punching method to isolate and transfer the single cells to standard reaction tubes. The technology is compatible with commercially available downstream workflows and instrumentation. Here we focus on the isolation of CTC but the Puncher technology can be applied to isolate single cells from liquid biopsies and more general from cell suspensions. It is especially suited for cell suspensions that contain: Cells of interest at a frequency of 1 per 10,000 or less A low total number of cells ranging from 1 to 100,000, that are present in a volume of 0...
November 14, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30423208/deep-ultraviolet-lasers-for-flow-cytometry
#4
William Telford, Thierry Georges, Clint Miller, Pascal Voluer
Modern flow cytometers require multiple laser wavelengths to excite the wide variety of fluorescent probes now available for high-dimensional analysis. Ultraviolet (UV) lasers (typically solid state 355 nm) have become a critical excitation source for the Brilliant Ultraviolet (BUV) series of polymer fluorochromes. The BUV dyes have pushed the number of fluorescent probes available for simultaneous analysis to nearly 30, allowing an unprecedented level of precision for immune cell analysis. However, immunologists are already seeking analyze more than 30 simultaneous parameters, requiring both new fluorochromes and corresponding laser wavelengths...
November 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30422397/concurrent-detection-of-lysosome-and-tissue-transglutaminase-activation-in-relation-to-cell-cycle-position-during-apoptosis-induced-by-different-anticancer-drugs
#5
H Dorota Halicka, Jiangwei Li, Hong Zhao, Zbigniew Darzynkiewicz
Described is the new cytometric approach do detect either stimulation or a collapse of lysosomal proton pump (lysosomes rupture) combined with activation of transglutaminase 2 (TG2) during induction of apoptosis. Apoptosis of human lymphoblastoid TK6 cells was induced by combination of 2-deoxyglucose with the isoquinoline alkaloid berberine, by DNA topoisomerase I inhibitor camptothecin, its analog topotecan, topoisomerase II inhibitors etoposide or mitoxantrone, as well as by the cytotoxic anticancer ribonuclease ranpirnase (onconase)...
November 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30382613/could-the-key-be-hidden-in-the-lysosomes
#6
Gloria Juan
No abstract text is available yet for this article.
November 1, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30382608/cd40-signaling-germinal-center-b-cells-go-at-their-own-pace
#7
Thomas Liechti
No abstract text is available yet for this article.
November 1, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30378734/a-comparative-study-of-two-separation-methods-to-isolate-monocytes
#8
Ronald Weiss, Wilhelm Gerdes, Franziska Leonhardt, Rommy Berthold, Ulrich Sack, Anja Grahnert
Separation of specific blood cells is necessary for a deeper insight into their role in health and disease. To obtain such cells, efficient and robust isolation methods are needed. We compare here the Fab-based Traceless Affinity Cell Selection (TACS®) technology and the Magnetic Activated Cell Sorting (MACS®) technology to isolate human monocytes from whole blood and buffy coats as well as the differentiation of the isolated monocytes to dendritic cells (DCs). TACS® is a positive selection technology using immune affinity chromatography based on CD-specific low affinity Fab-fragments for the reversible capture and release of target cells...
October 31, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30343512/two-photon-fluorescence-lifetime-imaging-of-intrinsic-nadh-in-three-dimensional-tumor-models
#9
Anh Cong, Rafaela M L Pimenta, Hong Bok Lee, Venkatram Mereddy, Jon Holy, Ahmed A Heikal
Most studies using intrinsic NAD(P)H as biomarkers for energy metabolism and mitochondrial anomalies have been conducted in routine two-dimensional (2D) cell culture formats. Cellular metabolism and cell behavior, however, can be significantly different in 2D cultures from that in vivo. As a result, there are emerging interests in integrating noninvasive, quantitative imaging techniques of NAD(P)H with in vivo-like three-dimensional (3D) models. The overall features and metabolic responses of the murine breast cancer cells line 4T1 in 2D cultures were compared with those in 3D collagen matrix using integrated optical micro-spectroscopy...
October 21, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30369063/fluorescence-lifetime-shifts-of-nad-p-h-during-apoptosis-measured-by-time-resolved-flow-cytometry
#10
Faisal Alturkistany, Kapil Nichani, Kevin D Houston, Jessica P Houston
Autofluorescence from the intracellular metabolite, NAD(P)H, is a biomarker that is widely used and known to reliably screen and report metabolic activity as well as metabolic fluctuations within cells. As a ubiquitous endogenous fluorophore, NAD(P)H has a unique rate of fluorescence decay that is altered when bound to coenzymes. In this work we measure the shift in the fluorescence decay, or average fluorescence lifetime (1-3 ns), of NAD(P)H and correlate this shift to changes in metabolism that cells undergo during apoptosis...
October 19, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30369050/blood-based-biopsies-clinical-utility-beyond-circulating-tumor-cells
#11
Cha-Mei Tang, Peixuan Zhu, Shuhong Li, Olga V Makarova, Platte T Amstutz, Daniel L Adams
Circulating tumor cells (CTCs), epithelial-mesenchymal transition (EMT) cells, as well as a number of circulating cancer stromal cells (CStCs) are known to shed into the blood of cancer patients. Individually, and together, these cells provide biological and clinical information about the cancers. Filtration is a method used to isolate all of these cells, while eliminating red and white blood cells from whole peripheral blood. We have previously shown that accurate identification of these cell types is paramount to proper clinical assessment by describing the overlapping phenotypes of CTCs to one such CStC, the cancer-associated macrophage-like cell (CAML)...
October 19, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30329217/in-vivo-metabolic-and-shg-imaging-for-monitoring-of-tumor-response-to-chemotherapy
#12
Maria M Lukina, Varvara V Dudenkova, Lyubov' E Shimolina, Ludmila B Snopova, Elena V Zagaynova, Marina V Shirmanova
Although chemotherapy remains one of the main types of treatment for cancer, treatment failure is a frequent occurrence, emphasizing the need for new approaches to the early assessment of tumor response. The aim of this study was to search for indicators based on optical imaging of cellular metabolism and of collagen in tumors in vivo that enable evaluation of their response to chemotherapy. The study was performed on a mouse colorectal cancer model with the use of cisplatin, paclitaxel, and irinotecan. The metabolic activity of the tumor cells was assessed using fluorescence lifetime imaging of the metabolic cofactor reduced nicotinamide adenine dinucleotide (phosphate), NAD(P)H...
October 17, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30296355/autofluorescence-lifetime-imaging-of-cellular-metabolism-sensitivity-toward-cell-density-ph-intracellular-and-intercellular-heterogeneity
#13
Jenu V Chacko, Kevin W Eliceiri
Autofluorescence imaging (AFI) has greatly accelerated in the last decade, way past its origins in detecting endogenous signals in biological tissues to identify differences between samples. There are many endogenous fluorescence sources of contrast but the most robust and widely utilized have been those associated with metabolism. The intrinsically fluorescent metabolic cofactors nicotinamide adenine dinucleotide (NAD+ /NADH) and flavin adenine dinucleotide (FAD/FADH2 ) have been utilized in a number of AFI applications including basic research, clinical, and pharmaceutical studies...
October 8, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30277662/combining-fluorescent-cell-barcoding-and-flow-cytometry-based-phospho-erk1-2-detection-at-short-time-scales-in-adherent-cells
#14
Sonal Manohar, Prachi Shah, Sharmila Biswas, Anam Mukadam, Madhura Joshi, Ganesh Viswanathan
Detection of levels of intracellular phospho-proteins is key to analyzing the dynamics of signal transduction in cellular systems. Cell-to-cell variability in the form of differences in protein level in each cell affects signaling and is implicated in prognosis of many diseases. Quantitative analysis of such variability necessitate measuring the protein levels at single-cell resolution. Single-cell intracellular protein abundance detection in statistically significant number of adherent cells for short time sampling points post stimulation using classical flow cytometry (FCM) technique has thus far been a challenge due to the detrimental effects of cell detachment methods on the cellular machinery...
October 2, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30277660/the-rarecyte%C3%A2-platform-for-next-generation-analysis-of-circulating-tumor-cells
#15
Eric P Kaldjian, Arturo B Ramirez, Yao Sun, Daniel E Campton, Jeffrey L Werbin, Paulina Varshavskaya, Steven Quarre, Tad George, Anup Madan, C Anthony Blau, Ronald Seubert
Circulating tumor cells (CTCs) can reliably be identified in cancer patients and are associated with clinical outcome. Next-generation "liquid biopsy" technologies will expand CTC diagnostic investigation to include phenotypic characterization and single-cell molecular analysis. We describe here a rare cell analysis platform designed to comprehensively collect and identify CTCs, enable multi-parameter assessment of individual CTCs, and retrieve single cells for molecular analysis. The platform has the following four integrated components: 1) density-based separation of the CTC-containing blood fraction and sample deposition onto microscope slides; 2) automated multiparameter fluorescence staining; 3) image scanning, analysis, and review; and 4) mechanical CTC retrieval...
October 2, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30277658/the-anatomy-of-single-cell-mass-cytometry-data
#16
REVIEW
Lars R Olsen, Michael D Leipold, Christina B Pedersen, Holden Terry Maecker
Mass cytometry enables the measurement of up to 50 features on single cell. This has catalyzed a shift toward multidimensional data analysis methods, rather than the manual gating strategies as traditionally for in flow cytometry data. This shift means that data scientists are involved in the analysis process to an increasing degree. As the data is analyzed in a more unbiased fashion, where noisy or uninformative observations are not easily excluded, a deeper knowledge of the origin, noise, and modalities of the data is therefore needed to embark on useful data analysis...
October 2, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30246927/how-to-agree-on-a-ctc-evaluating-the-consensus-in-circulating-tumor-cell-scoring
#17
Leonie L Zeune, Sanne de Wit, A M Sofie Berghuis, Maarten J IJzerman, Leon W M M Terstappen, Christoph Brune
For using counts of circulating tumor cells (CTCs) in the clinic to aid a physician's decision, its reported values will need to be accurate and comparable between institutions. Many technologies have become available to enumerate and characterize CTCs, thereby showing a large range of reported values. Here we introduce an Open Source CTC scoring tool to enable comparison of different reviewers and facilitate the reach of a consensus on assigning objects as CTCs. One hundred images generated from two different platforms were used to assess concordance between 15 reviewers and an expert panel...
September 24, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30240134/morpho-functional-characteristics-of-bone-marrow-multipotent-mesenchymal-stromal-cells-after-activation-or-inhibition-of-epidermal-growth-factor-and-toll-like-receptors-or-treatment-with-dna-intercalator-cisplatin
#18
Iuliia Golovynska, Olesia Kalmukova, Hanna M Svitina, Vitaliy M Kyryk, Volodimir A Shablii, Nataliya V Senchylo, Galyna V Ostrovska, Mykola Dzerzhinskyi, Yurii V Stepanov, Sergii Golovynskyi, Tymish Y Ohulchanskyy, Liwei Liu, Liudmila V Garmanchuk, Junle Qu
This study is aimed to reveal morphological and functional changes in multipotent mesenchymal stromal cells (MSCs) isolated from the rat bone marrow after: (i) activation of Toll-like receptors (TLRs) with teichoic acid (TA), (ii) impact on epidermal growth factor (EGF) receptors with activator EGF or inhibitor Herceptin, and (iii) treatment with DNA intercalator Cisplatin. According to our results, TA and EGF cause an increase in the synthesis of glycosaminoglycans, c-Myc content, and protein in the MSC cytoplasm...
September 21, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30240113/spectrally-and-spatially-resolved-laser-induced-photobleaching-of-endogenous-flavin-fluorescence-in-cardiac-myocytes
#19
Alzbeta Marcek Chorvatova, Jana Kirchnerova, Michal Cagalinec, Anton Mateasik, Dusan Chorvat
Naturally occurring endogenous fluorescence of flavins, arising in response to excitation by visible light, offers broad opportunity to investigate mitochondrial metabolic state directly in living cells and tissues, including in clinical settings. However, photobleaching, the loss of the autofluorescence intensity following prolonged exposure to light is an inherent phenomenon occurring during the fluorescence acquisition, which can have a negative impact on the recorded data, particularly in the context of measurement of metabolic modulations in pathophysiological conditions...
September 21, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30211979/vtx-1-liquid-biopsy-system-for-fully-automated-and-label-free-isolation-of-circulating-tumor-cells-with-automated-enumeration-by-bioview-platform
#20
Elodie Sollier-Christen, Corinne Renier, Tal Kaplan, Elad Kfir, Steve C Crouse
Clinicians continue to rely on invasive tissue biopsies as a mean to assess a patient's disease and prescribe appropriate treatment regimens. Biopsies not only are risky and expensive but also limit the understanding of disease. Circulating tumor cells (CTCs) can be isolated from a simple blood draw and offer a promising potential to both diagnose and monitor cancer progression. The VTX-1 Liquid Biopsy System automates the isolation of clinically relevant CTC populations, while simplifying their collection for easy analysis, ultimately expanding the clinical possibilities for CTCs...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
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