“The BD FACSMelody cell sorter combines quality results with true ease-of-use”, 15 minute product talk
Lisa Bellemare, Technical Applications Specialist, BD Biosciences
The BD FACSMelody cell sorter has been designed to provide quality results with true ease-of-use. The sorting technology incorporated in the BD FACSMelody is built on the BD FACSAria cell sorter platform. The BD FACSMelody combines this proven sorting performance with a new software interface to provide simple and quick cell sorting. This seminar will provide details on our newest cell sorter capabilities, which including a new software, BD FACSChorus, which provides an easy and fast workflow to perform cell sorting for all types of research applications.
Lisa Bellemare obtained a B Sc. from McGill University in Montreal, Canada and a M Sc. From Universite de Montreal, Montreal, Canada. After her graduation, she went on to be a research assistant at UC Davis where she continued her work in cell sorting and mouse models of autoimmune diseases. She has been at BD for four years now, working on everything from cancer models to biofuels, always using her favorite tool, flow cytometry.
“Duraclone cocktails: Standardize and streamline your flow cytometry workflow”, 15 minute product talk
Si-Han Hai, PhD, Supervisor – Flow Cytometry Applications West, Beckman Coulter
Duraclone is a premixed, dried reagent cocktail that can standardize and streamline flow cytometry workflows for clinical research studies. From immunophenotyping to rare event detection, Duraclone reduces variability caused by liquid reagents and multiple users. Originally developed for the multi-site ONE Study of cell therapy for solid organ transplantation, these panels have been optimized through a collaboration between clinical researchers and Beckman Coulter.
Si-Han is the western supervisor for flow cytometry applications based in San Francisco. She supports clinical and research analyzers for Beckman Coulter flow cytometry. She received her PhD in immunology from UCSF and her BS in biology from Brown University and has worked as a research associate at Novartis and Alnylam Pharmaceuticals.
“True-Stain Monocyt BlockerTM, an effective blocking buffer to eliminate non-specific cyanine-like dye-mediated monocyte binding”, 15 minute product talk
Jennifer Boyle, Product Specialist II, BioLegend
Some tandem fluorophores commonly used in flow cytometry have demonstrated a propensity to bind non-specifically to monocytes and macrophages when labeling is done on live cells. Although the effect is not limited to acceptor fluorophores from the cyanine dye family, they are the biggest culprit, and this non-specific binding can limit the ability to do multicolor flow cytometric analysis on lower density antigens. Here, we introduce True-Stain Monocyte Blocker™, which eliminates the nonspecific binding of dyes including PE/Cy7, PE/Cy5, PerCP/Cy5.5,APC/Cy7, APC/Fire™ 750, and PE/Dazzle™ 594. It does not affect antibody binding of monocyte antigens like CD64, CD14, and other tested markers. Multiple fluorophores can be blocked at once, and stability data shows no decrease in signal intensity compared to the conjugate alone. In addition the reagent has no impact on cell viability.
Mock sorting experiments were performed with PBMCs to determine if the True-Stain Monocyte Blocker™ shows inhibitory or enhanced function on inflammatory cytokine/chemokine production. Although more data points are needed to ensure
statistical relevance, our data indicates minimal to no effect above that of the conjugated antibody for IL-6, IL-8, IL-10 and MCP-1 production. We also tested True-Stain Monocyte Blocker™ for its ability to affect PBMC proliferation in response to anti-CD3/anti-CD28 stimulation. We observed minimal to no effect on proliferation by measuring BrdU incorporation.
“High-throughput cytometry using the ZE5 for characterization of immune subsets during development of arthritis in mice”, 25 minute end user talk
Mattias Svensson, PhD, Postdoctoral Scholar, Department of Medicine, UCSD
The use of flow cytometry has been essential for the characterization of distinct immune cells subsets in mice and humans. With the identification of new and rare subsets of immune cells the need for multi-parameter flow cytometry have increased. Furthermore, limitations in sample size (i.e. synovial tissue from joints) has also increased the need for multi-parameter analysis for simultaneous detection of various immune cells. We use the ZE5 flow cytometer for design and rapid evaluation of multi-parameter panels that are used for high-throughput characterization of immune subsets in lymphoid and non-lymphoid tissues of mice. Using this system we have been able to identify and characterize changes within several subsets of immune cells and evaluate their pathological role during development of autoimmune arthritis in mice.
“Expanding your Flow with novel technologies”, 15 minute product talk
Elizabeth Zhang, PhD, Field Application Scientist, ThermoFisher
Flow cytometry is already a powerful technology but innovative solutions are often required to perform more difficult experiments and eliminate experimental constraints. In this brief seminar today, we will share some example data and novel solutions for expanding what you can do using flow cytometry; ranging from using novel sample types, examining rare subsets, overcoming reagent limitations, to pushing the boundaries of particle size.
Dr. Zhang earned her PhD in Immunology at the University of Kansas Medical Center, where she received training in flow cytometry by her advisor, Dr. Thomas Yankee, the former associate professor and scientific director of the flow core. Following her postdoctoral training at La Jolla Institute for Allergy and Immunology, Elizabeth joined Genoptix as a research scientist and was responsible for assay development and validation for flow cytometry clinical trials. Since 2016, she has been working at Thermo Fisher Scientific as a Field Application Scientist for flow cytometry.
“Can your analysis do that? New tools for tomorrow’s problems”, 15 minute product talk
Jack Panopoulos, PhD, Application Scientist, FlowJo
Flow cytometry is reshaping what we know about cell phenotypes, development, behavior and communication. These diverse lines of inquiry share several common analysis problems and pose unique challenges to new investigators as well as the well-entrenched. Thanks to the collaborative effort of many scientists around the world and the software engineers at FlowJo, there are a myriad of tools at your disposal to remedy these challenges; from automated removal of anomalies and calibration to cutting edge clustering and machine learning algorithms.
Jack received his Ph. D. from the University of Southern California from the department of molecular microbiology and immunology in 2006. As a post doc at the Sanford-Burnham Medical Research Institute in La Jolla his work focused on cell cycle dynamics and mechanisms leading to mitotic catastrophe and endoreduplication. He joined FlowJo in 2013, which became part of BD in late 2017. He enjoys developing tools for data analysis and discovery.