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A discussion on how the best leverage genomic cytometry technologies and methodologies to generate high quality data.

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Robert Salomon, MSc, Operations and Technology Manager, Children's Cancer Institute & Institute for Biomedical Materials and Devices (UTS), Emeritus ISAC SRL Emerging Leader
Luciano Martelotto, PhD, Scientific Director Single Cell Core, Harvard University Medical School

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Important aspects for making flow and mass cytometry reproducible over time and across sites will be reviewed, along with results from multi-site studies.

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Holden Maecker, PhD, Professor (Research), Microbiology & Immunology, Stanford University

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Education and training does not stop at users and staff, outreach at all levels is extremely important. The various ways of implementing both are presented here.

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Alexis Conway, PhD, Flow Cytometry Specialist, Roswell Park Comprehensive Cancer Center; Graduate Course Coordinator, University of Buffalo; ISAC SRL Emerging Leader
Derek Davies, BSc, National STP Training Lead, The Francis Crick Institute, London

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This tutorial will demonstrate workflows of currently available calibration tools for small particle analysis.

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Joshua Welsh, PhD, Postdoctoral Fellow, Translational Nanobiology Section, Laboratory of Pathology, CCR, NCI, NIH; ISAC Marylou Ingram Scholar
John Nolan, PhD, Professor, Scintillon Institute
Edwin van der Pol, PhD, Assistant Professor, Biomedical Engineering & Physics, Amsterdam University Medical Center

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An overview of how label-free, quantitative phase imaging works and its many bio-applications.

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Peter O'Toole, PhD, Director, Bioscience Technology Facility, Department of Biology, University of York

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We will show how the latest computational approaches for the analysis of clinical cytometry data have lowered previous barriers including ease of use and performance.

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Cherie Green, PhD, Senior Scientific Manager, Genentech
Ryan Brinkman, PhD, Professor of Medical Genetics, University of British Columbia
Raphael Gottardo, PhD, Full Professor, Vaccine and Infectious Disease Division at Fred Hutchinson Cancer Research Center; Affiliate Professor of Statistics, University of Washington
Shadi Eshghi, PhD, Scientist, Dept. of Biomarker Development, Genentech
Darya Orlova, PhD, Scientis, Dept. of Cancer Immunology, Genentech

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We will assemble the Make Your Own Cytometer from individual parts to a fully functional flow cytometer.

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Bill Telford, PhD, Senior Associate Scientist, NCI Flow Cytometry Core Laboratory, NCI-NIH

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Discussion of photodetection in flow cytometry and how it can affect scatter plots.

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Slawomir Piatek, PhD, Sr. University Lecturer, New Jersey Institute of Technology; Scientific Consultant, Hamamatsu Corporation

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Enumeration and phenotyping of malignant populations in multiple myeloma patients using multicolor flow cytometry
Bevan Gang, PhD, Principal Scientist, Caprion Biosciences, Inc.

B cell activation and differentiation employ mTORC1 rather than Xbp1 to initiate distinct segments of the unfolded protein response before antibody secretion
Brian Gaudette, PhD, Postdoctoral Researcher, University of Pennsylvania

Mass Cytometry Quality Control Using Heavy Metal-Labeled Reference Spike-In Control Cells
El-ad David Amir, PhD, Chief Executive Officer, Astrolabe Diagnostics, Inc.

A consensus COVID-19 immune signature combines immune protection with discrete sepsis-like traits associated with poor prognosis
Irene del Molino del Barrio, PhD, Senior Technician, University College London

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Boosting the Power of Monitoring the Immunomodulation of Investigational Drugs in Oncology Clinical Trials by Mass Cytometry
Michael Abadier, PhD, Senior Scientist, Takeda Pharmaceutical Co.

Reduction in Non-classical Monocytes Suggests Recruitment to CNS after Acute Traumatic Brain Injury
Monisha Kumar, MD, Associate Professor, University of Pennsylvania

The Functional Landscape of Systemic Immunity in Classical Hodgkin Lymphoma: Traits Associated with Disease, Relapse, and Treatment Response
Pratip Chattopadhyay, PhD, Director, Precision Immunology Laboratory, Associate Professor (Pathology), NYU-Langone Hospital

Integrated trajectories of the maternal metabolome, proteome, and immunome predict labor onset
Xiaoyuan Han, PhD, Postdoc, University of the Pacific

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iIACS2.0: intelligent image-activated cell sorting upgraded
Akihiro Isozaki, PhD, Research Assistant Professor, University of Tokyo

Sequentially addressable dielectrophoretic array for high-throughput large-droplet sorting
Akihiro Isozaki, PhD, Research Assistant Professor, University of Tokyo

Rapid Cluster Cell Sorting Using Rapid Indexing and HyperFinder on the S6 Cell Sorter and the Comparison to the Aria III Cell Sorter
Stephen Perfetto, MA/MS, Chief of the VRC Flow Cytometry Core Facility, NIAID

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An Interpretable Machine Learning Method Enables Biomarker Discovery Through Large-Scale Single Cell Data Integration
Greg Finak, PhD, Senior Staff Scientist, Fred Hutchinson Cancer Research Center

TAILOR:Targeting heavy tails in flow cytometry data with fast mixture modeling
Matei Ionita, BA/BS, PhD Student, University of Pennsylvania

Automated Panel Design with Maximized Sensitivity by Accounting for Spillover Spreading
Sofie Van Gassen, PhD, Postdoctoral Fellow, VIB/Ghent University

Developing scalable integrated single-cell analysis approaches in R using 'Spectre' to build and utilise a multi-disease time-series murine inflammatory single-cell atlas
Thomas Ashhurst, PhD, Specialist, University of Sydney

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Comparison of high dimensional conventional, spectral, and mass cytometry
Diana Shinko, PhD, Mass Cytometry Specialist, University of Sydney

Quantification of the light scattering detection efficiency, background and resolution limit to standardize flow cytometry measurements of extracellular vesicles
Edwin van der Pol, PhD, Assistant Professor, Amsterdam University Medical Center

A Case for Developing a Better Understanding of the Role that Metrology Should Play in Cytometry
James Wood, PhD, Manager, Flow Cytometry Shared Resource, Comprehensive Cancer Center, Wake Forest Baptist Hospital

Multicolor Raman imaging flow cytometry
Nao Nitta, PhD, President, CYBO

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Phenotypic diversity of primary human macrophages interacting with tissue-engineered blood vessels
Beatriz Hernaez-Estrada, BA/BS, PhD Student, Drexel University

Isolation of Chlamydia Trachomatis-specific Human Monoclonal Antibodies Using a Novel Flow Assay
Christopher Pinder, BA/BS, Doctoral Student, Imperial College London

"Immuno-flowFISH”: Next-generation Cytogenetic Assessment provides unique insight into Chronic Lymphocytic Leukemia
Kathryn Fuller, PhD, Associate Professor of Translational Oncology, University of Western Australia

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Defining the phenotype and function of antigen-presenting cells and T cells in the human tumor microenvironment using multi-omic single cell techniques
Florian Mair, PhD, Research Associate, Fred Hutchinson Cancer Research Center

FastMix: A Deconvolution and Differential Expression Analysis Pipeline that Integrates Transcriptomics, Flow Cytometry, and Clinical Covariates
Max Qian, PhD, Associate Professor, J. Craig Venter Institute

Leveraging the power of high parameter cell sorting and single-cell multi-omics to profile intratumoral immune cells in a model of B cell lymphoma
Xiashan Shi, PhD, Senior Scientist, BD Biosciences

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Quantitative evaluation of fluorochrome combinations supports highly multiparametric spectral flow cytometry
Huimin Gu, MA/MS, Systems Engineer, CYTEK Biosciences

Cutting out the Middleman: Leveraging clustering tools’ power to circumvent compensation and facilitate complex sorting and analysis
Jack Panopoulos, PhD, Application Scientist, FlowJo/BD Biosciences

Signed, sealed, delivered, its fluors: Pushing the Envelope in Single Cell Inquiry to the Theoretical Maximum in High Parameter Panel Design
Seddon Thomas, PhD, Research Application Scientist, Phitonix, Inc.

High-throughput, high-dimensional flow cytometry pipeline for large human immunophenotyping studies
Thomas Liechti, PhD, Postdoctoral Researcher, National Institutes of Health

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Dynamic three-photon imaging of lymphoid mouse organs in vivo
Asylkhan Rakhymzhan, PhD, Postdoc, DRFZ Berlin

Fluorescence lifetime light sheet microscopy
Jakub Nedbal, PhD, Postdoc, King's College London

Microfluidic cell sorter with spark-generated cavitation bubble
Jingjing Zhao, PhD, Postdoc, Stanford University

Surface Acoustic Wave Microfluidic Device for On-Demand Droplet Generation, Selective Encapsulation, Cell Lysis, and Pico-Injection
Kirk Mutafopulos, PhD, Senior Scientist, Cytonome/ST; Harvard University

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Standardization of concentration measurements of extracellular vesicles by flow cytometry for medical diagnoses
Edwin van der Pol, PhD, Assistant Professor, Amsterdam University Medical Center

Development of a Flow Cytometry-Based Assay for Clinical Detection and Molecular Characterization of Breast Cancer Disseminated Tumor Cells in Bone Marrow
Elizabeth Chislock, PhD, Senior Research Investigator, Perelman School of Medicine, University of Pennsylvania

High Parameter Flow Cytometry of Pancreatic and Liver Cancer Biopsies
Pratip Chattopadhyay, PhD, Director, Precision Immunology Laboratory, Associate Professor (Pathology), NYU-Langone Hospital

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Extracellular vesicles (EVs) are membrane-derived structures that include exosomes, microvesicles, and apoptotic bodies. In recent years, the importance of EVs has become apparent, as they are key mediators of intercellular communication. However, quantifying and characterizing EVs in a reproducible and reliable manner is challenging due to their small size—exosomes range from 30 to 100 nm in diameter. In this tutorial, we first describe the use of High Gain mode on the Amnis® ImageStream®X Mk II Imaging Flow Cytometer to address the challenges of measuring small particles. In this new High Gain mode, the charge-coupled device (CCD)-camera is adjusted to higher gain settings, increasing the signal obtained from the EV. Object thresholds and masking have also been adjusted to better identify and detect small particles. Preliminary results using murine leukemia virus-sfGFP reference particles have shown up to a 5-fold increase in the number of GFP-positive objects collected in High Gain mode when compared to standard gain on the ImageStream® System. In this study, we demonstrate improved small particle detection, including EVs, using this new High Gain mode. The second part of this tutorial describes the use of a quantitative and specific Vesicle Flow Cytometry assay (vFCTM, Cellarcus Biosciences, Inc.) that takes advantage of the high sensitivity of the Amnis® CellStream® Flow Cytometer. The vFCTM assay consists of a full suite of reagents needed to analyze EVs, including stains, calibrators, and standards, as well as detailed sample preparation and analysis protocols. This assay ensures quantitative and reproducible results. Additionally, we describe the measurement of EV numbers, size, and surface cargo, including tetraspanin and cell-specific markers using the Amnis® CellStream®.

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Amnis® ImageStream®X Mk II High Gain Mode for Increased Small Particle Detection and EV Analysis on the Amnis® CellStream® Using the vFC™ Assay from Cellarcus Biosciences, Inc.
Haley Pugsley, PhD, Manager, Senior Scientist at Luminex Corporation
John Nolan, PhD, CEO, Cellarcus Biosciences, Inc.

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The ImageStream® Mk II and FlowSight® imaging flow cytometers deliver objective, quantitative, and statistically significant high-content data. The richness of the image data and the associated features in the IDEAS® Software (SW) provides huge benefits for researchers, but also introduces complexity in data analysis. We address this issue by providing a new software package that leverages image data with sophisticated, deep learning algorithms to provide solutions to complex problems. The SW provides tools for model training with AI-assisted tagging to facilitate the generation of training data sets. It also creates new AI classifiers that can then be used to classify populations of cells in subsequent data sets. With a database architecture optimized to handle large data sets, an interactive image gallery, and comprehensive report generation, the SW provides an intuitive experience for users, enabling a streamlined data analysis workflow. In this workshop, Dr. Vidya Venkatachalam will explain the features of Amnis AI software—including how it simplifies high dimensional image data analysis—and demonstrate its use on cell data that is difficult to analyze using traditional methods.

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Introducing Amnis® AI – An Image Analysis Software Package Powered by Deep Learning
Vidya Venkatachalam, PhD, Senior Director, Software and Algorithms R&D, Luminex Corporation

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Cell sorting has traditionally been a bottleneck for scientists in the process of completing large workflows. Researchers who require quick and simple cell sorting steps have been impacted by coordination and scheduling difficulties, technological complexity and poor output of high-pressure systems. These challenges all contribute to setbacks in multiple application workflows, both logistically and biologically. NanoCellect Biomedical will present recent examples of solutions to these problems in the field of genomics, cell line development and basic research highlighting the sort of nuclei and fragile cells. Integration of the WOLF Cell Sorting platform into various workflows will re-frame the researcher mindset on how simple, gentle and foolproof cell sorting can truly be.

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Simple, Gentle and Disposable Microfluidic Cell Sorting at Your Bench
Emily Bozek, MSc, NanoCellect Biomedical

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Despite its critical role for the production and release testing of cell products in modern cell therapies, flow cytometry remains a notoriously variable technique. However, reproducibility and consistency are of key concern when monitoring cell production processes and releases of products, such as CAR T cell therapies. Therefore, it is essential that in-process control and quality control (IPC/QC) using flow cytometry fulfill essential standardization criteria. During this pre-recorded tutorial, you will learn about:

• antibody-related reproducibility crisis and implications for antibody-based quality control in cell manufacturing
• advances in recombinant antibody technology and opportunities for flow cytometry assays
• introduction to REAfinity™ Recombinant Antibodies and exemplary data in cell manufacturing assays

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Flow cytometry in cell manufacturing; Reproducible flow cytometry data with lot-to-lot consistent recombinant antibodies
Kalpana Singh, PhD, Product Manager, Flow Cytometry Reagents, Miltenyi Biotech

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Single-cell genomics and other next generation sequencing applications depend strongly on adequate upstream sample preparation. However, harsh conditions during those workflows can easily affect your results in a variety of ways. This is especially troublesome if you are interested in genes related to cellular stress, activation, or metabolism. How can you guarantee you’re using only non-stressed, viable target cells in high purities in your genomics experiments? During this pre-recorded tutorial, you will learn about:

• typical mistakes in sample preparation prior to NGS analysis
• a better way to design your cell sorting steps
• how other scientists have improved their results

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Realize Better NGS Results through gentle cell sorting: The MACSQuant® Tyto® Cell Sorter
Felix Eppler, PhD, Global Product Manager, Miltenyi Biotech

Profiling Cellular Diversity of the Mouse Mammary Epithelium using Single Cell RNA Sequencing Technologies
Quy Nguyen, Kai Kessenbrock Labs

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Scientific research depends upon unhindered access to research technologies. This statement was true before COVID-19 and remains so. The public health crisis has brought many research organisations around the world to rethink access to their shared scientific core facilities. The need to ensure safe democratisation of access to technology resources has erased old habits of paper and pen reservation, and compelled digital management solutions. Since 2011 Stratocore has developed PPMS as a definitive cloud based digital resource management solution providing access to scientific core facilities and shared resources for thousands of researchers all around the world. Supporting and delivering globally our services to the scientific community, we will present an overview of how the PPMS software platform can respond to all your scientific resource management needs across the board, now and far into the future.

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Challenging Times: ensuring ergonomic organization in scientific research facilities with Stratocore
Leonor Heleno Wielgosz, Global Director of Client Relations and Sales, Stratocore

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Bio-Rad are proud to announce the launch of our new proprietary StarBright fluorophores for flow cytometry. These polymer based fluorophores have been designed to be bright and stable, with narrow excitation and emission profiles, reducing the spillover and spread which can be an issue when building large multiplex panels. Find out how they work in most buffers without a reduction in performance with high lot to lot reproducibility.

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StarBrights: Bright Reagents for Bright Ideas
Michael Blundell, PhD, Field Marketing Specialist, Life Science Group, Bio-Rad Laboratories, Inc.

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This tutorial will cover:

• Why use FluoroFinder?
• How to get setup on the platform
• How to design and experiment on FluoroFinder
• New features that are in development!

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Panel Design with FluoroFinder - New Features for CYTO 2020
Jeff Jensen, Director of Sales, FluoroFinder

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Contemporary single cell biology experiments typically rely on isolating single cells by flow cytometry, encapsulation in a droplet, or micro-manipulation. These methods limit the ability to maintain viability, actively select individual cells of interest, and discern more meaningful information from each cell. The CellRaft AIR™ System addresses these limitations and enables streamlined workflows in CRISPR gene editing, stem cell biology, 3D cell culture, and single cell multi-modal analysis. The AIR System is a bench-top instrument capable of imaging, sorting, and isolating single cells or clonal colonies for downstream molecular analysis. The system is based on the CellRaft technology and employs CytoSort Arrays - a proprietary cell culture dish designed with an array of embedded, releasable microwells - to culture, propagate, and phenotype cells prior to isolation. These consumable arrays closely replicate standard in vitro conditions, rendering the array an ideal substrate for time-course phenotyping, drug sensitivity assays, and other imaging-based evaluation prior to isolation for expansion or analysis. The system has been shown to be effective in verifying clonality and ensuring doublets/dead cells aren’t carried downstream. For 3D cell culture applications, growth of organoids in Matrigel and subsequent isolation after weeks of culture has been demonstrated on the system. Workflows in cell line development, antibody discovery, and iPSC reprogramming and differentiation are all well-suited to this approach. In this talk, we will introduce the CellRaft technology, discuss how it compares to traditional methods, and show example data for several applications.

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Functional Cell Sorting: Automated Imaging, Sorting, and Isolation of Single Cells, Clonal Colonies, and Organoids
Jessica, Hartman, PhD, Director of Product Applications, Cell Microsystems, Inc.

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Flow cytometry is a widely applied approach to exploratory immune profiling and biomarker discovery in healthy or pathological conditions. Flow cytometry uses antibodies tagged with fluorochromes or fluorescent dyes for which novel methods allow up to 50 detection parameters. More recently, an innovative technology using stable metal isotopes instead of fluorochromes has emerged. Mass and spectral cytometry has enabled high dimensional analysis and resulted in new research opportunities and challenges. Scientists can no longer manage with the classical bi-axial gating strategy and they need new algorithms, statistical knowledge, terminology, and tools to develop standard analysis approaches. Tercen tutorial material demonstrates how to use workflows, dimension reduction (e.g. PCA, tSNE, UMAP), clustering (e.g. FlowSOM), and marker metrics (e.g. MEM scores). In addition, Tercen gives users the opportunity to integrate different types of data to high dimensional flow cytometry and boost biomarker discovery. Watch our tutorial and join us on Tercen to learn more about High dimensional Flow Cytometry.

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Consideration in High Dimension Flow Cytometry Data Analysis
Faris Naji, CEO, Tercen

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Single cell isolation techniques often suffer from poor efficiency, low recovery, and incompatibility with automation among other limitations. In this presentation, Cellenion will present cellenONE®, a piezo-acoustic dispensing platform with active cell selection for dispensing single cells, from bacteria to cardiomyocytes, into any consumable. Highlighted case studies will demonstrate outstanding accuracy and high viability for cell line development as well as rare cell, single nuclei and whole-genome sequencing.

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cellenONE®: Addressing Limitations in Single-Cell Applications with Bold Innovation
Aileen Murphy, Field Application Engineer, Cellenion