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  • Contains 3 Component(s), Includes Credits Includes a Live Web Event on 09/28/2022 at 12:00 PM (EDT)

    A CYTO U Webinar presented by Andrew Filby, PhD Keywords: Shared Resource Laboratories, team science, health research culture, equality and diversity

    About the Speaker

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    Andrew Filby, PhD
    Director of the Newcastle University Flow Cytometry and Single Cell Technologies Shared Resource Laboratory
    New Castle University

    Dr. Filby is currently the director of the Newcastle University Flow Cytometry and Single Cell Technologies Shared Resource Laboratory. He also leads the “Innovation, Methodology and Application” (IMA) crosscutting research Theme in the Faculty of Medical Sciences (FMS), where he oversees about 350 members from different technology and methodology backgrounds that includes both academic and technical job families. Dr. Filby is also a director of the Newcastle University Centre of Research Excellence (NUCoRE) in Biomedical Engineering. Dr. Filby and his team have an active program of research that includes the development of novel methods for single cell analysis and evaluating new technologies. He publishes in high-impact journals such as Science and Nature and sits on several funding panels for equipment grants. He recently won the Times Higher Education (THE) “Outstanding Technician of the Year” in 2021 for his work on breaking down barriers and establishing parity of esteem for non-academic researchers.

    Webinar Summary

    In this webinar Dr. Filby will discuss how the “Shared Resource Laboratory” (SRL) can be a catalyst for establishing a culture of team science, inclusivity, and health research culture. He will also discuss his role as lead of the “Innovation, Methodology and Application” (IM) cross-cutting research theme within the faculty of medical sciences at Newcastle University. Dr. Filby will also address the imbalance that sometimes exists between academic and technical job families and how he has worked via the “Innovation, Methodology and Innovation” (IMA) research Theme that he leads to try and overcome this.

    Learning Objectives

    • How the shared resource laboratory (SRL) can be a catalyst and “rally point” for team science and a healthy research culture.
    • How the right attitudes to research culture can create an inclusive and effective environment for success.
    • How technology and methodology expertise can drive discovery and impact.

    Who Should Attend

    • Anyone working in or interested in working in an SRL.
    • Anyone interested in or involved in team science.

    CMLE Credit: 1.0

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  • Contains 3 Component(s), Includes Credits

    A CYTO U Webinar presented Kewal Asosingh, PhD Keywords: Endothelial, flow cytometry, rare event detection

    About the Speaker

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    Kewal Asosingh, PhD
    Scientific Director
    Cleveland Clinic Lerner Research Institute

    Dr. Asosingh is a principal investigator and scientific director for Flow Cytometry at the Cleveland Clinic Lerner Research Institute. He is a past ISAC Scholar, Cytometry Part A Associate Editor, ISAC Flow Cytometry Content, and Education Committee member. His flow cytometric research is centered on lung vascular biology.

    Webinar Summary

    Endothelial cells are cells that reside in tissues and circulate in the bloodstream in small numbers. Immunophenotyping these cells has several challenges, like how to detect endothelial cells, how to gate them, and how to figure out if they're really endothelial. All of this will be covered in this webinar.

    Learning Objectives

    • Immunophenotype of murine and human circulating and organ-specific endothelial cells
    • Endothelial cell gating strategies and common pitfalls
    • The dos and don'ts of functional endothelial cell characterization

    Who Should Attend

    Flow cytometry SRL members and anyone interested in endothelial cell flow cytometry.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits

    A CYTO U Webinar presented by Felix J. Hartmann, PhD Keywords: Cellular metabolism, tumor immunology, multiplexed imaging

    Want this webinar for free? Upgrade to Gold or Platinum membership here! https://isac-net.org/page/Memb...

    Preview of Webinar

    About the Speaker

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    Felix J. Hartmann, PhD
    Helmholtz Young Investigator
    German Cancer Research Center (DKFZ), Heidelberg, Germany

    Dr. Hartmann received a BSc and MSc in molecular biotechnology from the University of Heidelberg, Germany and his PhD from the University of Zurich, Switzerland for his research on T cell effector functions in human autoimmune diseases. In 2017, he joined Stanford University as a postdoctoral fellow to study anti-tumor immunity and its metabolic regulation. Since 2021, Dr. Hartmann is an independent group leader at the German Cancer Research Center (DKFZ), Heidelberg, Germany. His research combines single-cell and imaging proteomic technologies with novel biological assays to reveal interactions of immune cells with their local environment and how these interactions impact clinical outcome in human cancer. Most recently, he has developed a novel approach that enables analysis of cellular metabolism in individual cells and with spatial resolution.

    Webinar Summary

    Cellular metabolism regulates immune cell activation, differentiation and effector functions, but current metabolic approaches lack single-cell resolution and simultaneous characterization of cellular phenotype. In this webinar you will learn about a novel approach to characterize the metabolic regulome of single cells together with their phenotypic identity. The method, termed single-cell metabolic regulome profiling (scMEP), quantifies proteins that regulate metabolic pathway activity using high-dimensional antibody-based technologies. You will learn how mass cytometry (cytometry by time of flight, CyTOF) was employed to benchmark scMEP against bulk metabolic assays by reconstructing the metabolic remodeling of in vitro-activated naive and memory CD8+ T cells. You will see the approach being applied to clinical samples to identify tissue-restricted, metabolically repressed cytotoxic T cells in human colorectal carcinoma. Combining the method with multiplexed ion beam imaging by time of flight (MIBI-TOF), allowed uncovering of the spatial organization of metabolic programs in human tissues, which indicated exclusion of metabolically repressed immune cells from the tumor–immune boundary. Overall, the approach described in this webinar enables robust approximation of metabolic and functional states in individual cells.

    Learning Objectives

    • Understand the concept of metabolic regulome profiling using antibodies and its relationship with cellular metabolism
    • See how metabolic regulome profiling was implemented using high-dimensional mass cytometry (CyTOF) and multiplexed ion beam imaging (MIBI)
    • Realize the potential of this approach for clinical biomarker discovery

    Who Should Attend

    Scientists at all levels interested in cellular metabolism, immunology and cancer research, singe-cell analysis and imaging

    CMLE Credit: 1.0

  • Contains 6 Component(s), Includes Credits

    A CYTO U Webinar presented by Jessica Perea Houston, PhD The event will include a live discussion with the speaker about her contribution in promoting equality and inclusion in the field. Keywords: Women in science, cytometry instrumentation, diversity & inclusion

    Webinar Preview

    About the Speaker

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    Jessica Houston, PhD
    Professor
    New Mexico State University

    Jessica Perea Houston, PhD, is a professor in chemical & materials engineering at New Mexico State University (NMSU) in Las Cruces, NM (2009-present). Jessica received her PhD in chemical engineering from Texas A&M University (2005) and was a director’s postdoctoral fellow at the Los Alamos National Laboratory Bioscience Division (2006-2009). She is an alumna of NMSU (’00) and is from Santa Fe, NM.

    Her research expertise is biomedical instrumentation development with an emphasis on time-resolved flow cytometry systems development, biophotonics, and optofluidics. Jessica directs a flow cytometry instrumentation lab at NMSU, has advised over 60 graduate students, undergraduate students and/or postdocs throughout the 12.5 years she has been at NMSU. Her research is currently supported by a National Institutes of Health R01 grant “Microflow time-resolved flow cytometry for FRET and Fluorescent Protein Development.”

    Jessica has been active in multidisciplinary research projects that have involved many collaborators, institutions, and international partners. She and her team have presented over 400 abstracts at conferences that range from local to international. She has published over 50 papers related to her research, has one patent, and has been invited to over 30 different colloquia and conferences, where she has made formal invited presentations related to her research. Jessica was a Faculty Fulbright Scholar in Japan for 6 months in 2018. She has received numerous grants and awards including her current NIH R01 award and the NSF CAREER award in 2012. She became a Scholar of the International Society for the Advancement of Cytometry in 2012 and won Best Paper in the journal Cytometry Part A in 2015. She has also received many awards at NMSU including the Synergy-One award (NMSU College of Engineering) Outstanding Junior Faculty (NMSU Hispanic Faculty and Staff Caucus); the Early Career Award (NMSU Research Council) and the Distinguished Career Award (NMSU Office of the Vice President for Research)

    Jessica is currently associate editor for the Journal, Cytometry Part A and is a standing member of the Cell & Molecular Technologies Study Section for the National Institutes of Health Center for Scientific Review. Jessica has been actively involved in ISAC for 10 years and a member of ISAC for 16 years. She is ISAC Treasurer and Chair of the Finance Committee. She is also an executive committee member ISAC. She helped found the CYTO Women taskforce within ISAC and has been chair of the Scientific Communications Committee. She was on Council from 2012-2016. Jessica also is/has been a member of the following committees: Scholars, Leadership Development, CYTO Women, Transition & Personnel, Committee Liaison Collective, Mission Continuity, Awards, Nominating, CYTO Planning, and Fundraising. She has continued to participate in the planning of the CYTO congress and has attended most CYTO meetings since 2008. In other organizations, Jessica is a leader—she was chair of the entire Biophotonics session for the Conference on Lasers and Optics (CLEO) for 3 years (2017-2019). In 2021 Jessica was invited to co-chair the Photonics West BiOS Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues conference in San Francisco, CA.

    Jessica also has experience participating, learning, and working in the general area of diversity and inclusion. As a Hispanic female, Jessica has been quite interested in advancing minority initiatives. She is involved in many minority societies and programs, and attends workshops and meetings related to underrepresented minority (URM) priorities. She was a participant in the 2015 cohort of the Linton-Poodry SACNAS Leadership Institute, attended several Society of STEM Women of Color retreats and is a SSWOC member. She has participated in the 2019 AAHHE (American Association of Hispanics in Higher Ed) conference to formulate a strategic plan with the NMSU Vice President for Research. She and her students regularly attend the SACNAS and ABRCMS annual meetings. She participates in many workshops related to diversity in STEM including recent ones held on An Inclusive Academy: Achieving Diversity and Excellence (by authors: Abigail Stewart and Virginia Valian) and a Women in STEM book club. She has advised more than 30 minority undergraduates in research, and seven graduate students and postdocs. In total Jessica has mentored more than 50 undergraduates in research, advised six PhD degrees and eight MS degrees. She is now mentoring two minority RISE PhD students, one minority MARC undergraduate researcher, three minority undergraduate research students, and two postdoctoral fellows.

    Webinar Summary

    In cytometry, quantitative interpretation of cell function and morphology at a single-cell level can be accomplished through many different approaches that range in how the detectors function, excitation light is delivered, cells are moved (fluidics), and data are processed. Moreover, cytometers have been adapted for imaging, mass spectrometry, full spectral analyses, unique sorting/lab-on-a-chip approaches, and machine learning. Yet among the plethora of ways a cell can be quantified as it passes a laser beam, fluorescence decay is rarely captured. The average time fluorescent molecules within the cell spend in the excited state is typically ignored because of the challenges related to evaluating fluorescence lifetimes at a high throughput. Over several years, our laboratory has taken cytometers and introduced a variety of lifetime technologies onto these systems for analysis and sorting. Lifetime sensing with cytometry is achievable using a wide range of cytometric architectures. Some examples of these approaches will be discussed, as will new protocols and applications we are developing. The main cellular assays that we focus on include measurements of Förster resonance energy transfer and metabolic mapping of cells by NAD(P)H lifetime sensing. It is non-trivial to capture heterogeneous time-resolved information from several different excitable molecules when excitation is observed from single cells and particles in fluidic states. Therefore, the opportunities for addressing such challenges will also be presented. Additionally, the experiences had by Dr. Houston during her career will be discussed including current opportunities for diversifying academia and the challenges for women in engineering.


    Learning Objectives

    • Discuss the benefits of time resolved flow cytometry
    • Compare and contrast lifetime analysis for imaging and cell sorting
    • List various ways in which lifetime as a cytometric parameter enhances cytometry assays and protocols 
    • Discuss current challenges faced in academia by women in STEM
    • List opportunities for enhancing diversity at your institution and developing plans for inclusion and equity.

    Who Should Attend

    The broad cytometry community and scientists interested in topics related to diversity and inclusion.

    CMLE Credit: 1.0

    • Register
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      • Silver - Free!
      • Gold - Free!
      • Platinum - Free!
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      • ISAC Staff - Free!
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  • Contains 4 Component(s), Includes Credits Recorded On: 03/02/2022

    A CYTO U Webinar presented by Rafael J. Argüello, PhD Keywords: immunometabolism, functional method, flow cytometry, single cell resolution, SCENITH

    Preview of Webinar


    About the Speaker

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    Rafael J. Argüello, PhD
    Center of Immunology of Marseille Luminy 

    Rafael Argüello studied biology at the University of Buenos Aires and did his PhD in immunology. He is a tenured CNRS researcher and leader of the immuno metabolism and translation team, in the DeCiBEL laboratory at the Centre d'Immunologie de Marseille Luminy (CIML, France). In 2020 he patented and published SCENITH (PCT/EP2020/060486) and became a Marylou Ingram Scholar of the International Society for the Advancement of Cytometry. He has been awarded with grants as PI from Canceropole, Inserm tranfert, ImmunoX UCSF, ECOS-Sud, Centuri and ANR Young Investigator award 2021. He has recent publications in Cell Metabolism, Cell, Nature immunology, Nature Cancer and EMBO Journal and has created a large collaborative network to study immunometabolism ex-vivo. 

    Webinar Summary

    Interest in immunometabolism is growing exponentially due to its implication in infection, autoimmunity, cancer, and immunotherapies. However, studying immunometabolism ex-vivo has been technically challenging for the last decades. In the last two years, efforts from many different labs have generated a toolbox of functional and phenotypic markers that allow to characterize the metabolic profile of immune cells by flow cytometry. In this webinar we will overview, update, and compare the different tools available and guide you to perform your first immunometabolic studies by flow cytometry. 

    Learning Objectives

    In this webinar you will learn the latest advances in single cell technologies to study immunometabolism. (I will NOT be speaking about the Krebs cycle.) You will learn about functional and phenotypic tools to study metabolism by flow cytometry. In particular, you will learn to interpret experiments and results from SCENITH and other methods.

    Who Should Attend 

    Master, PhD, Postdocs, and PIs interested to explore the immunometabolism field by flow cytometry.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 12/08/2021

    A CYTO U Webinar presented by Kylie Price and Dagna Sheerar Keywords: Novel funding sources, philanthropy, relationship building, instrument procurement, infrastructure

    About the Speakers

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    Kylie Price
    Head of Research Technology and Senior Staff Scientist
    Malaghan Institute of Medical Research

    Kylie Price is a senior scientist and the head of research technology at the Malaghan Institute of Medical Research. She has 18 years of experience providing strategic, scientific, and operational direction in multidisciplinary environments. Kylie has strong stakeholder management and engagement skills and has attracted more than $15 million of philanthropic funding over the past twelve years, supporting the creation of a distinguished center of research excellence, the Hugh Green Cytometry Centre (HGCC). Kylie leads a team of eight highly-skilled technology scientists in providing supporting to over 80 scientists at the Malaghan Institute. The HGCC provides access to cutting-edge technology platforms (including flow and spectral cytometry, histology, bioimaging, genomics, and bioinformatics) and advises multiple research groups both nationally and abroad. Kylie also has a strong leadership track-record that includes organizing high-profile networking events, such as CYTO Asia Singapore 2017, and directing international organizations, such as the Australasian Cytometry Society of which she is former president. Kylie is a two-time NZ Woman of Influence Awards finalist and finalist of the 2021 NZ High Tech Awards. She was the first New Zealander elected to the International Society for Advancement of Cytometry Council and was elected ISAC Secretary in 2020.

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    Dagna Sheerar

    Flow Cytometry Technical Director
    University of Wisconsin
    Carbone Cancer Center Flow Cytometry Laboratory

    Dagna Sheerar has been working in flow cytometry shared resource laboratories for over 20 years and is currently the technical director of the Flow Cytometry Laboratory at the University of Wisconsin - Madison Carbone Cancer Center. Her areas of interest are core management and user education. Since becoming technical director, Dagna has successfully added, on average, one instrument per year to the Flow Lab through various grant programs, institutional purchases, or “crowd funding.” Dagna has been involved in the Great Lakes International Imaging and Flow Cytometry Association (GLIIFCA) for almost as long as she has been working in cores, and she recently served as GLIIFCA president from 2020-2021. She is a member of ISAC and serves on the SRL Content Task Force. She serves as host, organizer, and instructor for the Annual Course in Cytometry, and she is currently in the planning stages for the 45th Annual Course to be held at the University of Wisconsin in June of 2022. Dagna has presented at GLIIFCA, the annual CYTO Meeting, and the Association of Biomolecular Resource Facility (ABRF) annual meetings. Dagna also served as the Interim Director of Campus Research Cores at UW – Madison in 2020.  

    Webinar Summary

    There is often a lot of pressure on SRL managers to run a partial- or full-cost-recovery core facility which presents many challenges. Identifying and securing external funding (in any form) can alleviate this pressure. This tutorial will cover various funding strategies for SRLs, looking at diversifying funding streams. Dagna Sheerar will provide an overview of traditional and novel methods for funding the purchase of instrumentation, including various grant programs, institutional purchases, and “crowd-funding.” Kylie Price will look at how to find, maintain, and grow philanthropic relationships as well as how to maximize opportunities with high-net-worth individuals/groups.

    Learning Objectives

    • Identify novel funding sources for shared resource laboratories.
    • Leveraging usage data and research need to build support.
    • How best to present core technologies and core facility services to entice funding.
    • How to engage stakeholders.
    • Provide tips around how to build and grow philanthropic relationships.

    Who Should Attend

    SRL Managers, Flow Core Directors, SRL Staff.


    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits

    A CYTO U Webinar presented by Marta Monteiro, PhD and Julie Auger Keywords: SRL, benchmarking, quality assurance

    About the Speakers

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    Marta Monteiro, PhD
    Head of Flow Cytometry & Antibodies
    Core Facility Speaker
    Instituto Gulbenkian de Ciência

    Marta has a PhD in immunology from the University of Paris. She is head of Flow Cytometry & Antibodies at the Gulbenkian Institute for Science (IGC), in Portugal, and also the core facility speaker, representing all IGC facilities. Before that, Marta worked as a researcher for 15 years in Portugal and abroad and was an assistant professor at he Medical School of University of Lisbon. She also was the head of the R&D department of a multinational pharmaceutical company. She is author of many scientific publications and co-inventor of a patent that was an award-winning entrepreneurship project and the foundation of a start-up. With her strong scientific know-how and practical input in core facility management, Marta also collaborates with Agendo for business development and marketing.

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    Julie Auger
    Executive Director
    Research Core Facilities Program
    University of California Davis

    Julie A. Auger, has been a member of the cytometry community since 1985 and is widely recognized for her expertise in the operation and administration of multi-technology, multidisciplinary academic resource centers. She serves as the executive director of the Research Core Facilities Program at the University of California Davis where she is responsible for development of a central support structure to optimize organization and administration for over 100 shared research facilities. Prior to her current position at UC Davis, Julie developed similar programs at UC San Francisco and The University of Chicago. She developed her core facility management expertise at the University of Illinois Urbana-Champaign and at the University of Chicago, where she developed internationally-reputed flow cytometry core facilities.

    Webinar Summary

    At research institutions, shared resource laboratories (SRLs) managers face increasing expectations to maximize facility usage and optimize operation. Benchmarking is a helpful exercise for organizations to identify their strengths and opportunities for development with the aim of maximizing performance. External assessment of the SRLs can be used as a benchmarking strategy to perform an objective and unbiased analysis of the standpoint of those services, providing valuable recommendations for improvement on different areas. In this webinar, we will present a successful case study of a first external assessment of the scientific core facilities of a research institution. We will provide an overview of the different steps, from the initial engagement of the entire community to prepare the exercise, to the benefits it conveyed and that are seen at present.

    Learning Objectives

    • Usefulness of benchmarking for SRLs.
    • Different steps of an external assessment process of SRLs.
    • Benefits that an external assessment process of SRLs can bring to an entire institution.
    • Definition of Key Performance Indicators/metric for SRLs.

    Who Should Attend

    SRLs managers, SRLs technicians, institutional executive and scientific managers.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits

    A CYTO U Webinar presented by Dr. Kewal Asosingh Keywords: Solid tissue, disaggregation, single-cell suspension

    About the Speaker

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    Dr. Kewal Asosingh
    Scientific Director for Flow Cytometry
    Cleveland Clinic Lerner Research Institute

    Dr. Asosingh is a principal investigator and scientific director for flow cytometry at the Cleveland Clinic Lerner Research Institute. He is a past ISAC Scholar, Cytometry Part A associate editor, ISAC Flow Cytometry Content subcommittee chair, and member of the Education Committee. He has more than 20 years of experience in solid tissue disaggregation into a single-cell suspension.

    Webinar Summary

    A good single-cell prep is essential for any flow cytometry and single-cell omics experiment. This webinar outlines the principles and provides a general guide of steps to consider for solid tissue disaggregation.

    Learning Objectives:

    • Basics of solid tissue composition.
    • What is a “good” single-cell prep in flow cytometry and single-cell RNAseq experiments?
    • The dos and don’ts in designing a tissue disaggregation protocol.
    • Evaluation of the quality of a single cell prep and common pitfalls. 

    Who Should Attend

    Flow cytometry SRL staff and anyone performing flow cytometry using solid tissue as starting material. 

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 10/13/2021

    A CYTO U Webinar presented by Sherry Thornton, PhD and Julia Fernandez-Rodriguez, PhD Keywords: grants, funding, SRL, tips and tricks for funding, funding sources

    About the Speakers

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    Sherry Thornton, PhD
    Professor of Rheumatology
    Director, Research Flow Cytometry Core

    Dr. Sherry Thornton been involved in flow cytometry for over 19 years both in projects as an investigator and in provision of services as a core director. She is a field service professor in the Department of Pediatrics at the University of Cincinnati College of Medicine (UCCOM) in the Division of Rheumatology at Cincinnati Children’s Hospital Medical Center (CCHMC). Her main role is to direct the research flow cytometry core and provide services to over 190 investigators and their labs both at CCHMC and UCCOM. She is highly involved in education and shared facilities nationally and internationally. She currently serves as chair of the Education Committee of the International Society for the Advancement of Cytometry (ISAC) and is a past chair and current member of the Flow Cytometry Research Group for Association for Biomedical Resource Facilities (ABRF). She also was involved in the launching and the continuation of the ABRF mentorship program as a member of the Career Development Committee. In striving to provide education and state-of-the-art flow cytometry for core users, she has been supported by two NIH P30 grant mechanisms for over 15 years and has been awarded two NIH Shared Instrumentation Grants to provide access to user-friendly based cell sorting 24/7 and high parameter flow cytometry. 

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    Julia Fernandez-Rodriguez, PhD
    Sahlgrenska Academy
    University of Gothenburg, Sweden

    Julia's research career in cell and molecular biology and core facility training experience has provided her with an excellent background in multiple life sciences disciplines, as well as in the management, operation, and coordination of an imaging research infrastructure and its training activities at the national and international level. Since 2003 Julia has been responsible of the Centre for Cellular Imaging, an open-access Correlated Multimodal Imaging Facility that provides technical and scientific excellence by integrating multiple imaging technologies with image processing and analysis tools in a single core. Julia's main interest is to provide expertise in correlated multimodal imaging workflows (from experimental design to image acquisition and analysis) tailored to various research domains within the life sciences. In 2016, Julia was awarded one of the 15 Research Infrastructure Fellows grants by the Swedish Foundation for Strategic Research. At the university core facility, Julia is also involved in the education and training of students and researchers through a series of courses, seminars, and workshops, often in collaboration with other universities in Sweden and abroad and with industrial partners. Julia's responsibility is to organize and lead these events and ensure that the scientific community receives the appropriate basic or advanced training on different microscopy methods. Julia's overall aim is to have students and researchers foster a deep understanding of basic and advanced methods used in microscopy to tackle their questions about the most appropriate probes and instruments. Furthermore, she has supervised core internship students in bioimaging, promoting future career possibilities and introducing core facility work as a possible career direction. At the national level, she is the scientific coordinator of the National Microscopy Infrastructure in Sweden and a member of the Boards of the Nordic Microscopy Society (SCANDEM) and the Bridging Nordic Microscopy Infrastructure (NordForks). Julia is also connected to several other European facilities such as the European Light Microscopy Initiative, ELMI (member of the Steering committee since 2008), and the Euro-BioImaging ERIC consortium (member of the Nodes Board). She is also president of the Core Technologies for Life Sciences Association (CTLS). Further, she represent Sweden in the Management Committee and is the coordinator of the Short-Term Scientific Missions of the European COST Action COMULIS (CA17121) -funded network in Correlated Multimodal Imaging in Life Sciences.

    Webinar Summary

    Participants are invited to learn tips and tricks regarding funding for shared resource laboratories (SRL). Speakers will address NIH and European funding sources, focusing on how participants can support their cytometry SRL through instrumentation or other grant mechanisms.

    Learning Objectives

    • Identify funding sources for shared resource laboratories.
    • Determine basic requirements for successful core laboratory grant submissions.
    • Provide specific tips for successful grant submissions.
    • Determine common pitfalls for grant applications.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 10/12/2021

    A CYTO U webinar presented by Florian Mair, PhD and Thomas Liechti, PhD Keywords: Dendritic cells, Monocytes, High-dimensional flow cytometry, Panel design, Unsupervised data analysis

    About the Presenter

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    Thomas Liechti, PhD
    Postdoctoral Researcher
    ImmunoTechnology Section
    Vaccine Research Center
    National Institutes of Health

    Thomas Liechti obtained his PhD in immunology and microbiology at the University of Zurich in 2017 and is currently a postdoctoral researcher in Mario Roederer’s group at the Vaccine Research Center of the National Institutes of Health in Bethesda (USA). His main interest is high-dimensional flow cytometry and human immunology. During his postdoctoral training, he established a 28-color flow cytometry sample processing and analysis pipeline to assess the contribution of genetic and environmental factors to human immune homeostasis in a cohort of more than 3000 individuals.  


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    Florian Mair, PhD
    Research Associate | Cytometry Specialist
    Fred Hutchinson Cancer Research Center

    Florian Mair graduated with a PhD from the University of Zurich, Switzerland, in 2014 and is currently working at the Fred Hutchinson Cancer Research Center in Seattle, USA, as a research associate. During the past decade, he has been involved extensively with different cytometry platforms (conventional, spectral and mass cytometry) as well as scRNA-seq techniques and developed an interest in applying novel analysis approaches for single cell data. He has been actively engaged in teaching flow cytometry courses, including systematic panel design and analysis of high-dimensional cytometry experiments. 

    Webinar Summary

    Human dendritic cells (DCs) and monocytes are critical components of the innate immune system and important for orchestrating adaptive immunity. In recent years high-dimensional single cell technologies, such as flow cytometry and single cell RNA-seq, dissected the heterogeneity of human DCs and monocytes more precisely, suggesting more robust markers to unequivocally define these subsets. Based on these results and our recently published phenotype report (Mair F. and Liechti T. Cytometry Part A, 2021), we will discuss strategies to better define and characterize human DCs and monocytes using a combination of traditional and recently discovered markers. We will show practical examples of how the combination of thorough panel design and unsupervised data analysis can help to dissect heterogenous immune populations. This webinar will be split into three parts:

    • A description about the phenotypic and functional characteristics of human DC and monocyte subsets with a focus on how the emergence of single-cell technologies improved our understanding of the heterogenous landscape of human phagocytes.
    • High-dimensional panel design for characterization of human DCs and monocytes with a more specific focus on marker selection and the incorporation of biological knowledge into this process.
    • An overview of how unsupervised data analysis approaches can improve the delineation of human DC and monocyte subsets.

    Learning Objectives

    • Learn about the phenotypic and functional characteristics of human DCs and monocytes.
    • Understand strategies for marker selection in high-dimensional flow cytometry panel design.
    • Learn how unsupervised data analysis can guide the analysis of human DCs and monocytes.

     Who Should Attend

    Scientist interested in studying human dendritic cells and monocytes using flow cytometry. In addition, flow cytometrists with a general interest in high-dimensional flow cytometry panel design.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 09/21/2021

    A CYTO U Webinar presented by Jakob Zimmerman, PhD Keywords: Intestinal microbiota, Gnotobiotic mice, Bacterial flow cytometry and cell sorting, Host-microbe mutualism, Inflammatory Bowel Disease

    About the Speaker

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    Jakob Zimmerman, PhD
    University of Bern
    Department for BioMedical Research

    Dr. Zimmermann obtained his PhD in the lab of former ISAC president Andreas Radbruch at the German Rheumatism Research Center in Berlin, Germany, working on Th cells in the pathogenesis of IBD, while also establishing new methods for the flow cytometric interrogation of the intestinal microbiota. As a postdoctoral fellow supported by a Marie-Curie fellowship by the European Commission, he moved to the lab of Andrew Macpherson at the University of Bern, Switzerland, to further specialize in host-microbe mutualism. As an ISAC Marylou-Ingram scholar, he is currently leveraging the power of bacterial flow cytometry when combined with the robustness of defined gnotobiotic model microbiotas.

    Webinar Summary

    The intestinal microbiota has been implicated in nearly all aspects of human health, yet our mechanistic understanding of these microbial consortia and their interaction with the host remains superficial. This webinar shall address how flow cytometry and cell sorting of gut bacteria can contribute to deepen this knowledge. It’ll involve key advantages over other techniques, examples for its application as well as practical guidance and pitfalls when doing microbiota flow cytometry.

    Learning Objectives

    Attendants should learn how flow cytometry can contribute to research on the intestinal microbiota and which questions can best be answered using microbiota flow. Topics that will be discussed include how gnotobiotic mice with defined microbiotas can be leveraged for robust microbiome research and why they are particularly powerful when combined with microbiota flow cytometry and cell sorting. A key learning objective is also how to do microbiota flow cytometry and sorting as well as important controls and potential pitfalls.

    Who Should Attend

    As the gut microbiota affects almost all human (and mouse…) organ systems, the webinar is targeted at immunologists and cancer researchers just as much as at microbiologists and microbiota scientists.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 09/08/2021

    A CYTO U Webinar present by Jessica B. Back, PhD, SCYM(ASCP)CM and Ann Marie DesLaurieres-Cox Keywords: Emotional Intelligence, user interactions, interpersonal relationships, social, empathy

    About the Presenter

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    Jessica B. Back, PhD, SCYM(ASCP)CM
    Deputy Director
    Microscopy, Imaging, and Cytometry Resources Core
    Karmanos Cancer Institute, Wayne State University

    Jessica B. Back, Ph.D., SCYM(ASCP)CM is a research scientist and deputy director of the Microscopy, Imaging, and Cytometry Resources (MICR) Core at the Karmanos Cancer Institute and Wayne State University in Detroit, MI. She received her PhD in biochemistry from Wayne State University and completed her post-doctoral training in tumor immunology at the Karmanos Cancer Institute. Jessica is an ISAC councilor, a former ISAC SRL emerging leader (2015-2019), a member of the Great Lakes International Imaging and Cytometry Association (GLIIFCA) Board of Directors, and chair of the American Society of Clinical Pathologists (ASCP) Board of Certification Cytometry Examination Committee. As Deputy Director of an SRL, much of her focus is on efficient and effective delivery of services to facility users. Her research interests focus on the tumor microenvironment, particularly on the role the immune system plays in cancer therapies and tumor regression.

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    Ann Marie DesLaurieres-Cox
    Assistant Manager, Flow Cytometry Core
    University of Michigan

    Ann Marie DesLauriers-Cox is the assistant manager of the Flow Cytometry Core Facility (FCCF) at the University of Michigan in Ann Arbor, Michigan. She received her BS in clinical lab sciences at Eastern Michigan University. Ann Marie has been a technician in the Flow Core for 25 years, where she focuses on customer relations, training, maintenance, and troubleshooting of the Flow Cytometers. She has presented previous “soft skills” workshops at both CYTO and GLIIFCA conferences.

    Webinar Summary

    Shared Resources Laboratory (SRL) personnel often assume a role as scientific mentor to trainees within their institutions. In this position, they may see and hear things from these trainees that fly under the radar of their mentors and occasionally result in an overflow of emotions in the SRL facility. As such, SRL staff need to be equipped with emotional intelligence to navigate these interactions, provide support to users if needed, and alert the mentor or administration if required, all while maintaining a professional working environment. The goal of this CYTO U session is to provide a brief introduction to emotional intelligence and a framework SRL personnel may use to navigate these interactions with users and colleagues.

    Learning Objectives

    • Familiarize the audience with the concept of emotional intelligence.
    • Introduce a framework for the practical application of emotional intelligence within the SRL.

    Who Should Attend

    • RL Managers and Personnel.

     

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits

    A CYTO U webinar presented by Anna Belkina, MD, PhD Keywords: data analysis, algorithms, UMAP, t-SNE, dimensionality reduction

    About the Presenter

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    Anna Belkina, MD, PhD
    Assistant Professor of Pathology and Laboratory Medicine
    Director, Flow Cytometry Core Facility
    Boston University School of Medicine

    Anna C. Belkina is an assistant professor of pathology and laboratory medicine and the director of the Flow Cytometry Core Facility at Boston University School of Medicine. She received her MD degree from Russian State Medical University in Moscow and her PhD degree from Boston University School of Medicine investigating the epigenetic regulation of inflammatory responses driven by bromodomain proteins. Anna’s research is focused on the intersection of immunology and computational biology and her research efforts include investigating the immune landscape of chronic inflammatory diseases and developing computational techniques to assess high-parameter single cell cytometry data. She has designed the opt-SNE algorithm that is now widely used for the visualization of multidimensional cytometry datasets. 
    Anna is an active member of ISAC and has been named a 2015–2019 ISAC SRL Emerging Leader. She is a member of the ISAC Council elected for the 2020–2024 term.

    Webinar Summary

    Visualization of multiparameter datasets is a staple task in the data analysis pipeline. Over last few years, multiple dimensionality reduction algorithms have been adopted for visual presentation of cytometry data to aid identification of novel cell populations and biological trends. In this webinar, we will discuss the basic principles of these approaches and identify benefits and drawbacks of several mainstream algorithms including popular variants of t-SNE and UMAP. We will specifically highlight the importance of hyperparameter optimization for cytometry datasets and practical considerations of choosing the suitable computational environment for your analysis. 

    Learning Objectives

    • Familiarize the audience with recent advances in dimensionality reduction approaches in cytometry datasets. 
    • Introduce and discuss the basic principles of popular dimensionality reduction algorithms and compare their strengths and weaknesses.
    • Demonstrate how computational analysis enhances the power of high parameter flow cytometry.

    Who Should Attend

    Cytometry practitioners and data analysts who encounter multidimensional datasets and employ algorithmic data analysis approaches or are interested in learning these methods

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 05/21/2021

    A CYTO U Webinar presented by Kevin Tsia, PhD Keywords: Imaging flow cytometry, high-throughput screening, single-cell analysis, biophysical cytometry

    About the Presenter

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    Kevin Tsia, PhD
    Professor
    University of Hong Kong

    Kevin Tsia received his PhD degree in electrical engineering at the University of California Los Angeles (UCLA) in 2009. He is currently a professor in the Department of Electrical and Electronic Engineering and the program director of the Biomedical Engineering Program at the University of Hong Kong (HKU). His research interest covers a broad range of subject matters including ultra-fast optical imaging for imaging flow cytometry and cell-based assay high-speed in-vivo brain imaging computational approaches for single-cell analysis. In 2012, he received the Early Career Award by the HK Research Grants Council in Hong Kong. He also received the Outstanding Young Research Award in 2015 at HKU as well as the 14th Chinese Science and Technology Award for Young Scientists in 2016. He is currently the RGC Research Fellow. He holds four-granted and four-pending US patents on ultrafast optical imaging technologies. He is co-founder of a start-up company commercializing the high-speed microscopy technology for cancer screening and treatment monitoring applications.

    Webinar Summary

    It has long been recognized that the association between the molecular genetic landscape that instructs expression of proteins and macromolecules in single cells is intrinsically linked with their biophysical properties (e.g., cell morphology, size, mass, force etc.). A growing body of evidence shows that the label-free assessment of biophysical properties of cells is an effective (or even more accurate) descriptor of cellular heterogeneity, compared to the conventional fluorescence markers, at single-cell precision. Furthermore, how molecular signatures translate into the emergent cellular biophysical properties has not been fully understood. Only with the recently advanced techniques can we now start to investigate this link.

    This webinar will introduce how the synergism among single-cell imaging, microfluidics, and deep learning allows us to overcome the current limitations of single-cell biophysical phenotyping (in both instrumentation integration and new data analytic strategies). Specifically, a few high-throughput, deep-learning-powered imaging techniques will be described, as well as cytometry pipelines developed in our laboratory over the past few years. These platforms allow researchers to significantly scale the single-cell biophysical phenotyping throughput (beyond millions of cells) and enrich the phenotyping content by integrating with biochemical cell-based assay in a single-platform. Pushing the limit of biophysical phenotyping specificity and sensitivity, these techniques have been successfully employed to a number of biological research and clinical applications, including rare cancer cell detection in mouse blood, cancer cell sub-typing, targeted-drug sensitivity prediction, and so on.

    Learning Objectives

    • Recent advances in biophysical cytometry, especially label-free single-cell imaging, that enables studies of cellular heterogeneity at the levels of throughput, precision, specificity, and sensitivity that were once inconceivable.
    • Advanced techniques, involving synergism among microfluidics, imaging and deep learning, that allow us to investigate deeper the link between molecular signatures and the emergent cellular biophysical properties. 

    Who Should Attend

    Biomedical scientists and engineers working on developments of cytometry platforms, single-cell imaging technologies, and single-cell analysis. 

    CMLE Credit: 1.0

  • Contains 6 Product(s)

    Cytometry panels packaging.

    • The Road to a High-Resolution 40-Color Flow Cytometry Immunophenotyping Panel
    • Designing Panels for the Study of Hematopoietic Stem Cells
    • Panel Design - A Practical Guide for Successful Fluorescent Cytometry Panels from 10-40 Parameters
    • Predicting the Best Resolution and Sensitivity in Panel Development and Reducing Inter-instrument Variability in Flow Cytometry
    • 16-Color Panel to Measure Inhibitory Receptor Signatures from Multiple Human Immune Cell Subsets
    • Step-by-Step Multi-parameter Panel Design 
  • Contains 3 Component(s), Includes Credits

    A CYTO U Webinar presented by Maria C. Jaimes, MD Keywords: High-dimensional flow cytometry, spectral flow cytometry, OMIP, Panel development

    About the Presenter

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    Maria C. Jaimes, PhD
    VP Technical Application Support
    Cytek Biosciences, Inc.

    Dr. Maria Jaimes earned her MD degree at the Universidad Javeriana in Colombia. Dr. Jaimes completed her postdoctoral training at Stanford University in the Department of Microbiology and Immunology. During her postdoc, she focused on characterizing the immune responses to both rotavirus and influenza viruses after natural infection and immunization. In 2005, Dr. Jaimes joined BD Biosciences, where she worked on different aspects of quality assurance and standardization of flow cytometry assays. In 2015, Dr. Jaimes joined Cytek Biosciences. She is part of the R&D team credited for developing the Aurora Full Spectrum Cytometer and has overseen the instrument characterization, verification, and development of multicolor applications. Besides her responsibilities within the R&D team, Dr. Jaimes leads the Technical Applications Support team worldwide.

    Webinar Summary

    This webinar will cover OMIP-069 (published in Cytometry Part A in August 2020), the first 40-color fluorescent panel using full spectrum flow cytometry to broadly phenotype much of the cellular composition of the human peripheral immune system. The panel in this OMIP has been thoroughly optimized to ensure high-quality data and well-resolved populations, enabling the description of most canonical subsets of T cells, B cells, NK cells, monocytes, and dendritic cells. Dr. Jaimes will present the journey of the technology, panel design, protocol development, data QC/QA, and data analysis which led to the successful achievement of this 40-color immune profiling panel. 

    Learning Objectives

    • Understand the concepts behind full spectrum profiling.
    • Learn the necessary steps and tools for good panel design.
    • Develop expertise on how to troubleshoot and optimize a multicolor panel.
    • Learn to recognize high quality vs. compromised data and the potential sources and mitigation of errors. 

    Who Should Attend

    SRL staff/directors, immunologists, CRO staff, and full spectrum flow cytometer users.

    CMLE Credit: 1.0

  • Contains 5 Product(s)

    An overview of fluorescence cytometry

    • Single Cell Analysis of Autofluorescence Lifetime Images 
    • Multiplexed Fluorescence Microscopy Reveals Heterogeneity among Stromal Cells in Mouse Bone Marrow Sections
    • Evaluating Spectral Cytometry for Immune Profiling in Viral Disease
    • The Essentials of Optics and Fluorescence Microscopy for Cell Biology Applications (2013 Advanced Data Analysis Pre-Congress Course)
  • Contains 3 Component(s), Includes Credits

    A CYTO U Webinar presented by Paula Niewold, PhD and Thomas Ashhurst, PhD Keywords: spectral cytometry, unmixing, compensation, conventional cytometry, autofluorescence, panel design, spreading error, platform comparison, data analysis

    About the Presenters

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    Paula Niewold, PhD
    Postdoctoral Researcher
    Department of Infectious Diseases
    Leiden University Medical Centre

    Dr. Paula Niewold is an immunologist currently working as a postdoctoral researcher at the Department of Infectious Diseases at the Leiden University Medical Centre. She is interested in host-pathogen interactions and how they impact the outcome of disease. She has studied these interactions in models of cerebral malaria, West Nile virus encephalitis, psoriasis, and tuberculosis using high-dimensional flow, mass, and imaging mass cytometry. She is an ISAC Marylou Ingram Scholar.

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    Thomas Ashhurst, PhD
    Immunologist and High-Dimensional Cytometry Specialist
    Sydney Cytometry Facility
    University of Sydney

    Dr. Thomas Ashhurst is an immunologist and high-dimensional cytometry specialist with the Sydney Cytometry Facility at the University of Sydney. He develops and applies a range of single-cell cytometry technologies and computational analysis tools to map dynamic immune responses over time, space, and disease. In particular, he applies these approaches to the study of immunology and infectious disease, including emerging pathogens such as COVID-19, Zika virus encephalitis, and West Nile virus encephalitis. He is an ISAC Marylou Ingram Scholar.

    Webinar Summary

    In conventional fluorescence cytometry, each fluorophore in a panel is measured in a target detector, through the use of wide band-pass optical filters. In contrast, spectral cytometry uses a large number of detectors with narrow band-pass filters to measure a fluorophore's signal across the spectrum, creating a more detailed fluorescent signature for each fluorophore. The spectral approach shows promise in adding flexibility to panel design and improving the measurement of fluorescent signal. However, few comparisons between conventional and spectral systems have been reported to date. Here we present our findings comparing conventional and spectral approaches to cytometry—including comparisons of compensation and unmixing—and evaluate the use of spectral cytometry for immune profiling in viral diseases.

    Learning Objectives

    • Gain an understanding of the essential differences between conventional and spectral approaches to cytometry.
    • Appreciate the differences between compensation and spectral unmixing.
    • Consider applications for spectral cytometry in the context of immunological studies.

    Who Should Attend

    Researchers and technical staff who utilize flow, spectral, or mass cytometry in their work.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 03/04/2021

    A CYTO U Webinar presented by Pablo Engel, PhD Keywords: antibodies, reagent validation, reproducibility

    About the Presenter

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    Pablo Engel, PhD
    Professor of Immunology
    University of Barcelona

    Pablo Engel obtained his MD and PhD from the University of Barcelona. After training as a postdoctoral fellow at the Dana-Farber Cancer Institute of Harvard Medical School, Pablo became an assistant professor in the Department of Immunology at Duke University School of Medicine. He is currently a professor of immunology and head of the Immunology Unit in the Department of Biomedical Sciences at the University of Barcelona. His research is focused on lymphocyte cell-surface molecules and their role in the regulation of immune responses. He is also an expert in the production and characterization of monoclonal antibodies. In addition to his research, Pablo is Secretary General of the European Federation of Immunological Societies (EFIS).

    Webinar Summary

    The presentation will highlight the current reproducibility crisis, specifically reflecting on the effect of poorly validated antibodies on the research. The main causes of antibody failure will also be reviewed, as well as a basic antibody validation protocol for flow cytometry. Several solutions will be discussed to solve the problem of reproducibility. 

    Learning Objectives

    • To understand the urgent need of antibody validation.
    • To gain knowledge about basic antibody validation protocols.

    Who Should Attend

    Scientists and technologist that use monoclonal antibodies of flow cytometry.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 12/10/2020

    A CYTO U Webinar presented by Dr. Peter Bajcsy, Dr. Nathan Hotaling, and Dr. Sreenivas Bhattiprolu Keywords: panel design, OMIP, history, autofluorescence, antibody titration

    About the Presenters

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    Dr. Peter Bajcsy
    Project Lead
    National Institute of Standards and Technology (NIST)
    Information Technology Laboratory (ITL)   

    Peter Bajcsy received his PhD in electrical and computer engineering in 1997 from the University of Illinois at Urbana-Champaign and an MS in electrical and computer engineering in 1994 from the University of Pennsylvania. He worked for machine vision, government contracting, and research and educational institutions before joining National Institute of Standards and Technology (NIST) in June 2011. At NIST, he is leading a project focusing on the application of computational science in metrology, specifically live cell and material characterization at very large scales.

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    Dr. Nathan Hotaling
    Vice President
    Data Science Solutions
    Information Technology Research Branch
    National Center for Advancing Translational Science-NIH

    Dr. Nathan Hotaling is a senior data scientist within the Information Resources Technology Branch at National Center for Advancing Translational Science-NIH (NCATS). He received his PhD in biomedical engineering from the Georgia Institute of Technology and an MS in clinical research from Emory University. After his PhD, Nathan conducted postdoctoral research in a joint project between the National Institute of Standards and Technology (NIST) and the National Eye Institute (NEI), where he began to develop a platform to analyze high-content image datasets collected for cell bio-manufacturing. This work led to his transition to his current position where he oversees the development of a scalable image analysis platform to non-invasively assess cell and tissue architecture, functionality, phenotype, consistency, and viability. Using this platform with novel machine learning and deep learning techniques, he intends to unlock the next “-omics” of cell analysis, "Vis-omics," for both research and clinical projects. He has co-authored 22 journal papers, two book chapters, and three patents.

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    Dr. Sreenivas Bhattiprolu
    Head of Digital Solutions
    Research Microscopy Solutions
    ZEISS

    Dr. Sreenivas Bhattiprolu (Sreeni) is the head of digital solutions at Carl Zeiss Microscopy. His team focuses on solving tough microscopy challenges by leveraging the latest advancements in digital technology and artificial intelligence. Sreeni has over 25 years of experience in microscopy in a variety of fields including life sciences, materials sciences, geosciences, electronics, and semiconductor technologies. Sreeni received his PhD in materials sciences and engineering from Michigan Technological University and earned his master’s degree in physics from the University of Hyderabad.

    Webinar Summary

    According to a recent survey by Nature, more than 70 percent of researchers have tried and failed to reproduce another scientist's experiments. Many factors contributing toward irreproducibility can be addressed via automation and collaborative work. Computer cloud provides the right infrastructure to automate image analysis tasks, especially for resource-intensive applications. Cloud's importance in regards to collaborative work has also been heightened in light of the COVID-19 pandemic due to its accessibility from any location and any networked device. This webinar further explains the benefits of cloud-based image analysis and introduces the audience to two platforms that facilitate automation and collaborative work with microscopy images. These two platforms have been independently developed by ZEISS and NIST/NIH, respectively. The presentation will go over the main features of the two platforms that run computational workflows formed by software containers that are interoperable. The discussion will include a variety of commercial and open source aspects in developing and using such platforms by the ISAC community.   

    Learning Objectives

    1. Why web/cloud-based image software solutions? How does your software work? 
    2. What are the advantages/disadvantages of your platform compared to traditional/existing approaches? When is best to think about using WIPP or APEER to solve quantitative imaging problems? 
    3. What does your software do? What functionality does it have? What is the best way for a potential user or group to get started using your platform (i.e., downloading, required hardware, necessary expertise, etc.)? 
    4. Considering that web/cloud-based platforms are a newer tool with a much smaller user population compared to packages such as ImageJ, Matlab, and Cell Profiler, what do you see as the future for these platforms? How will they grow? What are your plans for rolling out your platform and increasing adoption rates?


    CMLE Credit: 1.0

  • Contains 3 Product(s)

    Flow cytometry and hematology package.

    • Designing Panels for the Study of Hematopoietic Stem Cells
    • Flow Cytometric Analysis of Endothelial Colony Forming Cells and Hematopoietic Progenitor Cells in Lung Vascular Disease by Kewal Asosingh and Imaging Flow Cytometry in the Study of Immune Cell Functions by Andrew Filby
    • Flow Cytometry Analysis of Human Hematopoietic Progenitors in Cardiovascular Disease
  • Contains 4 Component(s), Includes Credits Recorded On: 10/29/2020

    A CYTO U Webinar presented by Dr. Steffen Schmitt and Dr. Marcus Eich Keywords: panel design, OMIP, history, autofluorescence, antibody titration

    About the Presenters

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    Dr. Steffen Schmitt
    German Cancer Research Center (DKFZ)  

    Steffen Schmitt studied biology at the Ruprecht-Karls-University Heidelberg and finished his PhD with a concentration in immunology. Subsequent to his postdoc, he developed the flow cytometric service at the Center for Natural and Medical Sciences (NMFZ) at Johannes-Gutenberg-University in Mainz. Since 2007 Steffen has headed the Flow Cytometry Core Facility of the German Cancer Research Center (DKFZ) in Heidelberg.

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     Dr. Marcus Eich
     Hi-Stem gGmbH

    Marcus Eich studied biology at the Technical University Darmstadt and earned his PhD in 2011 in toxicology at the University Medical Center Mainz. After an additional postdoc in Mainz in 2015, he joined the HI-STEM team and the Flow Cytometry Core Facility of the German Cancer Research Center (DKFZ) in Heidelberg.

    Webinar Summary

    The hematopoietic system is a very fascinating system to study as it can re-populate an entire system starting from just one cell. This webinar gives an overview of the targeted subpopulations of the hematopoietic system and how different experimental tasks and setups in one panel were combined. At the end, issues that occurred during the panel design are discussed and a short outlook is presented about how to further improve the panel.

    Learning Objectives

    • Understand the hematopoietic system from a backbone panel to specialized subpopulations, combining different experimental setups in panel design.
    • Apply tips and tricks for this panel.

    Who Should Attend

    PhD students, postdocs, and technicians involved in hematopoietic stem cell research.

    CMLE Credit: 1.0

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  • Contains 4 Component(s), Includes Credits Recorded On: 10/22/2020

    A CYTO U Webinar presented by Thomas Liechti, PhD Keywords: panel design, OMIP, history, autofluorescence, antibody titration

    About the Presenter

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    Thomas Liechti
    Postdoctoral Researcher
    National Institutes of Health (NIH)

    Thomas Liechti obtained his PhD in immunology and microbiology at the University of Zurich in 2017 and is currently a postdoctoral researcher in Mario Roederer’s group at the Vaccine Research Center of the National Institutes of Health. His main interest is high-dimensional flow cytometry and human immunology. During his postdoctoral training, he established a 28-color flow cytometry sample processing and analysis pipeline to assess the contribution of genetic and environmental factors to human immune homeostasis. 

    Webinar Summary

    High-dimensional flow cytometry enables the characterization of the human immune system at unprecedent depth and at a population-wide scale. However, flow cytometry panel design poses many caveats and requires careful testing of several reagents and panel iterations to find optimal combinations of reagents with the best possible performance (i.e., low spill-over spreading and high resolution). 

    This webinar is the second part of a back-to-back webinar with Florian Mair. In the first part, Florian will cover the development and common caveats of high-parameter flow cytometry panels. In the second part of this webinar series, Thomas will be talking about more specific caveats he encountered during the development of panels described in OMIP-051, -058 and -060.

    Large immunophenotyping studies including thousands of samples and require several experiments over an extended period of time, which can be prone to inter-assay variation. Thomas will discuss strategies for building an optimal staining and sample processing pipeline for an immunophenotyping study. This process includes over 3,000 samples as well as tracking and reducing technical (instrument performance) and experimental variation. In addition, Thomas will give some outlook into data pre-processing including automated exclusion of data variation and compensation.

    Learning Objectives

    • An understanding for tackling high-dimensional immunophenotyping assays on a variety of platforms, thus minimizing trial-and-error experiences and wasted experiments
    • Review specific strategies for generating reproducible and high-quality data in large patient cohorts.

    Who Should Attend

    Anyone with an interest in efficient panel design: immunologists, scientists of any field doing polychromatic flow cytometry, SRL users, and SRL leaders.


    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 10/21/2020

    A CYTO U Webinar presented by Florian Mair, PhD Keywords: panel design, antibody titration, compensation, thawing, PBMCs

    About the Presenter

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    Florian Mair
    Cytometry Specialist
    Fred Hutchinson Cancer Research Center

    Florian graduated with a PhD from the University of Zurich, Switzerland in 2014 and is currently working at the Fred Hutchinson Cancer Research Center as an immunologist. During the past decade, he has been involved extensively with different cytometry platforms (conventional, spectral, and mass cytometry) as well as scRNA-seq technique. Florian is interested in applying novel analysis approaches for single cell data. He has been actively engaged in teaching flow cytometry courses including systematic panel design and analysis of high-dimensional cytometry experiments. Florian is currently an ISAC Marylou Scholar.

    Webinar Summary

    Over the past decade, technical improvements and new reagents have permitted fluorescent-based flow cytometry assays to measure up to 40 parameters. These complex assays require robust controls and thorough experimental planning, but there are currently few resources that provide a systematic approach for reliable panel design. Also, historical notions as to how fluorophores and controls should be chosen are sometimes at odds with the reality of modern panel design.

    In this webinar, we will provide a practical guide for successful fluorescent panel design for any complex panel from 10–⁠40 (or more) parameters, both for conventional compensation-based as well as spectral cytometry.

    Specifically, we will cover the following topics:

    • Brief overview of signal detection in conventional and spectral flow cytometers.
    • The concept and underlying cause of spreading error (SE).
    • How the spillover spreading matrix (SSM) can be efficiently used to guide panel design.
    • Relevant relationships between SE, fluorophore brightness, and antigen expression level.
    • Step-by-step approaches toward building a new panel.
    • An overview of essential controls and typical caveats.

    This webinar is back-to-back with a webinar by Thomas Liechti, who will be talking about how to best design and use complex panels for large study cohorts (100s-1000s of samples) including the use of appropriate controls and analysis approaches.

    Learning Objectives

    • An understanding for tackling high-dimensional immunophenotyping assays on a variety of platforms, thus minimizing trial-and-error experiences and wasted experiments
    • Review specific strategies for generating reproducible and high-quality data in large patient cohorts.

    Who Should Attend

    Anyone with an interest in efficient panel design: immunologists, scientists of any field doing polychromatic flow cytometry, SRL users, and SRL leaders.


    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 10/13/2020

    A CYTO U Webinar presented by Sara De Biasi, PhD Keywords: metabolites, immunology, mitochondria, T cells

    About the Presenter

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    Sara De Biasi
    Postdoctoral Research
    University of Modena and Reggio Emilia

    Sara De Biasi obtained her PhD in clinical and experimental medicine (immunology) from the University of Modena and Reggio Emilia in 2013. Most of her work focuses on the variability of adaptive immune response in HIV, autoimmune disease, and cancer. For the last two years, she has studied T cell metabolism with particular interest in mitochondria. Dr. De Biasi is an ISAC Marylou Ingram Scholar and a member of the CYTO U Task Force. 

    Webinar Summary

    This webinar will focus on methods to investigate metabolic changes and mithochondria functionality of T cell subpopulations. Data obtained studying T cells in a group of progressive multiple sclerosis patients will be discussed.

    Learning Objectives

    Upon completion of this webinar, you will be able to:

    • Evaluate mitochondria phenotype and functionality.
    • Evaluate T cell metabolism. 
    • Evaluate differences among functionally for different T cells.

    Who Should Attend

    Researchers who want to learn how to dissect metabolic changes, mitochondrial phenotype, and functionality in human T cells in different pathophysiological conditions.

    CMLE Credit: 1.0

  • Contains 3 Component(s) Recorded On: 06/23/2020

    ISAC General Business Meeting 2020

    Introduction

    Hear from ISAC's President Andrea Cossarizza, President-Elect Jonni Moore, Treasurer Jessica Houston, and Executive Director Michelle Butler on the state of the society, its finances, and future during ISAC's annual business meeting on June 23.

    Agenda

    1. Call to Order - Andrea Cossarizza
    2. Approval of June 26, 2019 - Andrea Cossarizza CYTO 2019 General Business Meeting Minutes
    3. Treasurer's Report - Jessica Houston
    4. ISAC's Leadership Development Program - Andrea Cossarizza
    5. ISAC's State of the Society - Andrea Cossarizza
    6. Introduction of New ISAC Officers and Councilors - Andrea Cossarizza
    7. ISAC – Moving to the Future - Jonni Moore
    8. Old Business - Jonni Moore
    9. New Business - Jonni Moore
    10. Moderated Questions & Answers from Audience - Michelle Butler
    11. Adjournment - Jonni Moore

    Click here to view the minutes of the 2019 ISAC General Business Meeting.

  • Contains 3 Component(s), Includes Credits Recorded On: 05/28/2020

    A CYTO U Webinar presented by Andrea Cossarizza, PhD Keywords: immunophenotyping, viral infection, SARS-CoV-2, high-dimensional cytometry

    About the Presenter

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    Andrea Cossarizza
    Full Professor of Pathology and Immunology
    University of Modena and Reggio Emilia

    Andrea Cossarizza is a full professor of pathology and immunology, vice president of the Faculty of Medicine at the University of Modena and Reggio Emilia, and former president of the International Society for Advancement of Cytometry (ISAC). He has been studying the molecular and cellular basis of immune system-based diseases for 35 years. He is currently at the forefront of the fight against COVID-19 and has provided the first contributions regarding the importance of cytometry in understanding the immune response to SARS-CoV-2. As of May 2020, he has published 332 papers in peer-reviewed journals, has an H index of 79, and received over 33,000 citations.

    Webinar Summary

    The immune system is heavily involved in the pathogenesis of COVID-19 and its activation (which includes the so-called cytokine storm) and is the main force that drives the course of the infection. This webinar will present the most recent data on main changes that occur among different lymphocyte populations, along with functional analysis of T cells, and discuss current and possible therapeutic approaches.

    Learning Objectives

    • Describe the main changes that occur in the immune system during different phases of the infection with SARS-CoV-2, the virus that causes COVID-19.
    • Identify cytometric techniques that have been used to better understand changes that occur in the immune system during the infection and its recovery.

    Who Should Attend

    Researchers, clinicians, laboratory managers, and personnel involved in the fight against SARS-CoV-2 who want to know the latest discoveries on the role of the immune system during COVID-19.

    CMLE Credit: 1.0

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  • Contains 4 Component(s), Includes Credits Recorded On: 01/08/2020

    A CYTO U Webinar presented by Keisuke Goda, PhD Keywords: Cell sorting, imaging, advanced technology

    About the Presenter

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    Keisuke Goda, PhD
    Professor
    Department of Chemistry at the University of Tokyo

    Dr. Keisuke Goda is a professor in the Department of Chemistry at the University of Tokyo, an adjunct professor in the Institute of Technological Sciences at Wuhan University, and an adjunct professor in the Department of Bioengineering at UCLA. He obtained a BA degree from UC Berkeley summa cum laude in 2001 and a PhD from MIT in 2007, both in physics. At MIT, he worked on the development of gravitational-wave detectors in the LIGO group which led to the 2017 Nobel Prize in Physics. After several years of work on high-speed imaging and microfluidics at UCLA, he joined the University of Tokyo as a professor. His research group focuses on the development of serendipity-enabling technologies based on molecular imaging and spectroscopy together with microfluidics and computational analytics to push the frontier of science. He is an associate editor of Cytometry Part A and APL Photonics. He has published over 350 journal and conference papers, filed over 30 patents, and received numerous awards such as Japan Academy Medal, JSPS Prize, and Analytical Chemistry Young Innovator Award.

    Webinar Summary

    The advent of intelligent Image-Activated Cell Sorting (iIACS) has enabled high-throughput intelligent image-based sorting of single live cells or cell clusters with unique morphochemical features that are difficult to discern when compressing these spatial data into intensity signals in fluorescence-activated cell sorting (FACS) [Nitta et al., Cell 175, 266-276 (2018)]. iIACS is an on-chip microfluidic technology that builds on a seamless integration of a high-throughput fluorescence microscope, cell focuser, cell sorter, and deep neural network on a hybrid software-hardware data management architecture, thereby providing the combined merits of optical microscopy, FACS, and deep learning [Isozaki et al., Nature Protocols 14, 2370-2425 (2019)]. Therefore, iIACS serves as an essential part of holistic single-cell analysis by providing direct connections between population-level analysis (flow cytometry), cell-level analysis (microscopy), and gene-level analysis (sequencing) of sorted cells. In this webinar, Keisuke will give a tutorial about the principles and applications of iIACS and compare the usability of iIACS with other technologies such as fluorescence-activated cell sorting (FACS), imaging flow cytometry (without sorting), and image-based cell pickers.

    Learning Objectives

    • Explore the principles and applications of the iIACS technology.

    Who Should Attend

    FACS developers/users, imaging flow cytometry developers/users, single-cell analysis researchers, etc.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 12/17/2019

    A CYTO U Webinar presented by Geoffrey Lyon, MPH Keywords: aerosol evaluation, risk assessment, Glo-Germ, Dragon green bead, cyclex-D

    About the Presenter

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    Geoffrey Lyon, MPH
    Yale Flow Cytometry Facility Manager/BSL-3 Sort Operator
    Yale University

    Geoffrey Lyon is a Flow Cytometry Facility manager/BSL-3 Sort operator at Yale University. One of his most prominent career achievements is discovering cell sorter AMS filter failures. He is a member of the ISAC Biosafety Committee, a former ABSA International Course presenter, a Yale BSL-3 Subcommittee member, and a BSL-3 Certified Researcher and Trainer.

    Webinar Summary

    This webinar explores the various biosafety aspects of high-speed cell sorting. The emphasis of this presentation is the evolution and development of aerosol containment testing including the new ISAC standard that uses a novel impactor and microspheres to test containment. This new assay can be utilized to test sorters housed inside or outside of a biosafety cabinet.

    Learning Objectives

    • Examine the hazards and risks associated with cell sorting along with a detailed explanation of the latest containment testing method.
    • Describe how to implement the new testing method into existing safety protocols to ensure cells sorters have adequate aerosol containment.    

    Who Should Attend

    Anyone who uses high-speed cell sorters, sort operators, health and safety professionals, or anyone interested in biosafety.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 11/19/2019

    A CYTO U Webinar presented by Alex Walsh, PhD Keywords: Optical imaging, 2-photon imaging, cancer therapy, Autofluorescence

    About the Presenter

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    Alex Walsh, PhD
    Assistant Professor in Biomedical Engineering
    Texas A&M University

    Dr. Walsh completed her Ph.D. at Vanderbilt University, where she developed an autofluorescence lifetime-based assay for determining the optimal cancer treatment strategy for individual patients. As a postdoc at the Air Force Research Lab, Dr. Walsh used optical techniques to investigate infrared-light activation and inhibition of action potential propagation in neurons. Currently, Dr. Walsh is an assistant professor in the Biomedical Engineering Department at Texas A&M University.

    Webinar Summary

    Fluorescence lifetime imaging (FLIM) of the endogenous fluorophores, NAD(P)H and FAD (co-enzymes of metabolic reactions), provides a label-free method to quantify cellular metabolism. This webinar will review multi-photon fluorescence lifetime imaging methods, single cell segmentation, and intra-population heterogeneity analysis. Examples will be shown for drug response in breast cancer organoids and activation of T cells.

    Learning Objectives

    • Define fluorescence lifetime and time-correlated single photon counting imaging methods.
      Interpret label-free FLIM images of NAD(P)H and FAD.
    • Discuss segmentation and single-cell analysis techniques.

    Who Should Attend

    Anyone interested in label-free imaging, fluorescence lifetime imaging, or single-cell analysis.

    CMLE Credit: 1.0

  • Contains 3 Product(s)

    Data analysis series package.

    • Data Analysis Rigor and Reproducibility, Part 1: Experimental Design
    • Data Analysis Rigor and Reproducibility, Part 2: Analysis Tools
    • Data Analysis Rigor and Reproducibility, Part 3: Publishing Flow Cytometry Data
  • Contains 3 Component(s), Includes Credits Recorded On: 10/22/2019

    A CYTO U Webinar presented by Aja Rieger, PhD & Andrew Filby, PhD Keywords: publication, visualisation, guidelines, MIFlowCyt, Flow Cytometry, Mass Cytometry, Genomic Cytometry

    About the Presenters

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    Aja Rieger, PhD
    Flow Core Manager
    University of Alberta

    Dr. Rieger graduated from the University of Alberta with a BSc honors in immunology and infection. She then obtained an MSc in neuroimmunology from McGill University. Following this, Aja returned to the University of Alberta for her PhD studies in comparative immunology, researching the role of macrophages in initiating and resolving inflammation in goldfish. She then moved to University of California-Berkeley for her postdoctoral fellowship in neuroimmunology. In her current role as the flow core manager at the University of Alberta, Faculty of Medicine and Dentistry, Aja oversees the operations of both the Flow Cytometry Facility and the High Content Analysis Core. She manages a team of cytometry technologists, with a specialty in imaging flow cytometry assay development. Aja is currently an ISAC SRL Emerging Leader.

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    Andrew Filby
    Director of the Newcastle University Cytometry and Single Cell Core Technology Unit
    Newcastle University

    Dr. Filby graduated summa cum laude from the University of Huddersfield with a first class honors in biochemistry. After graduating, he undertook a Ph.D. at the National Institute for Medical Research (NIMR) in Mill Hill, London. He worked on the Src family kinases LCK and Fyn in adaptive immunity, obtaining his PhD in molecular and cellular immunology from University College London (UCL).  Dr. Filby remained in the immunological field at the NIMR, working as a postdoctoral researcher on models of retroviral infection. He then worked in the commercial sector before taking up the deputy head role of the cytometry core at the London Research Institute (now the Francis Crick). Dr. Filby is currently director of the Newcastle University Cytometry and Single Cell Core Technology Unit. He leads a dedicated team of cytometry specialists with the sole aim of developing and implementing comprehensive, cutting-edge cytometry methods for the wider research community at Newcastle University and beyond. A significant part of his focus is the development of novel cytometry-based techniques that have underpinned several high-profile publications in journals including Science (2012, 2017, and 2018), Cell (2013), and Nature (2018). His current research is focused on whether label-free imaging cytometry techniques can be used to refine or replace the need for directed probes in order to prove cellular identity.

     Webinar Summary

    This webinar will give an overview of the current guidelines for publishing flow data with a high level of rigor. We will discuss publication of both standard flow cytometry data, as well as imaging cytometry, mass cytometry, and genomic cytometry data sets.

    Learning Objectives

    • Understand MIFlowCyt guidelines for publishing flow cytometry data.
    • Use best practices for communicating cytometry data in publications.
    • Review key points to include in any methods section toward reproducibility.
    • Explore data repositories.

    Who Should Attend

    Anyone interested in publishing high-quality, rigorous flow cytometry data.

    CMLE Credit: .75

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  • Contains 3 Component(s), Includes Credits Recorded On: 10/16/2019

    A CYTO U Webinar presented by Nicole Poulton, PhD & Rachael Sheridan Keywords: Aquatic samples, Biomedical samples, small particals, Nuclear,isolation, Instrument setup, Dilution

    About the Presenters

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    Nicole Poulton, PhD
    Director of the Facility for Aquatic Cytometry
    Bigelow Laboratory for Ocean Sciences

    Nicole Poulton is a research scientist and director of the Facility for Aquatic Cytometry at Bigelow Laboratory for Ocean Sciences, in East Boothbay, Maine. Her research uses both flow and imaging cytometry to identify and examine viruses, bacteria, and plankton from natural environments. She works primarily with samples from natural communities, ranging from lakes and oceans to hyper-saline ponds, sediments, soil, and mineral rich hot springs. She is an active educator and trains cytometrists, students, and scientists interested in learning aquatic and environmental cytometric techniques. Nicole received her PhD from the Massachusetts Institute of Technology/Woods Hole Oceanographic Institution Joint Program.

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    Rachael Sheridan
    Director of the Flow Cytometry Core Facility
    Van Andle Research Institute

    Rachael Sheridan is the director of the Flow Cytometry Core facility at the Van Andle Research Institute in Grand Rapids, Michigan. Her core supports a wide array of biomedical research ranging from immunology and metabolism to neurodegenerative disease and cancer. She works with samples originating from multiple tissue types as either whole cells or isolated nuclei, and she is always excited to try something new. Before moving to Grand Rapids, Rachael trained at the University of Wisconsin—Madison Carbone Cancer Center Flow Cytometry Core where she discovered her passion for flow cytometry and education.  

    Webinar Summary

    Life in a Shared Resource Flow Cytometry Laboratory (SRL) is always dynamic. In addition to routine samples, we are often faced with challenging and unique samples. In research settings these could be anything from subcellular organelles, such as nuclei and mitochondria, debris-ridden tissue preps, or non-mammalian organisms including plant cells and plankton, as well as bacteria and viruses. Each of these samples present unique challenges to the SRL cytometrist. In this tutorial, the presenters will discuss and present experiences working with these samples in both a biomedical and aquatic cytometry core facility and provide some approaches and tips to keep in mind when you confront these types of samples.  

    This webinar will address the following:

    • What types of samples can be analyzed by flow cytometry (biomedical to environmental)?
    • Why are SRLs observing more challenging samples?
    • How do operators prepare samples for cytometric analysis?
    • What steps should be considered during instrument setup?
    • Is auto-fluorescence a friend or foe?

    Learning Objectives

    This webinar and discussion will provide participants with a better understanding of how to handle and prepare for different types of samples as the biomedical field expands, as well as use of core facility changes. We will provide a link to a "tips and tricks" webpage addressing how to handle a variety of samples. 

    Who Should Attend

    SRL Core employees and researchers interested in working with non-traditional samples within a research or core facility setting.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 09/24/2019

    A CYTO U Webinar presented by Sofie Van Gassen, PhD Keywords: visualisation, dimensionality reduction, automated gating, clustering, FlowSOM, quality control

    About the Presenter

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    Sofie Van Gassen, PhD
    Postdoctoral Researcher
    Gent University Center for Inflammation Research

    Sofie Van Gassen received her MS in computer science from Ghent University in 2013 and her PhD in computer science engineering from Ghent University in 2017. During her PhD, she developed machine learning techniques for flow and mass cytometry data. She is an ISAC Marylou Ingram Scholar. As a postdoc she is further developing and improving machine learning techniques for single cell data in the DaMBi group (VIB - UGent Center for Inflammation Research).

    Webinar Summary

    Some of the current data analysis tools will be presented including tools for visualization (e.g., SPADE, tSNE, UMAP), automated gating (e.g., flowDensity, flowLearn), and population discovery (e.g., Citrus, FlowSOM, CellCNN). Detailed pros and cons of these methods will be highlighted along with a discussion on how to pick a good tool.

    Learning Objectives

    •     Learn about the dimensionality reduction algorithms, clustering algorithms, and population discovery tools.
    •     Discuss guidelines and learn how to select which tool is best depending on a given situation.

    Who Should Attend

    Anyone exploring analysis tools they could apply to their cytometry data.

    CMLE Credit: .75

  • Contains 3 Component(s), Includes Credits Recorded On: 09/17/2019

    A CYTO U Webinar presented by Alex Skovsbo Jørgensen Keywords: Cell death, Annexin V, Necrosis, apoptosis, fixable viability dye, TUNEL assay, DNA damage, membrane integrity, DNA intercalating dye, membrane potential

    About the Presenter

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    Alex Skovsbo Jørgensen
    Assistant Professor
    Department of Health Science and Technology
    Aalborg University

    Alex is an assistant professor in the Department of Health Science and Technology at Aalborg University. His research focus is using machine learning and image analysis within the domain of digital pathology. His current research topics of interest within digital pathology is automated cancer detection and grading, artificial intelligence, and quality assessment of staining protocols. 

    Webinar Summary

    Breast cancer is the most frequent cancer among women worldwide. Ki67 can be used as an immunohistochemical pseudo marker for cell proliferation to determine how aggressive the cancer is and thereby the treatment of the patient. No standard Ki67 staining protocol exists, resulting in interlaboratory stain variability. Therefore, it is important to determine the quality control of a staining protocol to ensure correct diagnosis and treatment of patients. Currently, quality control is performed by the organization NordiQC that use an expert panel-based qualitative assessment system. However, no objective method exists to determine the quality of a staining protocol.

    Learning Objectives

    •     Understand the challenges of staining quality assessment.
    •     Use cell lines for assessment of stain quality.
    •     How to use image analysis and machine learning for quality assessment of staining protocols.
    •     Understand validation challenges.

    Who Should Attend

    Pathologists and engineers within medial image analysis and machine learning.

    CMLE Credit: 1.0

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  • Contains 3 Component(s), Includes Credits Recorded On: 08/15/2019

    A CYTO U Webinar presented by Dagna Sheerar, SCYM Keywords: Reproducability, Statistical power, FMO, MIFlowCyt, Stain index

    About the Presenter

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    Dagna Sheerar, SCYM
    Manager
    University of Wisconsin Carbone Cancer Center (UWCCC) Flow Lab

    Dagna Sheerar has been working in flow cytometry shared resource laboratories since 2000, starting out as an assistant in organizing a BSL-3 cell sorting facility at the Immunology Services Laboratory of the Wisconsin National Primate Research Center. From there, she went on to manage the Flow Lab at the University of Western Ontario for three years. In 2006, Ms. Sheerar returned to the University of Wisconsin—Madison to work in the Carbone Comprehensive Cancer Center’s Flow Cytometry Laboratory. In 2011, she was hired as the manager of the UWCCC Flow Lab. Always an active member in the flow cytometry community, Ms. Sheerar is a member of the Steering Committee for the Great Lakes International Imaging and Flow Cytometry Association and a member of the ISAC Shared Resources Laboratory Educational Task Force. As manager of the UWCCC Flow Lab, Ms. Sheerar focuses on providing researchers with the tools and support to perform rigorous and reproducible flow cytometry assays in basic research and clinical research trials.

    Webinar Summary

    This webinar will outline the steps and considerations in designing a successful flow cytometry assay in the context of basic research and clinical research trials. We will focus on how best to minimize variables for a robust and reproducible assay, paying attention to producing data sets well suited to downstream computational data analysis platforms.

    Learning Objectives

    • Discuss the importance of working with biostatisticians in the early experimental planning stages.
    • Describe how to create criteria for sample inclusion/exclusion and building in room for sample loss.
    • Learn the importance of validating reagents and proper quality control and characterization of instrumentation.
    • Discuss how to create rigorous protocols and the importance of record keeping and annotation.
    • Expectations for the design, optimization, and standardization of assays and data analysis pipelines.

    Who Should Attend

    • Researchers using flow cytometry assays in the course of their research.
    • SRL staff supporting researchers in the design, optimization, standardization, and data analysis of these research projects.
    • Computational biologists and biostatisticians performing data analyses for large-scale flow cytometry based experiments.

    CMLE Credit: 1.0

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  • Contains 3 Component(s), Includes Credits Recorded On: 07/31/2019

    A CYTO U Webinar presented by Matthew Linden, PhD Keywords: platelet biology, cardiovascular disease, anticoagulant, sample collection

    About the Presenter

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    Matthew Linden, PhD
    Associate Professor of Haematology
    University of Western Australia

    Matthew is the Associate Professor of Haematology at the University of Western Australia, where he leads the development and delivery of haematology and cytometry education to train hospital scientists. Matthew’s research is in platelet biology and function. Working at the interface of the shared resource laboratory, discovery and translational research, Matthew has developed novel cytometry techniques for the measurement of blood platelets, and he has employed these in the development of new antiplatelet therapies. He is committed to advancing cytometry through strong, sustainable shared resource laboratories and cytometry education. Matthew is an ISAC Marylou Ingram Scholar and the president of the Australasian Cytometry Society.

    Webinar Summary

    Flow cytometry is a powerful and versatile tool which can be used to provide substantial phenotypic data on platelets including surface expression of functional receptors, bound ligands, expression of granule components, signal transduction, platelet-platelet aggregation, or interaction of platelets with leukocytes. Quantitative assessment of these parameters may facilitate the diagnosis of inherited or acquired platelet disorders, assist in the diagnosis of diseases associated with platelet activation, or help with the monitoring of safety and efficacy of anti-platelet therapy.

    Learning Objectives

    • Discuss the utility of platelet flow cytometry.
    • Describe the process by which platelet phenotype and function are measured.
    • Discuss the pre-analytical and analytical variables that can affect platelet data.

    Who Should Attend

    Clinical and research scientists who wish to measure platelet phenotype and function.

    CMLE Credit: 1.0

  • Contains 4 Product(s)

    Package for cell sorting in cytometry.

    • Cell Sorting: Fundamentals and Selected Novel Applications
    • Cell Sorting for Function and Viability
    • 3D Cytometry: Intelligent Image—Activated Cell Sorting
    • Current Standards in Flow Cytometry Cell Sorter Biosafety
    • Intelligent Image-Activated Cell Sorting: A Tutorial 
    • Optimizing SRL Performance: Boost your Cell Sorting Capacity
  • Contains 3 Component(s), Includes Credits Recorded On: 05/01/2019

    A CYTO U Webinar presented by Paul Tillberg, PhD Keywords: Advanced imaging, super resolution, diffraction limit, expansion

    About the Presenter

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    Paul Tillberg, PhD
    Fellow/Lab Head
    HHMI Janelia Research Campus

    Paul Tillberg completed a BS in electrical and materials engineering at UC Berkeley before going on to complete his PhD in electrical engineering at MIT, where he worked on technology development for the biological sciences. In the Boyden group, he conceived, designed, and co-led the development of Expansion Microscopy. He is currently a Fellow/Lab Head at the HHMI Janelia Research Campus, where he continues to develop and disseminate the expansion method in addition to other avenues of technology development for biology.

    Webinar Summary

    The expansion microscopy method improves the effective resolution of any optical microscope by ~4-fold by uniformly expanding biological specimens. Expansion is achieved by embedding the tissue in an ultra-swellable gel, followed by a few simple processing steps. The method is easy to adopt and well suited as a histology core facility offering, as it is compatible with existing antibody and fluorescent protein labelling protocols without modification.

    Learning Objectives

    • Discuss how to do expansion microscopy.
    • Describe the variants associated with expansion microscopy.
    • Learn the strengths and drawbacks to evaluate expansion microscopy for user specific applications.

    Who Should Attend

    Any biologist interested in probing the nanoscale structure of fixed tissue. Basic histology and imaging skills are useful for the smooth adoption of expansion microscopy.

    CMLE Credit: .75

  • Contains 3 Component(s), Includes Credits Recorded On: 04/15/2019

    A CYTO U Webinar presented by Anja E. Hauser, PhD Keywords: multiplex histology, quantitative, bone marrow

    About the Presenter

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    Anja Hauser, PhD
    Professor of Immune Dynamics and Intravital Microscopy
    Charité—Universitätsmedizin and Deutsches Rheumaforschungszentrum

    During her studies of veterinary medicine, Anja Hauser developed an interest in immunopathology and microscopy. As a graduate student, she identified factors which attract plasma blasts into the bone marrow and keep them alive in specialized niches within this tissue. During her postdoctoral work, she worked on the migration of germinal center B cells using intravital 2-photon microscopy. She is a professor of immune dynamics and intravital microscopy at the Charité—Universitätsmedizin and Deutsches Rheumaforschungszentrum in Berlin. In an interdisciplinary approach, her lab develops novel microscopy technologies to obtain a deeper insight in how the immune system functions. 

    Webinar Summary

    Anja will introduce the principles of multi-epitope ligand cartography (MELC), a method for multiplexed immunofluorescence microscopy, and explain its application for analyzing complex tissues and rare cell subsets. She will also give an overview on methods suitable to quantitatively analyze those complex multi-parametric image data.

    Learning Objectives

    • Describe MELC as a method for multiplexed immunofluorescence histology.
    • Learn what to consider when preparing tissues for MELC analysis, such as choosing marker panels and planning the sequential staining in the tissue
    • Learn about options for image analysis in order to extract quantitative information from multiplexed immunofluorescence histology.

    Who Should Attend

    Everyone who is interested in quantitative multiplexed image analysis and its applications in immunology.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 04/09/2019

    A CYTO U Webinar presented by Andre Olsson, PhD Keywords: fluorescent proteins, spectral overlap, spillover spreading, cellprofiler

    About the Presenter

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    Andre Olsson, PhD
    Research Associate
    Lee Grimes Lab

    Andre received his PhD in experimental hematology at the Faculty of Medicine, Lund University in Sweden. In graduate school he studied the function of the ETO (Eight Twenty One) homologues and found that they are involved in regulating hematopoietic progenitor maintenance and lineage differentiation. As a research associate at Lee Grimes lab, he has focused on understanding myeloid lineage decisions during homeostasis. Hematopoiesis is a great system for studying cells and the processes that instruct the cells how to function. Flow cytometry enables people to study cells at a single cell level, and thus is a very powerful tool to map and characterize cells going through differentiation.

    Webinar Summary

    Fluorescent protein labeling of specific genes combined with surface marker profiling can more specifically identify a cell population. The advent of facile genome engineering technologies has made the generation of gene-expression or fusion-protein reporters more tractable. While there are a number of fluorescent proteins available, their choice as reporter constructs is made difficult by the lack of data on how sensitivity and other factors are affected when two or more fluorescent proteins are combined. We characterize the detection sensitivity, spectral overlap, and spillover spreading of 13 monomeric fluorescent proteins to determine their utility in multicolor panels.

    Learning Objectives

    • Describe how to consider fluorescent protein detection sensitivity for fusion-protein studies.
    • Learn how spectral overlap and spillover spreading impacts which fluorescent proteins to combine in an experiment.
    • Discuss how experimental validation is key to successful panel design.

    Who Should Attend

    Anyone interested in using fluorescent proteins in their in vivo or in vitro research.

    CMLE Credit: .5

  • Contains 3 Component(s), Includes Credits Recorded On: 03/28/2019

    A CYTO U Webinar presented by Karel Drbal, PhD Keywords: dimensionality reduction, tSNE, UMAP, EmbedSOM, visualization

    About the Presenter

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    Karel Drbal, PhD

    Karel Drbal obtained a MSc in cell biology and a PhD in immunology from Charles University in 1991 and 2000, respectively. For most of his career he has been specialized in monoclonal antibody generation and characterization (MEM-series) by flow cytometry. After postdoctoral fellowships with Dr. Hannes Stockinger at Medical University Vienna and Dr. Vaclav Horejsi at Institute of Molecular Genetics of CAS, he held the CSO position in biotech enterprise Exbio Praha from 2010 to 2013. He is a teacher of immunology, systems biology, and cytometry, and in 2015 he launched the Lab of Molecular Dynamics of the Immune Response at Charles University.

    Webinar Summary

    Karel will introduced the state-of-the-art dimensionality reduction algorithms (SNE variants and UMAP) for unsupervised cytometry data analysis. Next, these algorithms will be compared side by side to the new EmbedSOM algorithm based on FlowSOM clustering approach. The importance of input data quality, the logic of clustering and embedding workflow and the output annotation will be described with the examples in R environment. Finally, the results of benchmarking datasets as well as few use cases analyzed by hierarchical dissection of the embedded data will be presented. To close, the comparison of manual workflow and the subjective visual perception of the output quality will be discussed, as well as the future directions of unsupervised versus supervised analysis beyond the field of cytometry will be outlined.

    Learning Objectives

    • Discuss the principle of the new EmbedSOM algorithm and the associated workflow.
    • Learn the differences between the embedded algorithms for data visualization.
    • Describe hierarchical dissection of complex datasets including statistical output.
    • Discuss the strengths and weaknesses of the unsupervised process.

    Who Should Attend

    • Clinical and research scientists collecting multidimensional data not only in the field of cytometry but also in the fields of microscopy, transcriptomics, and proteomics.
    • Computational scientists and bioinformatics core personnel interested in unsupervised data visualization.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 03/19/2019

    A CYTO U Webinar presented by Thomas N. Denny, M Phil, MSc Keywords: Regulatory, validation, FDA, legislation, laboratory developed test, in vitro clinical test

    PhAbout the Faculty

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    Thomas Dennny, M Phil, MSc
    Professor of Medicine
    Duke Global Health Institute at Duke University

    Mr. Denny is the Chief Operating Officer of the Duke Human Vaccine Institute (DHVI) and a Professor of Medicine and a Member of the Duke Global Health Institute at Duke University. He is also the director, of the Immunology and Virology Quality assessment Center. As the Chief Operating Officer of the DHVI, Mr. Denny has administrative oversight of a research portfolio including discovery and translational programs. He has over 30 years of immunology and virology experience studying host defense mechanisms. As part of the HIV clinical trials program, he has served on numerous committees for the NIH over the last two decades. Previously, he served on an expert panel for the CDC helping to establish clinical laboratory guidelines for using T-cell immunophenotyping in patients with HIV disease. In 1997, he received an NIH HIV Innovative Vaccine Grant award to study a new method of vaccine delivery. He is the principal investigator of the NIH-NIAID Division of AIDS Immunology Quality Assurance (IQA) Program, NIH-NIAID DAIDS External Quality Assessment Proficiency Oversight Laboratory (EQAPOL), and NIH-NIAID DAIDS Non-Human Primate Virology Reference Laboratory. In 2002, he was a Robert Wood Johnson Health Policy Fellow and served on the US Senate HELP Committee, staffing a number of health and science legislative issues. Mr. Denny has authored or co-authored more than 150 peer-reviewed papers and served on the editorial board of Clinical and Diagnostic Laboratory Immunology, Communications in Cytometry, and Clinical and Applied Immunology Reviews.

    Webinar Summary

    The history of the development of the current regulatory environment for laboratory developed tests (e.g., home brews). There will also be an outline of legislative proposals that are being suggested by current members of the house and senate, which, if enacted, could change what researchers do significantly.

    Learning Objectives

    • Understand how legislation evolves and the process of enacting it.
    • Understand the development and impact of rules on how legislation becomes a working practice.
    • Understand the impact of new legislation and rules about how you practice laboratory medicine.
    • Develop an understanding on how to get involved.

    Who Should Attend

    Those responsible for developing or using LDTs, managing laboratories, and maintaining laboratory accreditations.

    CMLE Credit: 1.0

  • Contains 3 Component(s), Includes Credits Recorded On: 02/26/2019

    A CYTO U Webinar presented by Vera A. Tang, PhD Keywords: small particles, virometry, scatter

    About the Presenter

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    Vera A. Tang, PhD
    Operations Manager
    Flow Cytometry and Virometry Core Facility at the University of Ottawa

    Vera helped to establish the Flow Cytometry and Virometry Core Facility at the University of Ottawa in 2013 when she became the first operations manager of the facility. Through her role in the core facility, she has channeled her passion for acquiring and sharing knowledge into teaching and research. Aside from new user training, she has developed courses in flow cytometry for undergraduate and graduate programs at the Faculty of Medicine at the University of Ottawa. Her research involves the development of retroviruses for use as fluorescence standards, as well as developing protocols for analysis of viruses and EVs by small particle flow cytometry. This work has led to the formation of a company of which Vera is a founding member and CSO. Vera is an active member of the cytometry community. She was accepted into the ISAC Shared Resource Lab Emerging Leaders Program in 2018 and is also serves as the co-president of the Canadian Cytometry and Microscopy Association.

    Webinar Summary

    This webinar will introduce fluorescent retroviruses as potential fluorescence reference particles, as well as how they can be used in small particle flow cytometry. It will also discuss features of viruses that are unique from available small particle reference materials and showcase new projects underway for development of MESF viruses.

    Learning Objectives

    • Improve understanding of the challenges for small particle flow cytometry.
    • Emphasize the need for reference materials and reagent development in small particle flow cytometry.
    • Introduce flow cytometrists to flow virometry.

    Who Should Attend

    Anyone who has an interest in the analysis of small particles such as EVs and viruses.

    CMLE Credit: 1.0

  • Contains 3 Product(s)

    A three-part CYTO U Webinar series on Validation, the Key to Translatable Flow Cytometry Keywords: Longitudinal, IQ, OQ, PQ, ERF, Inter-instrument

    Instrument qualification and method validation are two of the pillars required for obtaining cytometry data that are reliable and suitable for decision making. In this three-part webinar series all aspects of validation for flow cytometry, from the instrument to the assay, will be covered.  

    Validation, the Key to Translatable Flow Cytometry, Part 1: Method Validation—Overview, Concepts

    Validation, the Key to Translatable Flow Cytometry, Part 2: Method Validation—Planning and Executing

    Validation, the Key to Translatable Flow Cytometry, Part 3: Instrument Qualification

  • Contains 4 Component(s), Includes Credits Recorded On: 10/29/2018

    A CYTO U Webinar presented by Cherie Green

    About the Presenter

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    Cherie Green
    Senior Scientific Manager
    Flow Cytometry Biomaker Group
    Genentech

    Cherie Green has been a member of the cytometry community for over 20 years working in hematopathology and biotech laboratories. She is passionate about development of robust biomarker assays to support all phases of drug development. Currently, she oversees the Flow Cytometry Biomarker Group in the Development Sciences department at Genentech, a member of Roche Group in San Francisco, CA. Her group is responsible for the development and validation of clinical biomarker assays in the areas of infectious, autoimmune, and oncology diseases. She has served as the co-chair of Flow Cytometry Action Committee of the American Association of Pharmaceutical Scientists (AAPS) and has co-authored many consensus/recommendation papers on topics specific to drug development such as instrument and assay validation, sample stability, and receptor occupancy. 

    Webinar Summary

    The foundation of good data starts with the instrument. While substantial effort is often invested in development and validation of analytical methods or analysis, instrument validation is often neglected. It is essential to apply the same analytical and scientific rigor to the platform generating the data. From initial optimization and characterization of performance to establishing QC systems to ensure longitudinal data comparability, instrument validation strategies are critical components of generating robust and reliable data. This is true for all laboratory environments but particularly relevant for regulated labs providing decision-enabling biomarkers. Generating quality data plays a critical role in bringing new therapeutic options to the medical community—drugs which eventually manifest as successful new treatments for those individuals afflicted with disease.

    Learning Objectives

    In this webinar, you will learn the basic principles of instrument validation. Validation of flow cytometers used in regulated environments provides assurance that the output generated on these instruments is reproducible and precise. The most relevant elements of instrument validation include testing to verify that an instrument is installed properly and performs as intended. This includes establishing controlled procedures for installation, maintenance, calibration, cross-instrument standardization, and longitudinal performance monitoring. This course will review basic concepts of instrument validation and provide examples of each step in the process that can be applied in your lab.

    Who Should Attend

    Anyone interested in getting robust and reliable flow cytometry data.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 10/25/2018

    A Cytometry Part A Spotlight CYTO U Webinar presented by Yu “Max” Qian, PhD Keywords: DAFi, Semi-automated, Auto-gating, K-means, Clustering, FlowGate

    About the Presenter

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    Yu "Max" Qian, PhD
    Assistant Professor of Informatics
    J. Craig Venter Institute


    Dr. Yu “Max” Qian is an assistant professor of informatics at the J. Craig Venter Institute (JCVI). Max was one of the original developers of the flow cytometry (FCM) component of ImmPort, the NIAID/DAIT-funded immunology database and analysis portal, where he developed the FLOCK clustering method for computational identification of cell populations from FCM data. He led or collaborated with other FCM bioinformatics researchers in development of data transformation methods, information standards, data models, and software systems, including FCSTrans, MIFlowCyt, FuGEFlow, and GenePattern FCM suite. Collaborating with researchers from several institutions, he has recently focused on design and implementation of a web-based computational infrastructure—FlowGate (flowgate.jcvi.org)—for supporting clinical and translational research through data-driven reproducible analysis of FCM experiment data. He has been customizing data analytical pipelines and performing computational analytics of FCM data for multiple NIH-funded research projects, including the Respiratory Pathogens Research Center (RPRC) at University of Rochester and the Human Immunology Project Consortium (HIPC) center at the La Jolla Institute for Allery and Immunology.

    Webinar Summary

    Although auto-gating approaches have advantages over traditional manual gating analysis, there exist roadblocks before a cytometry lab can adopt an auto-gating approach for cell population identification in routine use. It was found that combining recursive data filtering and clustering with constraints converted from the user manual gating strategy can effectively address these roadblocks. This new approach is named DAFi: Directed Automated Filtering and Identification of cell populations. Design of DAFi preserves the data-driven characteristics of unsupervised clustering for identifying novel cell subsets, but also makes the results interpretable to experimental scientists through mapping and merging the multidimensional data clusters into the user-defined two-dimensional gating hierarchy. The recursive data filtering process in DAFi helped identify small data clusters which are otherwise difficult to resolve by a single run of the data clustering method due to the statistical interference of the irrelevant major clusters. Our experiment results showed that the results of DAFi, while being consistent with those by expert-centralized manual gating, have smaller technical variances across samples than those from individual manual gating analysis and the nonrecursive data clustering analysis. Compared with manual gating segregation, DAFi-identified cell populations avoided the abrupt cut-offs on the boundaries. DAFi has been implemented to be used with multiple data clustering methods including K-means, FLOCK, FlowSOM, and the ClusterR package. For cell population identification, DAFi supports multiple options including clustering, bisecting, slope-based gating, and reversed filtering to meet various auto-gating needs from different scientific use cases.

    Learning Objectives

    • Gain an understanding of what the cutting-edge auto-gating approaches can do and their limitations in general.
    • Learn how DAFi works, what it can do and cannot do, as well as how to apply DAFi to the analysis of polychromatic FCM datasets.
    • Assess the performance of an auto-gating approach using visualization and other computational methods.
    • Get to know the FlowGate cyberinfrastructure being developed.

    Who Should Attend

    Everyone who is interested in FCM bioinformatics, especially those who have been planning to apply auto-gating approaches for computational identification of cell populations from polychromatic FCM data.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 10/15/2018

    A CYTO U Webinar presented by Jennifer Wilshire, PhD & Tomas Baumgartner Keywords: panel design, spillover spreading, spillover spreading matrix, resolution impact matrix, sentinel panel design, brightness, compensation, controls, spectra

    The Presenters

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    Jennifer Wilshire, PhD
    Assistant Manager
    Memorial Sloan Kettering Cancer Center

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    Tomas Baumgartner
    Manager
    Flow Cytometry Core Facility
    Weill Cornell Medicine

    Webinar Description

    This webinar will focus on the process of building multicolor panels. We will break the process down into step-by-step guides based on the number of colors in your panel. This webinar will also provide a framework for those teaching multicolor panel design in an educational setting such as a Shared Resource Lab.

    Learning Objectives

    • Know the three processes used to build multicolor panels.
    • Choose the correct step-by-step process based on the number of colors in the panel.
    • Interpret spillover spreading matrix/resolution impact matrix to determine which colors contribute spreading to others.
    • Practice building a multicolor panel using all the tricks and tips discussed in the webinar.

    Who Should Attend

    Anyone who is designing multicolor panels or teaching others how to design multiparameter panels.

    CMLE Credit: 1.0

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  • Contains 4 Component(s), Includes Credits Recorded On: 09/10/2018

    A CYTO U Webinar presented by Teri Oldaker Keywords: regulatory, SOP, sensitivity, precision, reproducibility, quality management

    About the Presenter

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    Teri Oldaker
    Clinical Laboratory Consultant

    Teri is a licensed clinical laboratory scientist and certified cytometrist with over 40 years clinical laboratory and 34 years flow cytometry experience in reference laboratory settings.  She is currently in a consultant role for a number of clinical laboratories. Her prior roles include director of flow cytometry at Genoptix, Neogenomics, Genzyme Genetics, and Nichols Institute. Teri has served on the board of the International Clinical Cytometry Society (ICCS) as secretary/treasurer and councilor, and is on the faculty of both the ICCS and Clinical Cytometry Education Network (CCEN) Flow Courses. She is a member of the following ICCS committees: Advocacy, Certification, and Quality and Standards Committees. She has authored four book chapters and over 40 publications in the field of flow cytometry.

    Webinar Description

    This webinar will review the specifics on how to conduct a validation. We will begin with an introduction of the various regulated environments and accreditation bodies requiring assay validation. Then we will discuss how to conduct the appropriate validation for each environment. The distinction between assay qualification and validation will be reviewed. Various strategies to designing method validation protocols will be discussed. These recommendations will be aligned with the upcoming Clinical Laboratories Standards Institute (CLSI) guidelines regarding the number of samples, number of analytical runs, data analysis, and acceptance criteria. Examples of a biomarker validation and a clinical laboratory validation will be presented. Lastly, an update on the progress of a regulatory guidance document for the validation of flow cytometric methods will be presented.

    Learning Objectives

    In the second webinar of the series, you will learn the how to convert the principles learned in the first webinar into practice in your own laboratory. We will describe what you actually need to do to validate a method.

    • An introduction to the different types of regulatory environments.
    • Gain an understanding of the difference between assay development, optimization, and validation.
    • Gain an understanding of the fit-for-purpose and context-of-use validation approaches.
    • Learn to prepare user-friendly documentation.

    Who Should Attend

    Anyone using flow cytometry who wants to generate high-quality data.

    CMLE Credit: 1.0

  • Contains 4 Component(s), Includes Credits Recorded On: 08/13/2018

    A CYTO U Webinar presented by Virginia Litwin, PhD Keywords: Biomarker, fit-for-purpose, specificity, sensitivity, precision, LLOQ, calibration standard

    About the Presenter

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    Virginia Litwin, PhD
    Vice President, Immunology
    Caprion Biosciences Inc.

    Virginia Litwin is a thought leader in analytical method validation for flow cytometry. Bringing “Cytometry from Bench-to-Bedside” has been the focus of her professional activities since 1999, when she started working in translational medicine at Bristol-Myers Squibb. She co-founded the Flow Cytometry Action Program Committee (APC) within the American Association of Pharmaceutical Scientists (AAPS). The Flow Cytometry APC published the first papers on flow cytometry method and instrument validation, which Virginia was invited to present at the FDA Workshop on Clinical Flow Cytometry in 2013. Virginia is the chair of the Document Development Committee for a new Clinical Laboratory Standards Institute (CLSI) Guideline, H62: Validation of Assays Performed by Flow Cytometry. She is a councilor for both the ISAC and the International Clinical Cytometry Society (ICCS). In addition, she serves on the ICCS Advocacy Committee, whose mission is to interface with regulatory agencies.

    Webinar Summary

    This webinar will include an introduction of the concepts of analytical method validation. For flow cytometric assays, validation carries unique requirements owing to the challenges associated with measuring cells, the lack of cellular reference materials, and the complexity of the instrumentation. The focus of this webinar will be on how to apply the basic validation principles to flow cytometry and why guidelines appropriate to other methodologies cannot fully be applied to flow cytometry. One of the most challenging and contentious, but also most important, aspects of cell-based assay validation surrounds the demonstration of method accuracy and linearity. These points will be addressed in depth.

    Learning Objectives

    In this webinar, you will learn the basic principles of analytic method validation. We will define each of the validation parameters, and how those parameters can be addressed in flow cytometry method validation. We will also discuss how to interpret the validation data once it is generated and how the validation data influences the final use of the data.

    • Gain an understanding of the purpose of method validation.
    • Learn the basic validation parameters.
    • Gain an understanding of specific validation considerations for flow cytometry.
    • Gain an understanding of how the type of data generated in the assay influences the design of the method validation.
    • An introduction to interpreting validation data.

    Who Should Attend

    Anyone using flow cytometry who wants to generate high quality data.

    CMLE Credit: 1.0