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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/30211979/vtx-1-liquid-biopsy-system-for-fully-automated-and-label-free-isolation-of-circulating-tumor-cells-with-automated-enumeration-by-bioview-platform
#1
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
https://www.readbyqxmd.com/read/30211978/nadh-autofluorescence-a-marker-on-its-way-to-boost-bioenergetic-research
#2
REVIEW
Patrick M Schaefer, Sviatlana Kalinina, Angelika Rueck, Christine A F von Arnim, Bjoern von Einem
More than 60 years ago, the idea was introduced that NADH autofluorescence could be used as a marker of cellular redox state and indirectly also of cellular energy metabolism. Fluorescence lifetime imaging microscopy of NADH autofluorescence offers a marker-free readout of the mitochondrial function of cells in their natural microenvironment and allows different pools of NADH to be distinguished within a cell. Despite its many advantages in terms of spatial resolution and in vivo applicability, this technique still requires improvement in order to be fully useful in bioenergetics research...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30211977/microfluidic-based-optical-microscopes-on-chip
#3
REVIEW
Petra Paiè, Rebeca Martínez Vázquez, Roberto Osellame, Francesca Bragheri, Andrea Bassi
Last decade's advancements in optofluidics allowed obtaining an ever increasing integration of different functionalities in lab on chip devices to culture, analyze, and manipulate single cells and entire biological specimens. Despite the importance of optical imaging for biological sample monitoring in microfluidics, imaging is traditionally achieved by placing microfluidics channels in standard bench-top optical microscopes. Recently, the development of either integrated optical elements or lensless imaging methods allowed optical imaging techniques to be implemented in lab on chip systems, thus increasing their automation, compactness, and portability...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30211975/object-oriented-segmentation-of-cell-nuclei-in-fluorescence-microscopy-images
#4
Can Fahrettin Koyuncu, Rengul Cetin-Atalay, Cigdem Gunduz-Demir
Cell nucleus segmentation remains an open and challenging problem especially to segment nuclei in cell clumps. Splitting a cell clump would be straightforward if the gradients of boundary pixels in-between the nuclei were always higher than the others. However, imperfections may exist: inhomogeneities of pixel intensities in a nucleus may cause to define spurious boundaries whereas insufficient pixel intensity differences at the border of overlapping nuclei may cause to miss some true boundary pixels. In contrast, these imperfections are typically observed at the pixel-level, causing local changes in pixel values without changing the semantics on a large scale...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30211969/in-situ-detection-of-cd73-cd90-cd105-lineage-mesenchymal-stromal-cells-in-human-placenta-and-bone-marrow-specimens-by-chipcytometry
#5
Christine Consentius, Anja Mirenska, Anke Jurisch, Simon Reinke, Markus Scharm, Ana Claudia Zenclussen, Christian Hennig, Hans-Dieter Volk
Mesenchymal stromal cells (MSCs) support endogenous regeneration and present therefore promising opportunities for in situ tissue engineering. They can be isolated and expanded from various tissues, for example, bone marrow, adipose tissue, or placenta. The minimal consensus definition criteria of ex vivo expanded MSCs requires them to be positive for CD73, CD90, and CD105 expression, while being negative for CD34, CD45, CD14, CD19, and HLA-DR. This study aimed to compare the in situ phenotype of MSCs with that of their culture-expanded progeny...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30211968/evaluation-of-bone-marrow-microenvironment-could-change-how-myelodysplastic-syndromes-are-diagnosed-and-treated
#6
REVIEW
Carmen Mariana Aanei, Lydia Campos Catafal
Myelodysplastic syndromes are a heterogeneous group of clonal hematopoietic disorders. However, the therapies used against the hematopoietic stem cells clones have limited efficacy; they slow the evolution toward acute myeloid leukemia rather than stop clonal evolution and eradicate the disease. The progress made in recent years regarding the role of the bone marrow microenvironment in disease evolution may contribute to progress in this area. This review presents the recent updates on the role of the bone marrow microenvironment in myelodysplastic syndromes pathogenesis and tries to find answers regarding how this information could improve myelodysplastic syndromes diagnosis and therapy...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30211967/proteomic-profiling-of-native-unpassaged-and-culture-expanded-mesenchymal-stromal-cells-msc
#7
Erika Moravcikova, E Michael Meyer, Mirko Corselli, Vera S Donnenberg, Albert D Donnenberg
Human culture-expanded mesenchymal stromal cells (MSC) are being considered for multiple therapeutic applications because of their regenerative and anti-inflammatory properties. Although a large number of MSC can be propagated from a small initial sample, several lines of evidence indicate that MSC lose their immunosuppressive and regenerative potency aftaer multiple passages. In this report, we use the FACSCAP Lyoplate proteomic analysis system to detect changes in cell surface protein expression of CD45- /CD31- /CD34- /CD73+ /CD105+ stromal cells in unpassaged bone marrow (BM) and through 10 serial culture passages...
September 13, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30184320/automated-rare-single-cell-picking-with-the-als-cellcelector%C3%A2
#8
Constantin Nelep, Jens Eberhardt
Molecular analysis of rare single cells like circulating tumor cells (CTCs) from whole blood patient samples bears multiple challenges. One of those challenges is the efficient and ideally loss-free isolation of CTCs over contaminating white and red blood cells. While there is a multitude of commercial and non-commercial systems available for the enrichment of CTCs their cell output does not deliver the purity most molecular analysis methods require. Here we describe the ALS CellCelector™ which can solve this challenge allowing the retrieval of 100% pure single CTCs from blood processed by different upstream enrichment techniques...
September 5, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30176186/determination-of-the-proliferative-fractions-in-differentiating-hematopoietic-cell-lineages-of-normal-bone-marrow
#9
Kelly P H Nies, Raisa Kraaijvanger, Kim H K Lindelauf, Roosmarie J M R Drent, Ronald M J Rutten, Frans C S Ramaekers, Math P G Leers
Because of the proven prognostic value of Ki-67 as a proliferation marker in several types of solid cancers, our goal is to develop and validate a multiparameter flow cytometric assay for the determination of the Ki-67 expression in hemato-oncological diseases. The aim of the present study was to establish the reference values for the fraction of Ki-67 positive cells in and during maturation of individual hematopoietic cell lineages present in normal bone marrow. Aspirates derived from femoral heads of 50 patients undergoing a hip replacement were used for the flow cytometric quantification of Ki-67 expression in the different hematopoietic cell populations of healthy bone marrow...
September 3, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30176185/automatically-generate-two-dimensional-gating-hierarchy-from-clustered-cytometry-data
#10
Xingyu Yang, Peng Qiu
Cytometry is an important technique widely used in medicine and biological research. Biologists traditionally analyze single-cell cytometry data by manual gating, which can be subjective and labor intensive. To address this issue, many automated and semiautomated methods have been developed. These advanced methods are designed to speed up and standardize the analysis of cytometry data, but their popularity is limited by their visualizations which are not intuitive to biologists who are accustomed to the conventional biaxial gating plots...
September 3, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30176184/spectral-imaging-of-fret-based-sensors-reveals-sustained-camp-gradients-in-three-spatial-dimensions
#11
Naga S Annamdevula, Rachel Sweat, John R Griswold, Kenny Trinh, Chase Hoffman, Savannah West, Joshua Deal, Andrea L Britain, Kees Jalink, Thomas C Rich, Silas J Leavesley
Cyclic AMP is a ubiquitous second messenger that orchestrates a variety of cellular functions over different timescales. The mechanisms underlying specificity within this signaling pathway are still not well understood. Several lines of evidence suggest the existence of spatial cAMP gradients within cells, and that compartmentalization underlies specificity within the cAMP signaling pathway. However, to date, no studies have visualized cAMP gradients in three spatial dimensions (3D: x, y, z).This is in part due to the limitations of FRET-based cAMP sensors, specifically the low signal-to-noise ratio intrinsic to all intracellular FRET probes...
September 3, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30157315/random-colocalization-and-split-of-alk-break-apart-probe-signals-potential-concerns-in-the-real-life-diagnosis-of-non-small-cell-lung-cancer
#12
EDITORIAL
Arnaud Uguen
No abstract text is available yet for this article.
August 29, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30110138/flowio-flow-cytometry-standard-conformance-testing-editing-and-export-tool
#13
Miroslav Koblížek, Anastasia Lebedeva, Karel Fišer
The Flow Cytometry Standard (FCS) format is a widely accepted norm for storing Flow Cytometry (FCM) data. Its goal as a standard is to allow FCM data sharing and re-analysis. Over more than three decades of its existence FCS has evolved into a well-defined, flexible file format reflecting technical changes in the FCM field. Its flexibility as well as rising numbers of instrument vendors leads to suboptimal implementations of FCS in some cases. Such situations compromise the primary goal of the standard and hinder the ability to reproduce FCM analyses...
August 15, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30107098/comparison-of-jc-1-and-mitotracker-probes-for-mitochondrial-viability-assessment-in-stored-canine-platelet-concentrates-a-flow-cytometry-study
#14
Natália Aydos Marcondes, Silvia Resende Terra, Camila Serina Lasta, Nicole Regina Capacchi Hlavac, Magnus Larruscaim Dalmolin, Luciana de Almeida Lacerda, Gustavo Adolpho Moreira Faulhaber, Félix Hilário Díaz González
Mitochondria perform crucial roles in many biochemical processes, and mitochondrial depolarization is an early sign of platelet apoptosis. The mitochondrial membrane potential is usually evaluated through JC-1 probe, but it can also be assessed with MitoTracker probes. Our aim was to evaluate mitochondrial viability in stored canine platelet concentrates (PCs) with the fluorescent probes JC-1 and MitoTracker. Platelets from 22 canine PCs were stained with JC-1 and MitoTracker probes on days 1, 3, and 5 of storage...
August 14, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30107096/multiplexed-fluorescence-microscopy-reveals-heterogeneity-among-stromal-cells-in-mouse-bone-marrow-sections
#15
Karolin Holzwarth, Ralf Köhler, Lars Philipsen, Koji Tokoyoda, Valeriia Ladyhina, Carolina Wählby, Raluca A Niesner, Anja E Hauser
The bone marrow (BM) consists of multiple, structured micro-environmental entities-the so called niches, which contain hematopoietic cells as well as stromal cells. These niches fulfill a variety of functions, such as control of the hematopoietic stem cell pool, differentiation of hematopoietic cells, and maintenance of immunological memory. However, due to the molecular and cellular complexity and a lack of suitable histological multiplexing methods, the composition of the various BM niches is still elusive...
August 14, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30107082/the-parsortix%C3%A2-cell-separation-system-a-versatile-liquid-biopsy-platform
#16
M Craig Miller, Peggy S Robinson, Christopher Wagner, Daniel J O'Shannessy
Cancer cells from solid tumors can enter the circulatory system and survive to subsequently form distant metastases. The CellSearch® system (Menarini-Silicon Biosystems, Huntingdon Valley, PA) was the first, FDA-cleared system that provided a reliable tool for the investigation of circulating tumor cells (CTCs), which have been shown to be strongly associated with poor survival and therapy failure. Since that time, a number of new technologies have been introduced to improve CTC detection and/or isolation for further characterization...
August 14, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30107080/flow-cytometric-detection-of-most-proteins-in-the-cell-surface-proteome-is-unaffected-by-trypsin-treatment
#17
Vera S Donnenberg, Mirko Corselli, Daniel P Normolle, E Michael Meyer, Albert D Donnenberg
Flow cytometry is often performed on adherent cells or solid tissues that have been released from their growth substrate or disaggregated by enzymatic digestion. Although detection of strongly expressed cell surface proteins following such procedures indicates that many survive treatment with proteolytic enzymes, applications such as cell surface proteomics involve assessment of the expression of more than 200 proteins and it is important to know how to interpret negative results. To address this problem, we performed flow cytometry-based cell surface proteomic analysis on two non-adherent cell lines, THP1 and K562, after mock and authentic trypsin treatment, according to a widely used protocol to remove adherent cells (0...
August 14, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30089197/quantitative-chromogenic-immunohistochemical-image-analysis-in-cellprofiler-software
#18
REVIEW
V Tollemar, N Tudzarovski, E Boberg, A Törnqvist Andrén, A Al-Adili, K Le Blanc, K Garming Legert, M Bottai, G Warfvinge, R V Sugars
Visual grading of chromogenically stained immunohistochemical (IHC) samples is subjective, time consuming, and predisposed to considerable inter- and intra-observer variations. The open-source digital analysis software, CellProfiler has been extensively used for fluorescently stained cells/tissues; however, chromogenic IHC staining is routinely used in both pathological and research diagnostics. The current investigation aimed to compare CellProfiler quantitative chromogenic IHC analyses against the gold standard manual counting...
August 8, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30080307/clearcell%C3%A2-fx-a-label-free-microfluidics-technology-for-enrichment-of-viable-circulating-tumor-cells
#19
Yifang Lee, Guofeng Guan, Ali Asgar Bhagat
Circulating tumor cells (CTCs) dissociate from primary tumor into the bloodstream, and carry with them cancer's fingerprints as well as the potential to turn aggressive and metastasize. In order to understand CTCs and develop clinical utility, different methods of enrichment and isolation of CTCs can be used. Here, we report the use of a label-free platform, ClearCell® FX which isolates CTCs by their mechanical features and its advantages. The technology utilizes Dean Flow Fractionation (DFF) principle in a spiral microfluidics system to separate the larger CTCs from smaller blood cells...
August 6, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/30071132/a-rapid-single-cell-sorting-verification-method-using-plate-based-image-cytometry
#20
Eric S Zigon, Suzanne M Purseglove, Vasilis Toxavidis, William Rice, John Tigges, Leo Li-Ying Chan
Single cell sorting is commonly used for ensuring monoclonality and producing homogenous target cell populations. Current single cell verification methods involve manually confirming the existence of single cells or colonies in a well using a standard light microscope. However, the manual verification method is time-consuming and highly tedious, which prompts a need for an accurate and rapid detection method for verifying single cell sorting capability. Here, we demonstrate a rapid single cell sorting verification method using the Celigo Image Cytometer...
August 2, 2018: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
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