A5041629116- Single-cell and spatial transcriptomics
- Extracellular vesicles in disease
- Immune Cell Function and Interaction
- T-cell and B-cell Immunology
- Cell Image Analysis Techniques
- MicroRNA in disease regulation
- Dermatology and Skin Diseases
- Asthma and respiratory diseases
- Allergic Rhinitis and Sensitization
- Gene expression and cancer classification
- Immune cells in cancer
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Bacterial Infections and Vaccines
- Microfluidic and Bio-sensing Technologies
- Advanced Biosensing Techniques and Applications
- Lattice Boltzmann Simulation Studies
- Gene Regulatory Network Analysis
- Microscopic Colitis
- Metabolomics and Mass Spectrometry Studies
- Advanced Fluorescence Microscopy Techniques
- Food Allergy and Anaphylaxis Research
- Scientific Computing and Data Management
- Helicobacter pylori-related gastroenterology studies
- Mitochondrial Function and Pathology
- Inflammatory Bowel Disease
National Institutes of Health
2023
University of Ottawa
2023
National Cancer Centre Japan
2023
National Cancer Research Institute
2023
Center for Cancer Research
2023
University of Virginia
2010-2021
Virginia Commonwealth University Medical Center
2016
University of Virginia Health System
2008-2015
City Of Hope National Medical Center
2004
ABSTRACT Extracellular vesicles (EVs) are small, heterogeneous and difficult to measure. Flow cytometry (FC) is a key technology for the measurement of individual particles, but its application analysis EVs other submicron particles has presented many challenges produced number controversial results, in part due limitations instrument detection, lack robust methods ambiguities how data should be interpreted. These complications exacerbated by field's reporting framework, EV‐FC manuscripts...
Extracellular vesicles (EVs) mediate targeted cellular interactions in normal and pathophysiological conditions are increasingly recognised as potential biomarkers, therapeutic agents drug delivery vehicles. Based on their size biogenesis, EVs classified exosomes, microvesicles apoptotic bodies. Due to overlapping ranges the lack of specific markers, these classes cannot yet be distinguished experimentally. Currently, it is a major challenge field define robust sensitive technological...
This 40-color flow cytometry-based panel was developed for in-depth immunophenotyping of the major cell subsets present in human peripheral blood. Sample availability can often be limited, especially cases clinical trial material, when multiple types testing are required from a single sample or timepoint. Maximizing amount information that obtained not only provides more characterization immune system but also serves to address issue limited availability. The presented here identifies CD4 T...
Microparticles (MPs) are submicron vesicles released from cell membranes in response to activation, injury, or apoptosis. The clinical importance of MPs has become increasingly recognized, although no standardized method exists for their measurement. Flow cytometry (FCM) is the most commonly used technique, however, because small size MPs, and limitations current FCM instrumentation, accurate identification compromised by this methodology. We decided investigate whether use combined with...
We have shown that the 12/15-lipoxygenase (12/15-LO) product 12<i>S</i>-hydroxyeicosatetraenoic acid increases monocyte adhesion to human endothelial cells (EC) <i>in vitro</i>. Recent studies implicated 12/15-LO in mediating atherosclerosis mice. generated transgenic mice on a C57BL/6J (B6) background modestly overexpressed murine gene (designated LOTG). LOTG had 2.5-fold elevations levels of and 2-fold increase expression protein vivo</i>. These developed spontaneous aortic fatty streak...
Fluorescence activated cell sorting is the technique most commonly used to separate primary mammary epithelial sub-populations. Many studies incorporate this before analyzing gene expression within specific cellular lineages. However, our knowledge, no one has examined effects of fluorescence (FACS) separation on short-term transcriptional profiles. In study, we isolated a heterogeneous mixture cells from mouse gland. To determine isolation and process expression, harvested RNA enzymatic...
Abstract Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and urogenital diseases. Centrifugation is the most common method used to enrich uEVs. However, a majority of studies date have focused on ultracentrifugation pellet, potentially losing novel source important biomarkers that could be obtained at lower centrifugation. Thus, aim this study rigorously characterize first time uEVs in low speed pellet determine minimal volume urine required proteomic analysis (≥9.0 mL...
Abstract Technological advancements in fluorescence flow cytometry and an ever‐expanding understanding of the complexity immune system have led to development large panels reaching up 43 colors at single‐cell level. However, as panel size increase, so too does detail involved designing optimizing successful high‐quality fit for downstream high‐dimensional data analysis. In contrast conventional cytometers, full‐spectrum cytometers measure entire emission spectrum each fluorophore across all...
High-dimensional single-cell data has become an important tool in unraveling the complexity of immune system and its involvement homeostasis a large array pathologies. As technological tools are developed, researchers adopting them to answer increasingly complex biological questions. Up until recently, mass cytometry (MC) been main technology employed cytometric assays requiring more than 29 markers. Recently, however, with introduction full spectrum flow (FSFC), it possible break...
Abstract Budding yeast Saccharoymyces cerevisiae is a powerful model system for analyzing eukaryotic cell cycle regulation. Yeast analysis typically performed by visual or flow cytometry, and both have limitations in the scope accuracy of data obtained. This study demonstrates how multispectral imaging cytometry (MIFC) provides precise quantitation distribution morphological phenotypes cells flow. Cell wild‐type yeast, nap1Δ , overexpressing NAP1 was visually, MIFC. Quantitative employed...
For an emerging disease like COVID-19, systems immunology tools may quickly identify and quantitatively characterize cells associated with progression or clinical response. With repeated sampling, immune monitoring creates a real-time portrait of the reacting to novel virus before disease-specific knowledge are established. However, single cell analysis can struggle reveal rare that under 0.1% population. Here, machine learning workflow Tracking Responders EXpanding (T-REX) was created...
Flow cytometry (FCM) is a common method for characterizing extracellular particles (EPs), including viruses and vesicles (EVs). Frameworks such as MIFlowCyt-EV exist to provide reporting guidelines metadata, controls, data reporting. However, tools optimize FCM EP analysis in systematic quantitative way are lacking. Here, we demonstrate cohesive set of methods software that settings facilitate cross-platform comparisons studies. We introduce an automated small-particle optimization (SPOT)...
Abstract The need for more in‐depth exploration of the human immune system has moved flow cytometry field forward with advances in instrumentation, reagent development and availability, user‐friendly implementation data analysis methods. We developed a high‐quality 45‐color panel, comprehensive characterization major cell lineages present circulation including T cells, γδ NKT‐like B NK monocytes, basophils, dendritic ILCs. Assay optimization steps are described detail to ensure that each...
The purpose of this document is to define minimal standards for a flow cytometry shared resource laboratory (SRL) and provide guidance best practices in several important areas. This effort driven by the desire International Society Advancement Cytometry (ISAC) members SRLs maintain excellence cytometry, act as repository key elements information (e.g. example SOPs/training material, etc.). These are not intended specifically how implement these recommendations, but rather establish goals an...
Abstract Modern flow cytometric cell sorters are all capable of so‐called “high‐speed sorting.” However, there is confusion about exactly how fast a “high‐speed” sorter can sort cells. There many considerations in achieving the fastest sorting speed, as well highest quality results—cell recovery, purity, and functionality. This requires same required for “slow‐speed sorting”; however, more precise implementation high‐speed sorting. The modern enable because advances electronics data...
It is not understood why only some infections with Entamoeba histolytica result in disease. The calcium-regulated transcription factor upstream regulatory element 3-binding protein (URE3-BP) was initially identified by virtue of its role regulating the expression two amebic virulence genes, Gal/GalNac lectin and ferredoxin. Here we tested whether this has a broader virulence. A comparison vivo to vitro parasite gene demonstrated that 39% regulated transcripts contained URE3 motif recognized...
ABSTRACT There has been renewed interest in the use of flow cytometry for single particle phenotypic analysis particles nanometer size-range such as viruses, organelles, bacteria and extracellular vesicles (EVs). However, many these are smaller than 200 nm diameter, which places them at limit detection commercial cytometers. The reference fluorescence, light-scattering properties akin to those small biological being studied is therefore imperative accurate reproducible data acquisition...