Computational tools will be employed for the analysis of multiparametric flow cytometry data. Classical two-dimensional scatter plots analysis cannot be sufficient for the multidimensional nature of the flow cytometric data, especially when many parameters are simultaneously combined for studying the phenotype, effector function and the polyfunctionality of activated immune cells. Computational tools will be employed to analyze, visualize and interpret large amounts of cell data in an automated and unbiased way. Multiparametric flow cytometry can be particularly suitable for deep analysis of both T and B responses after vaccination, allowing to measure the frequency, the phenotype and the functional features of antigen-specific cells. The automated analysis allows to identify, in an unbiased way, all the cellular phenotypes elicited by vaccination, including not only terminally differentiated cells but also transient stages of differentiation, thus providing important information on activation ...
TY - JOUR. T1 - Flow cytometric detection and quantitative analysis of the GM-CSF receptor in human granulocytes and comparison with the radioligand binding assay. AU - Stacchini, Alessandra. AU - Fubini, Lidia. AU - Aglietta, Massimo. PY - 1996/8/1. Y1 - 1996/8/1. N2 - A flow cytometric method to quantify the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFr) on human cells is described. The number of GM-CSFr binding sites on human neutrophils was assessed by using different bead standards. Results were compared with those from conventional receptor quantification, which was performed by using the radioligand binding assay. A high degree of correlation was found between the two methods, although quantitative evaluation of GM-CSFr expression on neutrophils performed by flow cytometry revealed a somewhat higher number of receptor molecules per cell than with that determined by Scatchard analysis. By the flow quantitative approach, we measured the GM-CSFr on mobilized ...
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Traditional flow cytometry methods have been remarkably successful at detecting and sorting specific cells in a sample consisting of many cell types. However, a significant limitation of these methods is their inability to be applied in-vivo. Our research focuses on creating and perfecting novel flow cytometry methods which resolve this problem. Previous work includes the development of a two-photon in-vivo flow cytometry system. The use of multiphoton excitation creates an extremely narrow excitation region, making it possible to record signals from single cells as they propagate through the bloodstream. This eliminates the complicated fluid dynamics required for conventional flow cytometry, and also allows the user to focus on a single blood vessel in a living animal. Current and future work includes investigations of coherent control and the use of fiber optics for monitoring in the bloodstream itself.. ...
We report the development of a novel flow cytometry-based Ig capture assay (ICA) for the identification and sorting of individual Ab-secreting cells based on their Ag reactivity. The ICA represents a fast and versatile tool for single-cell sorting of peripheral plasmablasts, streamlining subsequent Ab analysis, and cloning. We demonstrate the utility of the assay by isolating Ag-reactive plasmablasts from cryopreserved PBMC obtained from volunteers vaccinated with a recombinant HIV envelope protein. To show the specificity of the ICA, we produced Ag-specific Abs from these cells and subsequently verified their Ag reactivity via ELISA. Furthermore, we used the ICA to track Ag-specific plasmablast responses in HIV-vaccine recipients over a period of 42 d and performed a head-to-head comparison with a conventional B cell ELISpot. Results were highly comparable, highlighting that this assay is a viable alternative for monitoring Ag-specific plasmablast responses at early time points after infection ...
Commercial organizations. Regional Segments:. Regional segmentation details the regional aspects of the global Flow Cytometry market. and explains the restrictive framework thats probably to impact the market. It highlights the political scenario in the market and the anticipates its influence on the global Flow Cytometry market.. • Middle East and Africa (Egypt and GCC Countries). • North America (United States, Mexico, and Canada). • South America (Brazil etc.). • Europe (Germany, Turkey, Russia UK, Italy, France, etc.). Get UPTO 25% OFF On Flow Cytometry Market Report (Valid Till 15Jan 2020). Inquire/Speak To Expert for Further Detailed Information About Flow Cytometry Report: https://marketresearch.biz/report/flow-cytometry-market/#inquiry. Table of Contents:. Chapter One: Global Flow Cytometry Market Overview. 1.1 Flow Cytometry Preface. Chapter Two: Global Flow Cytometry Market Analysis. 2.1 Flow Cytometry Report Description. 2.1.1 Flow Cytometry Market Definition and Scope. 2.2 ...
Flow cytometry is conventionally used to measure cell-surface antigen expression. However, many antigens are found within the cytoplasm, and it is necessary to fix and permeabilize cells to enable antibodies to gain access to them. In this study we have established the conditions for studying intracellular antigens in human trophoblast cells by flow cytometry using an antibody to TAP1 (a key molecule in the process of Class I MHC assembly). We have previously shown by immunocytochemistry that TAP1 expression is apparently greater on Class 1 positive extravillous cytotrophoblast than on any other fetal or maternal tissue. However, as immunohistochemistry is not quantitative we have used three-colour flow cytometry to measure the expression of TAP1 in different trophoblast populations. Villous and extravillous cytotrophoblast were identified in first trimester and term placental and decidual digests on the basis of their expression of cytokeratin and Class I MHC antigens. The level of expression of TAP1
Custom flow cytometry on clinical specimens is an eagle-i resource of type Material processing service at The University of Pennsylvania.
Hi Philipp, You can get devel versions of R at ftp://ftp.stat.math.ethz.ch/Software/R/. I believe the idea has always been that BioC devel (i.e., 2.6) works with R devel; iFlow is in BioC 2.6 so that should work. However, iFlow may not necessarily need the latest R. I havent tried the latest version but I have been playing with iFlow back in 2009 using R 2.9 or so. You can install iFlow from sources from the SVN repository (https://hedgehog.fhcrc.org/bioconductor/trunk/madman/Rpacks) user readonly, password readonly. To install it, run the R CMD INSTALL ,path to your iFlow,. The only pain with this approach is that you will have to manually resolve all the dependencies. Good luck. Cheers, Josef , Date: Mon, 08 Feb 2010 11:52:41 +0100 , From: Philipp Meng ,philipp.meng at meduniwien.ac.at, , To: Martin Morgan ,mtmorgan at fhcrc.org, , Cc: bioconductor ,bioconductor at stat.math.ethz.ch, , Subject: Re: [BioC] Installing problems of Flow Cytometry on Ubuntu , Karmic , Message-ID: , ...
Influenza remains a significant global public health burden, despite substantial annual vaccination efforts against circulating virus strains. As a re
Some snippets from your converted document: Designing and Implementing a High-Level Multicolor Flow Cytometry Assay Brent Wood MD PhD Department of Laboratory Medicine University of Washington Define Purpose of Assay • Most important question - What information is required? - What information is most important? • Prioritize • Compromises are inevitable - Simplest assay is best Outline • Instrument optimization • Compensation • Reagent selection and optimization • Panel validation • Data analysis Detector Optimization Detector Optimization PMT Voltage Green = 400 V, Blue = 450 V, Red = 500 V, Violet = 550 V Detector Voltage 475 volts 575 volts 675 volts 775 volts 875 volts • Too low reduces sensitivity • Too high pushes bright signals off scale - Dilute with unlabeled antibody • May need to compromise Optimization • Daily optimization not practical • Once optimized, do NOT change instrument settings unless change in performance - New optical filters - New PMT ...
Background: The neutral comet assay was devised to measure double-stranded DNA breaks, but it has also been used to measure apoptosis based on its characteristic DNA fragmentation patterns. There is still uncertainty about the reliability of this method. By comparing the comet assay with a flow cytometry method that uses Annexin V binding to apoptotic cells, we have provided further evidence for evaluating the usefulness of the comet assay for detecting apoptosis. Methods: Apoptosis was induced in human peripheral blood mononuclear cells (PBMC) by ionizing radiation and measured using the comet assay and a flow cytometry method that measures Annexin V and propidium iodide (PI) staining. Results: The Annexin V flow cytometry assay distinguished among early apoptosis, late apoptosis, and an apoptotic or necrotic phase in which the cells were labeled with both Annexin V and PI. The comet assay detected only the latter two phases of apoptosis. Conclusions: The comet assay is a useful tool for ...
TY - JOUR. T1 - Polychromatic flow cytometry analysis of CD34+ hematopoietic stem cells in cryopreserved early preterm human cord blood samples. AU - D'Alessio, F.. AU - Mirabelli, P.. AU - Gorrese, M.. AU - Scalia, G.. AU - Gemei, M.. AU - Mariotti, E.. AU - Di Noto, R.. AU - Martinelli, P.. AU - Fortunato, G.. AU - Paladini, D.. AU - Del Vecchio, L.. PY - 2011/1. Y1 - 2011/1. N2 - During the last decades, extended characterizations were performed of human full-term cord blood (hTCB) cells, but little information is available on human early preterm cord blood (hEPCB) hematopoietic stem cells (HSCs). In our study, we analyzed by flow cytometry 19 hEPCB and 17 hTCB samples. First, we observed that the percentage of CD34 PosCD45 Dim cells was higher in hEPCB compared with hTCB and that it decreased during 16th-20th week of pregnancy. Within the CD34 PosCD45 Dim population, we examined the expression of CD29, CD31, CD38, CD90, CD117, CD133, CD135, CD200, CD243, and CD338. We found that CD135 ...
Aims: To investigate the antibacterial efficacy of vancomycin towards Staphylococcus aureus under aerobic and anaerobic conditions, and to assess the influence of oxygen on the duration of the post-antibiotic effect (PAE) after exposure to vancomycin. Methods and Results: Culture-based techniques and flow cytometric measurements of 5-cyano-2,3-ditolyl tetrazolium chloride (an indicator of redox activity) and the membrane potential-sensitive fluorophore Sytox Green, were used to test four staphylococcal strains. The MICs for all strains, and the duration of PAE, were similar whether tested with or without oxygen. However, a fivefold logarithmic reduction in cell counts was observed in 10-15 h aerobically, depending on strain, compared with longer than 60 h in an anaerobic environment. Flow cytometric data correlated well with counts of colony-forming units under both aerobic and anaerobic conditions. Conclusions: The death rate of Staph. aureus exposed to vancomycin was greater in the presence of ...
TY - JOUR. T1 - Cell cycle analysis using flow cytometry. AU - Gray, J. W.. AU - Dolbeare, F.. AU - Pallavicini, M. G.. AU - Beisker, W.. AU - Waldman, F.. PY - 1986/1/1. Y1 - 1986/1/1. N2 - This manuscript reviews the utility of flow cytometry for the study of cell proliferation. The applications of univariate DNA distribution analysis to cytokinetic studies of asynchronous and perturbed cell populations are discussed briefly. The newly developed technique for simultaneous flow cytometric measurement of cellular DNA content and amount of incorporated bromodeoxyuridine is discussed in more detail. The cytochemistry required for this analysis is reviewed as are its applications to: (a) determination of the fractions of cells in the G1-, S- and G2 + M phases of the cell cycle; (b) determination of the G1-, S- and G2 + M phase durations and dispersions and growth fraction for asynchronous cells; (c) detection of ara-C resistant cells present at low frequency in an otherwise sensitive population; ...
The CellROX Orange Flow Cytometry Assay Kit enables the flow cytometric detection of reactive oxygen species (ROS) in live cells. The kit includes the novel, fluorogenic CellROX Orange Reagent, as well as SYTOX Red Dead Cell Stain, N-acetyl cysteine (an antioxidant, for negative control), and tert-b
Tomas Kalina, Standardization of cytometry and antibody validation. ​. There is an agreement in the field that interlaboratory reproducibility of flow cytometry measurements as well as whole studies might be improved by a consensual use of a methodological approach. Typically, a consensus is made on the crucial markers needed in the immunostaining panel, sometimes on the particular fluorochrome conjugates and rarely on a complete set of methods for sample preparation. The term standardization is used to describe the complete set of methodical steps, while harmonization is used for partial agreement on the method. Standardization can provide a platform for improved reproducibility of cytometry results over prolonged periods of time, across different sites and across different instruments. The different standardized approaches can and in fact should serve as benchmarking reference tools for the development of future flow cytometry studies. The cornerstone of each flow cytometry experiment ...
A fundamental tenet of scientific research is that the published results of any study have to be open to independent validation or refutation. The Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) establishes criteria for recording and reporting information about the flow cytometry experiment overview, samples, instrumentation and data analysis. It promotes consistent annotation of clinical, biological and technical issues surrounding a flow cytometry experiment by specifying the requirements for data content and by providing a structured framework for capturing information ...
Author(s): Uitrakul S, Hutton C, Veal GJ, Jamieson D. Publication type: Article. Publication status: Published. Journal: European Journal of Clinical Investigation. Year: 2019. Volume: 49. Issue: 7. Print publication date: 27/06/2019. Online publication date: 31/03/2019. Acceptance date: 26/03/2019. ISSN (print): 0014-2972. ISSN (electronic): 1365-2362. Publisher: Wiley-Blackwell. URL: https://doi.org/10.1111/eci.13115. DOI: 10.1111/eci.13115. ...
The aim of this study was to compare DNA content in hepatocyte and erythrocyte nuclei of the European sunbleak, Leucaspius delineatus, in relation to nuclear and cell size by means of flow cytometry and fluorescence microscopy. The DNA standards, chicken and rainbow trout erythrocytes, were prepared in parallel with both cell types, with initial separation of liver cells in pepsin solution followed by cell filtering. Standards and investigated cells were stained with a mixture of propidium iodide, citric acid, and Nonidet P40 in the presence of RNAse, and fluorescence of at least 50,000 nuclei was analyzed by flow cytometry. Average cell size was determined by flow cytometry, using fresh cell suspension in relation to latex beads of known diameter. The size of nuclei was examined on the basis of digital micrographs obtained by fluorescence microscopy after nuclei staining with DAPI. The sunbleaks erythrocyte nuclei contain 2.25 ± 0.06 pg of DNA, whereas the hepatocyte nuclei contain 2.46 ± ...
Circulating tumor cells are cells that detach from the primary tumor site and migrate to the bone marrow or other tissues where they can initiate a metastatic site. Liquid biopsies are an emerging tool in the past decades that enables us to detect Circulating Tumor Cells in patients blood. Flow cytometry is a powerful tool used in liquid biopsy diagnostics. This aims to prove the sensitivity and specificity of a flow cytometric panel for the detection of CTCs in breast cancer patients using healthy individuals samples as controls. The study was blinded to the data analyzing researcher. Statistical analysis followed and results show 86.9% area under the curve which indicates that the particular method can be very promising for diagnosing breast cancer.
Patients with uncharacteristic inflammatory symptoms such as long-standing fatigue or pain, or a prolonged fever, constitute a diagnostic and therapeutic challenge. The aim of the present study was to determine if an extended immunophenotyping of lymphocytes and monocytes including activation markers can define disease-specific patterns, and thus provide valuable diagnostic information for these patients. Whole blood from patients with gram-negative bacteraemia, neuroborreliosis, tuberculosis, acute mononucleosis, influenza or a mixed connective tissue disorders, as diagnosed by routine culture and serology techniques was analysed for lymphocyte and monocyte cell surface markers using a no-wash, no-lyse protocol for multi-colour flow cytometry method. The immunophenotyping included the activation markers HLA-DR and CD40. Plasma levels of soluble TNF alpha receptors were analysed by ELISA. An informative pattern was obtained by combining two of the analysed parameters: (i), the fractions of HLA-DR
Antigen uptake dendritic cells key step induction antigen-specific T-cell responses flow cytometry-based protocol describes analysis cell soluble anti
Investigation of signal transduction mechanisms is important for development of therapy and understanding complex life systems. In this study, to establish a novel recognition method of signal transduction pathways, we investigated signal transducers using flow cytometry. @The flow cytometric measurement shows a mean phosphorylation level (mean of fluorescence intensity, MFI) and a deviation of the phosphorylation level (coefficient variation, CV) in a cluster of cells. As a model of signal pathways, Jurkat cells (T cell leukemia cell line) were stimulated with interleukin-21 or interferon- , and signal transducers and activators of transcription (STATs) and extracellular signal-regulated kinase (ERK) 1/2 were measured using flow cytometry. Furthermore, peripheral blood was stimulated, and then various signal transducers of the lymphocytes and neutrophils were analyzed with MFI and CV. @After the stimulation, the increase of STATs MFI induced a temporal change of CV. On the other hand, the ...
Flow cytometry is designed to measure physical and biochemical characteristics of cells and cell-like particles using fluorescence. Fundamentally, any single-particle suspension (within a defined size range) can pass through the flow cytometer. Beads, for better or worse, are a sine qua non for the flow cytometrist. From quality control,to standardization, to compensation, there is a bead for every job. They are important - critical, even - for flow cytometry.
Sysmex analysers use the fluorescence flow cytometry method in the reticulocyte channel to measure fragmented red blood cells (FRC% and FRC#) as research parameters.. A specific area below the RBC area in the RET scattergram is used for identification of fragmented red blood cells. Due to the absence of nucleic acids in red blood cells the intensity of the measured side fluorescence signals (SFL) is extremely low. In addition, the high-angle forward scatter (FSC) is lower than that of intact red blood cells.. ...
Spring 2017 Fluorescence Calibration study - John Nolan. This years study aims to demonstrate a consensus approach to calibration and reporting of EV fluorescence measurements. The study has three tasks:. 1) Calibrate instrument fluorescence response using commercial reagents,. 2) Measure a common set of EV reference materials using a diverse set of flow cytometry methods and instruments, and report the data in a uniform manner such that results can be compared across methods and instruments, and. 3) Evaluate a prototype set of EV-scale fluorescence intensity and antibody binding standards that may be more appropriate for calibration and standardization of EV measurements.. All participants will perform a common set of measurements and report their methods and results according to draft Minimum Information about an EV Flow Cytometry measurement (MI EV FlowCyt) guidelines. The methods, results and guidelines that result from the study will be posted here and reported in a manuscript to be ...
The phosphatidylinositol 3-kinase (PI3K) pathway plays an important role in multiple myeloma (MM), a blood cancer associated with uncontrolled proliferation of bone marrow plasma cells. This study aimed to develop a robust clinical pharmacodynamic (PD) assay to measure the on-target PD effects of the selective PI3K inhibitor GDC-0941 in MM patients. We conducted an in vitro drug wash-out study to evaluate the feasibility of biochemical approaches in measuring the phosphorylation of S6 ribosomal protein (S6), one of the commonly used PD markers for PI3K pathway inhibition. We then developed a 7-color phospho-specific flow cytometry assay, or phospho flow assay, to measure the phosphorylation state of intracellular S6 in bone marrow aspirate (BMA) and peripheral blood (PB). Integrated mean fluorescence intensity (iMFI) was used to calculate fold changes of phosphorylation. Assay sensitivity was evaluated by comparing phospho flow with Meso Scale Discovery (MSD) and immunohistochemistry (IHC) assays.
Human induced pluripotent cells (iPSCs) were obtained from the HipSci project (http://www.hipsci.org) and differentiated into macrophages using an established protocol (van Wilgenburg, 2013). The genotype_id column of the flow_sample_metadata.txt file contains the canonical HipSci iPSC line name from which the macrophages were differentiated. Data acquisition We used flow cytometry to measure the cell surface expression of three canonical macrophage markers: CD14, CD16 (FCGR3A/FCGR3B) and CD206 (MRC1). Macrophages were cultured in 10 cm tissue-culture treated plates and detached from the plates by incubation in 6 mg/ml lidocaine-PBS solution (Sigma L5647) for 30 minutes followed by gentle scraping. From each cell line we harvested between 300,000-500,000 cells. Detached cells were washed in media, centrifuged at 1200 rpm for 5 minutes and resuspended in flow cytometry buffer (2% BSA, 0.001% EDTA in D-PBS) and split into two wells of a 96-well plate. Nonspecific antibody binding sites were blocked by
Elabscience provides flow cytometry protcols and troubleshootings with elaborate and detailed information. Get the most useful guides for flow cytometry experiment in Elabscience!
Based on long-standing expertise in multicolor flow cytometry, we have compiled a selection of resources that support you in setting up multicolor assays. - Lëtzebuerg
TY - JOUR. T1 - Analysis of nucleocytoplasmic protein shuttling by imaging flow cytometry. AU - Fasler-Kan, Elizaveta. AU - Baiken, Yeldar. AU - Vorobjev, Ivan A.. AU - Barteneva, Natasha S.. PY - 2016. Y1 - 2016. N2 - Many intracellular signal transduction events involve the reversible shuttling of proteins between the cytoplasm and the nucleus. Study of these processes requires imaging information on the protein localization in a given cell and a large number of measurements to obtain sufficient statistics on the protein localization in the whole population. The protocol describes method for quantitative imaging flow cytometry analysis of intracellular distribution of NF-kappaB in ARPE-19 cells stained with specific fluorochrome-conjugated antibodies. The described technique alone or in combination with standard flow cytometry methods can be applied to study any protein undergoing translocation from cytoplasm into the nucleus in a variety of cell lines as well as in heterogeneous primary cell ...
Welcome to the Swanson Biotechnology Center Flow Cytometry Core Facility. The Flow Cytometry Core provides KI and MIT researchers with technical expertise, training and access to sophisticated instrumentation, enabling and supporting the use of a wide range of flow cytometry techniques. This technology allows simultaneous multiparametric analysis of many thousands of cells per second, enabling trained researchers to rapidly analyze complex cell populations using benchtop analysis flow cytometers. High-speed cell sorting services provide researchers with fast, objective and quantitative recording of fluorescent signals from individual cells combined with physical separation of cells of particular interest.. Access is available to all members of the MIT community, to the extent permitted by available capacity. Priority access is given to KI members, NCI-funded research projects and other contributing user groups in recognition of funding support. Depending on available capacity, access may be ...
Our Cellular Senescence Flow Cytometry Assay provides an efficient method to measure Senescence Associated (SA) ß-galactosidase activity. A fluorogenic substrate is added directly to senescent cells in a 35 mm dish. Results can be measured by either flow cytometry or epifluorescence microscope.
The Click-iT Plus EdU Alexa Fluor 350 Flow Cytometry Assay Kit provides a simplified, more robust assay for analyzing DNA replication in proliferating cells compared to traditional BrdU methods. Newly synthesized DNA is analyzed using the UV laser of the flow cytometer. The Click-iT Plus formulation
Browse Flow cytometry products on Labviva. Find relevant scientific protocols, papers and to help find the right product for your application.
Flowcytometry provides an unequalled technique for the analysis and sorting of specific cell types in the context of larger populations of cells. Combined use of defined flow cytometric reagents not only allows to identify minute numbers of specific cells in a total population but also enables to elucidate key aspects of their biological function. Moreover, flow cytometric cell sorting techniques, up to the level of single cell sorting, permit further investigation and modulation of specific cell populations for scientific or clinical purposes under experimentally defined conditions. Since flowcytometry is a technique employed by many researchers, the UMCG has stationed the available flow cytometry equipment in a core facility: the Flow Cytometry Unit (FCU). For research purposes, the FCU accommodates 5 analytical flow cytometers (two FACSCaliburs, one FACSVerse and one LSR-II from BD Biosciences and a MACSQuant from Miltenyi Biotec). For sorting the facility has two high speed multicolor cell ...
Keywords: Flow cytometer, avalanche photodiode, APD near infrared, NIR, sign to noise Intro Flow cytometry is becoming among the fundamental equipment for research of natural systems, as well as the applications and uses of the tools is constantly on the increase in such areas as molecular biology, pathology and immunology. These tools have discovered such energy because they quickly offer quantitative and correlated information regarding multiple guidelines utilized to characterize cells. Movement cytometry systems and fluorescent labeling strategies have identified a huge selection of specific cell phenotypes in human being blood. The recognition and monitoring of the comprehensive cell types offers added to your understanding of oncology fundamentally, immunology, and pathogenesis. The quantitative info content obtainable from movement cytometry comes from the multi-parameter evaluation of the precise phenotype of specific cells. These guidelines are the physical guidelines of ahead scatter, ...
Beginning Monday, May 11, 2020, the Flow Cytometry Core will be operating in Expanded Research Mode. Please find more info here: Flow Cytometry Core Facility Expanded Research Mode Guidelines. The LJI Flow Cytometry Core Facility is one of the largest flow cytometry cores in the San Diego area. The core facility is a full service flow cytometry laboratory that provides investigators with state-of-the-art equipment along with the necessary expertise and services to support cutting-edge research. The core facility offers a wide range of routine and custom flow cytometry, mass cytometry, cell sorting services, data analysis, strategic planning for novel assays, user training as well as full scale research collaborations. More than fee-for-service providers, flow cytometry core staff members are personally invested in the success of experiments and consider themselves temporary extensions of individual labs. The flow cytometry facility has extensive experience in daily instrument calibration and ...
FMO controls are samples that contain all the antibodies you are testing in your experimental samples, minus one of them. When analyzing the minus, or left out parameter in an FMO control, you give yourself a strong negative control to work with. Its a strong negative control because the left out marker in the FMO control allows you to take into account how the other stains in your panel affect the respective minus parameter. Many flow cytometry gates are difficult to define. This is especially true when youre looking at activation markers within a continuum or accounting for the large data spread that occurs when compensating a 10+ color experiment. The only way to convince reviewers that your gate is in the proper place is by using FMO controls. Heres why you need to use FMO controls for any multicolor flow cytometry experiment and how to prepare these controls properly.. Read More ...
We performed serial quantification of the PNH clone size in 38 samples using multiparameter flow cytometry. Granulocytes, monocytes, and RBCs were gated using forward and side scatter as well as lineage-specific markers. The GPI-linked markers fluorescent aerolysin (FLAER), CD55, and CD59 were comparatively evaluated. We also evaluated CD16 on granulocytes and CD14 on monocytes. The sensitivity of detection by each marker was further defined by serial dilution experiments on a flow-sorted sample. Two patients had quantification of their GPI-negative clones before and after allogeneic BMT. ...
Laboratory directors who routinely utilize flow cytometry for at least part of their diagnostic evaluations in leukemias or lymphomas were surveyed by mail. The survey consisted of 12 questions about the flow cytometry procedures used by the laboratory in evaluating leukemias and lymphomas and on the format and content of their official report. It also requested an example of a typical leukemia/lymphoma report and solicited write-in comments about additional important aspects of using flow cytometry to evaluate leukemias and lymphomas not covered by the questionnaire. The goal of the survey, which was sponsored by the Clinical Cytometry Society (CCS), was to document what directors of flow cytometry laboratories currently consider to be the appropriate contents of a clinical leukemia/lymphoma phenotyping analysis and in what manner and detail they report such flow cytometry results to clinicians. The survey indicated that a large number of markers are routinely evaluated to phenotype leukemias ...
Our Mission: To provide introductory flow cytometry video tutorials; a network to connect with experts and collaborators; access to free software tools for multi-color flow cytometry experiment design, data analysis, fluorescence spectra viewing etc.; free Video conferencing and file sharing; provide a platform for flow cytometry experts to host and promote their content, provide flow based and other assay protocols; Flow Cytometry News, Jobs and Event updates, provide vendors a place to review and improve their products.. ...
Purpose: Chronic lymphocytic leukemia (CLL) is currently incurable with standard chemotherapeutic agents, highlighting the need for novel therapies. Overcoming proliferative and cytoprotective signals generated within the microenvironment of lymphoid organs is essential for limiting CLL progression and ultimately developing a cure.. Experimental Design: We assessed the potency of cyclin-dependent kinase (CDK) inhibitor CR8, a roscovitine analog, to induce apoptosis in primary CLL from distinct prognostic subsets using flow cytometry-based assays. CLL cells were cultured in in vitro prosurvival and proproliferative conditions to mimic microenvironmental signals in the lymphoid organs, to elucidate the mechanism of action of CR8 in quiescent and proliferating CLL cells using flow cytometry, Western blotting, and quantitative real-time PCR.. Results: CR8 was 100-fold more potent at inducing apoptosis in primary CLL cells than roscovitine, both in isolated culture and stromal-coculture conditions. ...
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
If you find yourself starting to plan a clinical flow cytometry assay, here are the top 3 issues to think about as you plan your experiment.
This work reviews common flow cytometric methods and applications for the study of bacterial organisms. Flow cytometry is fluorescent method capable of both quantitative and qualitative analysis at the single cell level. It can offer insights about bacterial population dynamics, phenotypic heterogeneity and more. This work features a basic introduction to flow cytometry and presents some of the commonly measured variables, such as viability or membrane potential with an emphasis on the fluorescent probes used to visualize them. The difficulties of adapting flow cytometry to bacterial physiology are discussed, as well as the advantages and disadvantages of the particular probes and methods. Finally, this work seeks to demonstrate the flexibility as well as the shortcomings of flow cytometry using examples of practical applications in basic research, environmental microbiology, biotechnology, clinical practice. Keywords: flow cytometry, microbial subpopulations, fluorescent labelling, bacterial ...
TY - JOUR. T1 - The prognostic value of multiparametric flow cytometry in AL amyloidosis at diagnosis and at the end of first-line treatment. AU - Muchtar, Eli. AU - Jevremovic, Dragan. AU - Dispenzieri, Angela. AU - Dingli, David M. AU - Buadi, Francis K.. AU - Lacy, Martha. AU - Gonsalves, Wilson. AU - Hayman, Suzanne R.. AU - Kapoor, Prashant. AU - Leung, Nelson. AU - Russell, Stephen J. AU - Lust, John A.. AU - Lin, Yi. AU - Go, Ronald S.. AU - Chakraborty, Rajshekhar. AU - Zeldenrust, Steven. AU - Kumar, Shaji K. AU - Kyle, Robert A.. AU - Rajkumar, S Vincent. AU - Gertz, Morie. PY - 2017/1/5. Y1 - 2017/1/5. N2 - Multiparametric flow cytometry (MFC) in amyloid light-chain (AL) amyloidosis has not been widely adopted and, consequently, there is little information on its clinical relevance. We studied 173 patients with AL amyloidosis who underwent MFC immunophenotyping of bone marrow sample at diagnosis and 82 patients at the end of the first line of treatment (EOT). The number of monotypic ...
Central Department of Biotechnology, Tribhuvan University is organizing International Workshop on Applications of FLOW CYTOMETRY on Biotechnology -2017 on September 14-17, 2017.. For detail information, please click the linked below:. Download Application form and send the filled application form to This email address is being protected from spambots. You need JavaScript enabled to view it.. or submit to Central Department of Biotechnology, Tribhuvan University, Kirtipur. Detail Information. ...
Background: Invariant natural killer cells (iNKT) are an important immunoregulatory T cell subset. Currently several flow cytometry-based approaches exist for the identifi-cation of iNKT cells, which rely on using the 6B11 monoclonal antibody or a combina-tion of anti-Vα24 and anti-Vβ11 antibodies. Objective: The aim of this study was to compare the ability of two flow cytometry-based methods for detecting the frequency of circulating iNKT cells. Methods: The frequency of iNKT cells was detected in the pe-ripheral blood of 37 healthy adult donors by flow cytometry using the 6B11 antibody or a combination of anti-Vα24 and anti-Vβ11 antibodies. Results: The frequency of iNKT cells detected by 6B11 antibody or by combination of anti-Vα24 and anti-Vβ11 anti-bodies was significantly different (0.54% vs. 0.31%, respectively, p|0.001) but the val-ues were highly correlated (Spearman r = 0.742, p|0.0001). Conclusion: The results of this study indicate that different combinations of mAbs detect different
Plasmodium vivax preferentially infects Duffy-positive reticulocytes and infections typically have few parasite-infected cells in the peripheral circulation. These features complicate detection and quantification by flow cytometry (FC) using standard nucleic acid-based staining methods. A simple antibody-based FC method was developed for rapid parasite detection along with simultaneous detection of other parasite and erythrocyte markers. Clinical samples were collected from patients diagnosed with P. vivax at a district Malaria Clinic in Kanchanaburi, Thailand. One μL of infected blood was washed, fixed, stained with a Plasmodium pan-specific anti-PfBiP antibody conjugated with Alexa Fluor 660, and analysed by FC. Additional primary conjugated antibodies for stage-specific markers of P. vivax for late trophozoite-early schizonts (MSP1-Alexa Fluor 660), late-stage schizonts (DBP-Alexa Fluor 555), and sexual stages (Pvs16) were used to differentiate intra-erythrocytic developmental stages. The
Multiple myeloma (MM) is a hematological plasma cell malignancy in the bone marrow. Lately, increased knowledge of MM pathogenesis and advances in therapy have improved the survival of MM patients. However, due to the unique and complex genome of each patient, some patients are resistant to standard therapies while others achieve durable response but eventually experience relapse. Therefore, new strategies especially for relapsed and refractory and high-risk multiple myeloma (RRMM, HRMM) patients, who have poor response to current therapies, are required. Melflufen, a novel prodrug of the alkylating agent melphalan, has shown significantly decreased resistance effects and more selective cytotoxicity compared to melphalan in vitro and in vivo, but the molecular markers identifying the sensitive subgroups of MM patients have not yet been discovered. The aim of this study was to identify a melflufen-sensitive subgroup of MM patients by utilizing a high throughput flow cytometry-based drug ...
Flow cytometry immunophenotyping is invaluable for the diagnostic and prognostic work-up of haematological malignancies. Over the last decade, multi-colour flow cytometry analysis has been used increasingly to investigate leptomeningeal disease; several studies show the utility and sensitivity of this technique for both B cell Non-Hodgkins Lymphoma and Acute Leukaemia diagnostics. Cells within CSF samples are typically low in number and degrade quickly. Therefore, the recommendation is to use cell stabilisation reagents for CSF samples that require flow cytometry analysis. Previous studies have shown that TransFix stabilised CSF samples have a cell yield equal to or better than that of fresh CSF samples and so may be used for immunophenotypic analysis after 18 hours of storage. Some cell characteristics may change after TransFix treatment: for example, light scatter properties of peripheral blood and CSF leukocytes may be altered and the fluorochrome signal intensity of certain bound antisera ...
p style=margin: 0cm 0cm 0pt; text-align: justify; line-height: normal;,Introduction: Medicinal plants are considered to be safer, non-toxic and less harmful as compared to synthetic based drugs that are available. In this study, we focused on aqueous leaves extract of Calotropis gigantea for determining its antimicrobial activity in infected (dengue) human whole blood samples using flow cytometry.Methods: Infected dengue human blood samples (n = 5; confirmed on the basis of NS1 antigen to dengue virus;) were collected from pathology lab and evaluated its blood counts (lymphocytes, monocytes and granulocytes count); forward scatter (FSC) and side scatter (SSC) including CD14 monocyte surface marker following the use of variable doses of aqueous leaves extract of C. gigantea.Results: In this study, the results showed that aqueous leaves extract of C. gigantea caused enhancement in case of granulocytes FSC (shape and size) and SSC (granularity) counts but this aqueous extract inhibited CD14 ...
TY - JOUR. T1 - Flow cytometric methods for the analysis of human basophil surface antigens and viability. AU - Bochner, Bruce S.. AU - McKelvey, Alicia A.. AU - Schleimer, Robert P.. AU - Hildreth, James. AU - MacGlashan, Donald W.. PY - 1989/12/20. Y1 - 1989/12/20. N2 - Fluorescence and flow microfluorometric methods have been established for the detection and evaluation of IgE-bearing human leukocytes in various cell preparations including those where basophils are present at low percentages. Quantitative techniques for the determination of basophil purity, viability, and cell surface antigens including IgE are described. Use of these methods will facilitate the identification and phenotypic analysis of human IgE-bearing cells in a wide variety of biological fluids.. AB - Fluorescence and flow microfluorometric methods have been established for the detection and evaluation of IgE-bearing human leukocytes in various cell preparations including those where basophils are present at low ...
TY - JOUR. T1 - Multicolor flow-cytometric analysis of milk allergen-specific T-helper type 2 cells revealed coexpression of interleukin-4 with Foxp3. AU - Yamawaki, Kazuo. AU - Inuo, Chisato. AU - Nomura, Takayasu. AU - Tanaka, Kenichi. AU - Nakajima, Yoichi. AU - Kondo, Yasuto. AU - Yoshikawa, Tetsushi. AU - Urisu, Atsuo. AU - Tsuge, Ikuya. N1 - Funding Information: Funding Sources: This study was supported in part by the Health and Labor Sciences Research Grants of the Research on Allergic Disease and Immunology from the Ministry of Health, Labour and Welfare of Japan (to U.A. and I.T.). Publisher Copyright: © 2015 American College of Allergy, Asthma & Immunology.. PY - 2015/12/1. Y1 - 2015/12/1. N2 - Background Allergen-specific T-helper type 2 (TH2) cells play an important role in the development of allergic inflammation; however, investigations of the properties of allergen-specific T cells have been challenging in humans. Despite clear evidence that forkhead box p3 (Foxp3) is expressed ...
The ring-stage survival assay (RSA) is a powerful tool for phenotyping artemisinin-resistant Plasmodium falciparum but requires experienced microscopists to count viable parasites among 10,000 erythrocytes in Giemsa-stained thin blood smears. Here we describe a rapid flow cytometric assay that accurately counts viable parasites among 250,000 erythrocytes in suspension. This method performs as well as light microscopy and can be used to standardize the collection of RSA data between research groups in laboratory and field settings.
Methods have been developed for isolating human tissue macrophages from first trimester or term pregnancy decidua. After a two stage enzymic digestion, viable cells were separated from cellular debris by velocity sedimentation at unit gravity or by Percoll centrifugation. Cell populations were analysed by flow cytometry after labelling with monoclonal antibodies. In term decidua, 47% of the cells were of bone marrow origin, comprising 18% macrophages, 3% large granular lymphocytes and 8% T cells. The remaining cells, the proportion of which varied between individuals, were CD16-positive granulocytes. Macrophages were isolated flow cytometrically from both first trimester and term decidual cell dispersions after labelling with an antibody to MHC class II. Yields of up to 4 X 10(6) macrophages, greater than 95% pure, were routinely obtained.
This is a page with useful consensus documents and standards for people interested in clinical flow cytometry. The standards are listed in alphabetical order . If we are missing something, please feel free to contribute by adding it here. 2006 Bethesda International Consensus Recommendations on Immunophenotypic Analysis of Hematolymphoid Neoplasia by Flow Cytometry. Cytometry Part B, 2007. 2006 Bethesda International Consensus Recommendations on the Immunophenotypic Analysis of Hematolymphoid Neoplasia by Flow Cytometry: Recommendations for Training and Education to Perform Clinical Flow Cytometry, Cytometry Part B 2007 Clinical Flow Cytometric Analysis of Hematolymphoid Cells; Approved Guideline - Second Edition H43-A2 Clinical and Laboratory Standards Institute (CLSI) 2007 Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline-Second Edition H42-A2 Clinical and Laboratory Standards Institute (CLSI) 2007. MMWR 2003: Guidelines for Performing ...
This vignette will guide you through analysis of an example flow cytometry data set from an experiment examining time-lapse florescence reporter levels from a synthetic biological circuit in liquid cultures of budding yeast. In this example circuit, fluorescent reporter expression is mediated by a transcription factor/transcriptional repressor complex. The transcriptional repressor is degraded via the ubiquitin proteasome system, in response to a small molecule. Fluorescence levels are measured approximately every 10 minutes by flow cytometry. Here we demonstrate how to import the resulting .fcs files into R, gate and annotate this data with experimental metadata (e.g. the ...
Phenotype and functional heterogeneity of airway smooth muscle (ASM) cells in vitro is well known, but there is limited understanding of these features in vivo. We tested whether ASM is composed of myocyte subsets differing in contractile phenotype marker expression. We used flow cytometry to compare smooth muscle myosin heavy chain (smMHC) and smooth muscle-alpha-actin (sm-alpha-actin) abundance in myocytes dispersed from canine trachealis. Based on immunofluorescent intensity and light scatter characteristics (forward and 90 degrees side scatter), 2 subgroups were identified and isolated. Immunoblotting confirmed smMHC and sm-alpha-actin were 10- and 5-fold greater, respectively, in large, elongate myocytes that comprised -60% of total cells. Immunohistochemistry revealed similar phenotype heterogeneity in human bronchial smooth muscle. Canine tracheal myocyte subpopulations isolated by flow cytometry were used to seed primary subcultures. Proliferation of subcultures established with myocytes ...
The XN-20, is a full blood count (FBC) analyser with an extended differential counting and flagging System. The XN-Series individual channels allow real-time reflex analysis, and uses a two stage process to classify the white blood count (WBC) sub-populations and detect the presence of abnormal reactive and malignant cells. In regards to lymphocytes in the peripheral blood, the machine has the capacity to distinguish activated from non-activated T-cell subsets using a very small volume of EDTA sample (88uL) (including remnant sample from a standard full blood count) with results available in 1.5 minutes. It is a fully automated process and can be considered as an alternative rapid flow cytometry method.. Objective of the SASA study: to investigate the signal pattern of white blood cells assessed using the XN-20 full blood count platform in patients with untreated viral infections i.e. HIV, HCV and HBV. The data from the analysis will be reviewed in conjunction with patients demographic and ...
Transplantation improves the health and quality of life for patients suffering from renal failure, but for many, antibodies specific for HLA antigens create a substantial barrier. Our center has developed a desensitization protocol to remove HLA-specific antibodies and successfully transplant these sensitized patients. This protocol involves alternate day plasmapheresis, intravenous immunoglobulin (IvIg), and immunosuppressants. We use a Luminex assay to detect HLA-specific antibody and developed a flow cytometric technique using HLA tetramer molecules to quantitate HLA-specific B cells. Following desensitization and transplantation, we observe a sustained loss of donor-specific HLA antibodies, while antibodies specific for 3rd party HLA return to pre-treatment levels. Interestingly, B cells specific for donor-specific HLA persist. We hypothesize that transplantation in the absence of inflammation, due to continued desensitization treatment, allows for the induction of B cell anergy toward ...
Find an overview of recombinantly generated REAfinity antibodies and flow cytometry data for improved immune checkpoint detection. - Liechtenstein
Background The frequency of intraperitoneal free tumor cells (IPTC) is considered to reflect the severity of peritoneal metastasis (PM). We quantified the relative number of IPTC against leukocytes in...
Global Flow Cytometry Market is expected to reach USD 8.0 billion by 2025, according to a new study by Grand View Research, Inc. The increasing incidence of infectious diseases and cancer is expected to upsurge the demand for flow cytometers for use in disease diagnosis over the coming years. In addition, higher number of physicians is inclined toward the usage of autologous and allogenic stem cell therapy, due to adverse effects caused by chemotherapy & radiation therapy in the treatment of cancer, thereby affecting the growth of this market.. Moreover, rising demand for point-of-care testing in chronic diseases management is expected to fuel the demand for cytometry techniques. Rising implementation of microfluidic miniature flow cytometry in point-of-care diagnostics is the factor augmenting the future growth. Increasing R&D initiatives by various key players for the development of multicolor assays and advanced reagents for analysis are anticipated to boost the usage rate. Advancement in ...
Our understanding of heme sensing and the regulation of heme-iron acquisition by fungal pathogens is incomplete. Heme contains ∼80% of the iron in vertebrate hosts, and we previously demonstrated that heme is an important iron source for C. neoformans (35, 74). In the present study, we constructed a strain encoding a cytosolic heme sensor to address our goals of (i) understanding heme-iron acquisition during fungal proliferation and pathogenesis and (ii) identifying potential heme/iron-related targets for antifungal therapy. Initially, we validated the behavior of the heme sensor by demonstrating responsiveness to exogenous hemin by established microscopic, fluorimetric, and flow cytometry methods (41). Importantly, we demonstrated that the sensor detected the reduction in cytosolic heme levels expected to result from impaired endocytosis. Specifically, reduced cytosolic heme levels were found in cells treated with CPZ and in mutants such as the cig1 and las17 mutants that have lower heme ...
Defining Human Dendritic Cell Progenitors by Multiparametric Flow Cytometry. G. Breton, J. Lee, K. Liu, M.C. Nussenzweig: Defining human dendritic cell progenitors by multiparametric flow cytometry. Nat Protoc. 2015 Sep;10(9):1407-22. PMC4607256.. Dendritic cells (DCs) play an important role in the maintenance of mucosal homeostasis and the induction of antigen-specific mucosal immune responses. DCs develop from bone marrow from increasingly restricted but phenotypically well-defined progenitor […]. ...
There are times a flow cytometry study is performed and is inconclusive, so immunohistochemical (IHC) stains are done. Can they be billed together? Yes, you can bill them in some circumstances with proper documentation but there are specific guidelines on the billing of these services.
The Flow Cytometry Core Facility provides a broad range of equipment and services for the analysis and isolation of cells and other similar-sized particles based on fluorescent labeling. Four high-speed cell sorters are available to enable optimized sorting of highly purified cells, subcellular organelles or bacteria. The facility also supports a variety of analytical flow cytometers capable of performing basic to advanced multiparameter fluorescence analysis of many types of cell suspensions as well as analysis of intracellular signaling. An experienced staff of technical experts provides instruction in the design, execution and analysis of flow cytometry-based studies. ...
LifeLabs is excited to introduce an expansion of our flow cytometry testing. Starting on April 10th, 2017 we will offer flow cytometric immunophenotyping for hematopoietic and lymphoid malignancies in addition to our existing flow cytometry testing for T-cell subset analysis.. Flow cytometry is widely used for analyzing the expression of surface and intracellular molecules in order to differentiate and characterize different cell populations. It continues to be a necessary diagnostic tool for the classification, staging, and monitoring of hematolymphoid neoplasms.. LifeLabs is pleased to offer a variety of panels to facilitate the diagnosis and/or prognosis of the following:. ...
Tercen is technology start-up with a distributed working model that has people working full-time and part-time. We have developed a software platform aimed at supporting bioinformaticians with automation tools, and helping non-coding scientists get up the data-analysis learning curve.. www.tercen.com. We are looking for a Data Scientist, Software Developer, or Bioinformatician who has experience with computational biology for Flow Cytometry. The purpose is to have the candidate code plug-ins for our software that focus specifically on analysis of Flow Cytometry data.. The job will include R implementation of bioinformatics algorithms R-Shiny applications Documentation of the software life-cycle. Scientific domain knowledge in Flow Cytometry is a requirement. The successful candidate ideally will have two years bioinformatics experience in this field. Alternately, it is acceptable for a candidate (with strong coding skills) to have experience working with Flow Cytometry laboratory equipment in a ...
Flow cytometry (FCM) bioinformatics is a sub-field of bioinformatics, aimed at developing effective and efficient computational tools to store, organize, and analyze high-throughput/dimensional FCM data. Flow cytometers are capable of analyzing thousands of cells per second for up to 40 features. These features primarily signal the presence of different proteins on cells in the bloodstream. Hence contributing large amounts of data towards the big biological data paradigm. The data that a flow cytometer outputs from a biological sample, is called a FCS file.The International Mouse Phenotyping Consortium (IMPC) is a collaboration between 23 international institutions and funding organizations. Its aim is to decipher the function of 20,000 mouse genes. IMPC is doing so by breeding mice with a certain gene knocked out (KO), cancelling the function of that gene. In turn, FCM is used to measure the immunological changes correlated to this knockout. Many tools exist to classify FCS files. However, ...
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Chinese hamster ovary cells were seeded in the absence or presence of the polyamine synthesis inhibitor 2-difluoromethylornithine (DFMO). At 1-4 days after seeding, the cells were labelled for 15-120 min with the thymidine analogue bromodeoxyuridine (BrdUrd) and they were then fixed directly after the labelling period. In addition, cells were labelled for 30 min and they were then allowed to progress in BrdUrd-free medium during a defined post-labelling time before fixation. An indirect immunofluorescence technique, using the monoclonal BrdUrd antibody and the intercalating stochiometric DNA stain, propidium iodide, was applied to enable quantification of cellular BrdUrd and DNA contents, respectively, by flow cytometry (FCM). By comparing the mean DNA content of BrdUrd-labelled cells to the mean DNA contents of G(1) and G(2) cells, a relative measure of the position of the BrdUrd-labelled cells was obtained (relative movement). Relative movement data, obtained from control and DFMO-treated ...
Scientists from the Department of Horticulture are using modern laser-based flow cytometry techniques for isolating plant protoplasts. Such applications are at the leading edge in this field. The approach allows identification and physical isolation of single plant chromosomes. ...
Introduction: We have reported previously that α-myosin heavy chain (α-MyHC) expressing myocytes (MCs), the predominant MC in adult mouse hearts, hypertrophy under pressure-overload without a proportionate increase in total MyHC protein (T-MyHC) content (Lopez et al, Circ. Res., 2011). It is not yet known if during normal physiological growth, these MCs increase their T-MyHC content in proportion to cell size.. Hypothesis: During normal post-natal growth, an increase in T-MyHC content is proportionate to changes in cardiac mass and MC size.. Methods: Individual cardiac cells were isolated by enzymatic digestion from male C57Bl/6 mice age in days 22+/-1 (young, n=4) and 88+/-6 (adult, n=4). Body and heart weights (wt), mean MC volumes by Coulter Multisizer (vol), and cell protein content-per-MC by BCA assay (TotProtMC) were used to measure cardiac growth. A new approach using large-particle fluorescent activated cell sorting (FACS, see Figure) was validated to isolate 15K Troponin-T expressing ...
The effect of single-drug cis-diamminedichloroplatinum (CDDP) treatment on the distribution of T-lymphocyte subsets and monocytes was determined using monoclonal antibodies and a flow cytometry technique. Thirty-nine patients with radically operated ovarian carcinoma received postoperative treatment with six cycles of CDDP 50 mg/m2, and were examined either before, during, or after chemotherapy. During treatment, the number of OKT4+ (mainly T-helper) cells was reduced, whereas the number of OKT8+ (mainly T-suppressor/cytotoxic) cells was increased. Four to six months after the CDDP treatment was ended, these aberrations were no longer seen. The number of 1D5+ cells (monocytes) was not influenced by the treatment. It is concluded that no long-lasting change in the distribution of immunocompetent cells could be detected after this regimen of CDDP treatment.
The Aurora family of serine-threonine kinases are essential regulators of cell division in mammalian cells. Aurora-A and -B expression and kinase activity is elevated in a variety of human cancers and is associated with high proliferation rates and poor prognosis. AMG 900 is a highly potent and selective pan-aurora kinase inhibitor that has entered clinical evaluation in adult patients with advanced cancers. In mice, oral administration of AMG 900 blocks the phosphorylation of histone H3 on serine-10 (p-Histone H3), a proximal substrate of aurora-B and inhibits the growth of multiple human tumor xenografts, including multidrug-resistant models. In order to establish a preclinical pharmacokinetic-pharmacodynamic (PK-PD) relationship for AMG 900 that could be translated to the clinic, we used flow cytometry and laser scanning cytometry detection platforms to assess the effects on p-Histone H3 inhibition in terms of sensitivity, precision, and specificity, in human tumor xenografts in conjunction with
Written by Tim Bushnell, Ph.D. Flow cytometry is a powerful technique impacting both clinical and research.. If youre looking for a career, flow cytometry technology can take you many places. An experienced flow cytometrist can find a job in a biotechnology company, in academia, or in a clinical setting.. Rather than focusing on specific career information, I would like to highlight one career option that has been very good for me since my transition from research scientist to core manager over a decade ago.. Core facilities, or Shared Resource Laboratories (SRL) as this Cytometry A paper made popular, represent an investment by institutions in resources and personnel.. Staff and directors working in SRLs have the advanced training and experience to support the researchers and research mission of the institution.. Working in an SRL is a different and very exciting career option for researchers who enjoy flow cytometry, enjoy working on many different projects, and enjoy working in scientific ...
Summer Research Description: Dynein is an essential cytoskeletal motor protein, facilitating the transport of various materials within the cell by binding to and trafficking along microtubules. In non-dividing cells, dynein transports cargo from the cell periphery towards the nucleus and microtubule-organizing center, in a microtubule minus-end directed manner. Previous data indicates that dynein plays a role in HIV intracytoplasmic transport. In these studies, I used flow cytometry and X-gal analysis to assess the effects of Ciliobrevin, an inhibitor of dynein ATPase activity, on HIV infectivity. Using flow cytometry, I found that increasing the concentration of Ciliobrevin, gives an overall decrease in infectivity, however not to the extent shown by previous studies, likely due to differences in the approaches used to quantify the extent of infection. This increased inhibition of dynein leads to lowered infectivity because fewer HIV particles are being trafficked to the nucleus. I am also ...
Male mice model of degenerative liver was obtained through food fasting but still have drinking water for 5 days. It caused energy protein malnutrition and damage of liver tissue. The administration of 50% (v/v) honey was performed for 10 consecutive days, while the positive control group was fasted and not given honey and the negative control not fasted and without honey. Observations of regeneration the liver tissue based on histologically examination, observation of Hsp70 expression, and homing signal based on vascular endothelial growth factor-1 (VEGF-1) expression using immunohistochemistry technique. Observation on expression of CD34 and CD45 as the marker of auto mobilization of hematopoietic stem cells using flow cytometry technique ...
Malik SA, Orhon We, Morselli E, Criollo A, Shen S, Marino G, BenYounes A, Benit P, Rustin P, Maiuri MC, Kroemer G. of level of resistance. Outcomes Induction of apoptosis in principal FL cells after venetoclax treatment Venetoclax treatment induced a focus C dependent reduction in cell viability in six FL principal samples (Amount ?(Figure1A).1A). The GANT 58 LY78 test was the most delicate (IC50 = 11 nM) as well as the LY97 test one of the most resistant (IC50 > 200 nM) to venetoclax treatment. To see upon the number of venetoclax replies noticed, we motivated the appearance of BCL-2 and BIM in major FL examples by movement cytometry [10] (Body ?(Figure1B).1B). Following flow cytometric evaluation of BCL-2 and BIM amounts revealed a substantial (positive cells(A) Apoptosis induction in major FL examples after venetoclax treatment. Major cells had been treated with venetoclax for 4 H and Annexin-V/7-AAD structured movement cytometry assay was performed to look for the percentage of ...
Flow cytometry; Cytofluorometry, Flow; Cytometry, Flow; Microfluorometry, Flow; Flow Microfluorimetry; Fluorescence-Activated Cell Sorting. On-line free medical diagnosis assistant. Ranked list of possible diseases from either several symptoms or a full patient history. A similarity measure between symptoms and diseases is provided.
I am a marine biologist and have worked at the International Research Institute of Stavanger (IRIS) for thirteen years. My main research interest is investigating effects of environmental changes on the physiology and behaviour of marine invertebrates. I have worked primarily with decapod crustaceans and bivalve molluscs, examining, for example, cardiac performance, immune function, valve gaping behaviour and expression of endogenous rhythms. A key component of this work is the development of methods that produce results readily interpreted within an ecological context. I am also involved in the development of flow cytometry techniques to measure blood cell parameters in samples taken from blue mussels. I have recently led projects tasked with developing monitoring techniques to detect oil discharges at sea, together with an investigation into potential environmental impacts arising from leakage of carbon dioxide from sub-sea storage. I am currently working on a project that aims to use the ...
The hallmark of an immune response to Mycobacterium tuberculosis is granuloma formation. CD4+Th1 cells are of central importance in anti-mycobacterial immunity by way of producing effector IFN-γ and TNF-α cytokines that help to contain infection in the granulomas by activating macrophages. The environment of the granuloma is hypoxic and we have little knowledge on CD4+ T cell processes and function in this environment. Autophagy is a de novo intracellular protein and organelle degradation pathway induced in eukaryotic cells by stress-factors such as starvation or hypoxia. It has been described that autophagy is induced in T cells upon activation and autophagic processes might contribute to nutrient supply and the regulation of cytokine production in T cells. The aim of this study was to investigate autophagic processes in CD4+ T cells after activation and the regulation of IFN-γ production under normoxia, and hypoxia as found in the granuloma of TB patients. The main assays, immunoblotting ...
Purpose:. Circulating tumor cell (CTC) enumeration or Androgen Receptor (AR) splice variant 7 expression from Epithelial Cell Adhesion Molecule (EpCAM) positive CTCs are predictive biomarkers for patients with prostate cancer. Recent reports suggest subpopulations of CTCs with decreased EpCAM expression may have greater invasive capacity or drug-resistant potential. These include cells with mesenchymal phenotypes that express N-cadherin, or MUC1, or stem cell populations that express CD133. Integrated molecular analysis across different CTC subpopulations has not been performed. We sought to compare the molecular profile of different populations of CTCs from the blood of patients with prostate cancer.. Methods:. A multiparametric flow cytometry assay was used to identify different populations of CTCs from patients with prostate cancer. We then employed an integrated CTC capture and analysis technology known as the VERSA (Versatile Exclusion-based Rare Sample Analysis) platform to in parallel or ...
Dec 09, 2020 (Heraldkeepers) -- The Global Flow Cytometry Market is segmented on the lines of its technology, component, application, end user and regional. Based on technology segmentation it covers bead based flow cytometry and cell based flow cytometry. Under component segmentation it covers instruments which further segmented into cytometry platforms, replaceable components, accessories, reagents and consumables, software and services. Based on application segmentation it covers academic and clinical research applications and diagnosis applications. Under end user it consists of commercial organizations, medical schools and clinical labs, hospitals, academics and others. The Global Flow Cytometry Market on geographic segmentation covers various regions such as North America, Europe, Asia Pacific, Latin America, Middle East and Africa. Each geography market is further segmented to provide market revenue for select countries such as the U.S., Canada, U.K. Germany, China, Japan, India, Brazil, ...
Product list of China Fcm Flow Cytometry System, show the variety of China products related to Fcm Flow Cytometry System; You can choose the right product of China Fcm Flow Cytometry System on this list.
This video demonstrates how to harvest cells from a spleen sample, and prepare a single cell suspension prior to performing cell isolation. Preparing a true single cell suspension of the primary tissue sample will optimize cell separation by avoiding addi
Background: CD34 a 110- to 115-Kda transmembrane sialoglyco pro- V twin, is expressed by early hematopoietic (myeloid and lymphoid) progenitor cells, endothelial cells, murine embryonic fibroblasts, and bone marrow (BM) stromal cells and their precursors. CD34 expression has great importance in diagnosis, classification and management of acute leukemia. Aim of the study: To evaluate the expression of CD34 (percentage and meanfluorescence intensity) in Sudanese patient with acuteleukemia. Materials and Methods: This is descriptive cross-sectional study, conducted in Flowcytometry centre in Khartoum state, 50 new cases of acute leukemia analyzed by flowcytometry for CD34expression. The flowcytometery lysing procedure for bone marrow aspiration and peripheral blood monoclonal antibody combination was done as the follows: All tubes were labeled then pipette into each tube 20 µL of CD34 monoclonal antibody and added 100 µL of sample containing no more than 1 x 104 leukocytes / ml. The tubes were ...
A procedure for enrichment in recombinant interleukin-2 (rIL2)-activated natural killer (NK) cells was developed and used for in vitro generation of antitumor effector cells from the peripheral blood of 20 patients with central nervous system (CNS) tumors. In comparison to the patients unseparated mononuclear cells and nonadherent lymphocytes cultured in the presence of 1000 U/ml of rIL2 for up to 3 weeks, interleukin-2-stimulated lymphoid cells, when purified by adherence to plastic, proliferated better (up to 6,720-fold expansion) and achieved up to five times higher levels of antitumor activity against K562 cell targets and NK-resistant glioblastoma cell targets. Two-color flow cytometry analysis showed that cultures of cells purified by adherence to plastic which had the best proliferation contained 10% or less of CD3+Leu19-T-lymphocytes, while the unseparated lymphokine-activated killer cell cultures which proliferated poorly contained up to 85% of CD3+Leu19-T-cells. Cultures of adherent ...