Presenters

• Daniel Chiu - University of Washington
  Digital Flow Cytometry (dFC) 

Abstract: We have developed a multi-parametric high-throughput and high-sensitivity flow-based method for counting single molecules, and applied this method to the analysis of individual extracellular vesicles and particles (EVPs). EVPs are promising biomarkers but they are highly heterogeneous and comprise a diverse set of surface proteins as well as intra-vesicular cargoes. Yet, current approaches to the study of EVPs lack the necessary sensitivity and precision to fully characterize and understand the make-up and the distribution of various EVP subpopulations that may be present. Digital flow cytometry (dFC) provides single-fluorophore sensitivity and enables multiparameter characterization of EVPs, including sin­gle-EVP phenotyping, the absolute quantitation of EVP concentrations, and biomarker copy numbers. dFC has a broad range of applications, from analysis of single EVPs such as exosomes or RNA-binding proteins to the characterization of therapeutic lipid nano­particles, viruses, and proteins. dFC also provides absolute quantita­tion of non-EVP samples such as for the quality control of antibodies (Ab), including the concentration of individual and aggregated Ab-dye conjugates and the Ab-to- dye ratio. 

• Lisa Dornish - Allen Institute
  Evaluating the Heterogeneity of Human Peripheral B Cells Using High Dimensional Spectral Flow Cytometry

Abstract: B cells are adaptive immune cells that support durable and effective protection against a wide range of pathogens via antigen presentation as well as differentiation into memory B cells and antibody-secreting plasma cells. Understanding the composition of the human B cell compartment is necessary to effectively isolate and analyze B cells subsets in immune monitoring and systems immunology studies. Here, we present a guide to comprehensively phenotype human B cell subsets using high dimensional spectral flow cytometry. In this guide, we outline the heterogenous B cell identities detectable in peripheral blood and describe their unique surface phenotypes and cellular functions. We further provide guidance for cell surface molecule staining and cytometry data acquisition, informed by factors such as antigenic density, post-translational modifications, cellular activation state and staining conditions. Finally, we offer subset gating strategies and antibody panels that supported our recent analyses of B cells in the context of healthy human aging and pre-clinical Rheumatoid Arthritis. 

• TBD

• Vendor Lightning talks

  • • BD Biosciences - 

    • Cytek Biosciences - 

    • FlowJo - Timothy Crawford

FlowJo™ Version 11:  The next generation of cytometry data analyses solutions. An introduction and orientation. 

   FlowJo™ version 11 is the latest installment of cytometry data analysis tools from the BDB Informatics group (Formerly FlowJo). With a single immersive workbench window, built in quality control, dimensionality reduction and clustering algorithms, plus a new graph gallery and integrated charting capabilities, FlowJo v11 aims to simplify, enable and enhance your interrogation of traditional and next-generation imaging-enabled cytometry data. Please join us for an introduction to the FlowJo v11 platform. 

• • • Standard Biotools - 

    • Ozzette - Arpan Neupane 

• Career Panel

  • • Julie Hill

    • Greg Valtri

    • Garrett Collett

    • TBD

• Perspectives in Data Analysis

  • • Andrew Konecny - Fred Hutchinson Cancer Center

    • Hugh MacMillan - Fred Hutchinson Cancer Center

    • Ana Kus - Benaroya Research Institute

    • TBD 

• Keynote

  • Evan Newell - Fred Hutchinson Cancer Center

Improving coordinated immune profiling through collaborative innovation 

• Beckman Coulter Happy Hour Talk

  • Rafael Villasmil - National Eye Institute 

Integrating Nano-Cytometry: A Core Lab’s Perspective on the CytoFlex Nano