A CYTO U Webinar presented by David Hedley
Keywords: Intracellular antigens, Cell signaling, Leukemia, Cytokine
About the Speaker
David Hedley, Ph.D - Emeritus Professor of Medicine and Senior Scientist, University of Toronto and Princess Margaret Cancer Centre
Dr. David Hedley is a physician/scientist, trained in medical oncology and wet lab research at the Royal Marsden Hospital, UK 1975-81. Attending physician in medical oncology and laboratory scientist, Royal Prince Alfred Hospital/Ludwig Institute for Cancer Research, Assistant Professor of Medicine, University of Sydney, 1981-1989. Since 1990 he has held dual appointments at Princess Margaret Hospital, Toronto, as medical oncologist, specializing in breast and gastrointestinal cancers, and senior scientist with a laboratory program focused on cancer biology and experimental treatment. This laboratory program made extensive use of flow cytometry under my direct supervision, studying laboratory models of cancer, as well as interfacing to a large clinical trials program using novel analytical methods for patient monitoring.
Contributions to Flow Cytometry
Member of ISAC since 1982, serviedon Council 1989-1992, chair of Site Selection Committee, Education and Scientific Advisory Committees. Cytometry Editorial Board 1986-2024. ISAC Fulwyler Awardee 2014 for innovative excellence. Annual Flow Cytometry Methods Course faculty 1999-2019
Webinar Summary
This Webinar will discuss the application of flow cytometry to study dynamic processes that regulate cell function in health and disease. Major focus is on intracellular signaling and patient monitoring during treatment with targeted agents. Although emphasis is on cancer treatment, the Webinar also considers intracellular signaling pathways that regulate cytokine production in response to infectious agents. I will outline technical aspects of intracellular antigen measurements by flow cytometry, including a general approach to the study of dynamic post-translational modifications, experimental design, and integration into mechanism-based clinical trials of novel agents
Learning Objectives
1) How to do good quality intracellular staining while maintaining light scatter and surface antigens (not as difficult as you might think). 2) The huge range of antibodies to structural and functional intracellular antigens, most of which have never been tested in flow. Provided you have a clear question and trustworthy controls, they provide the opportunity for innovative research projects. 3) A general approach to the study of dynamic intracellular processes in live cells, using antibodies to posttranslational modifications such as phosphorylation. Emphasis on live cell function, including dose/response and time course studies. Analytical power when applied both to basic research projects, and to patient monitoring in clinical trials.
Who Should Attend
This webinar should be of broad interest both to the research and the clinical flow communities, especially the curiosity-driven, and to students and supervisors looking for innovative projects.
A CYTO U Webinar presented by Joshua Welsh and Vera Tang
Keywords: Calibration, Optimization, Small Particles, Quantitative Flow Cytometry, Standardization
the Speakers
Joshua Welsh- Staff Scientist, BD Sciences and Director, the Measuring Stick, Ltd
Joshua Welsh, Ph.D., recently joined BD’s Advanced Technology Group as a Staff Scientist where he works on next-generation standardization. He earned his BSc (Hons) in Physiology with Pharmacology from the University of Leicester and completed his Ph.D. in Translational Medicine at the University of Southampton. Dr. Welsh continued his training at the National Institutes of Health, where he advanced to a Staff Scientist role in Dr. Jennifer Jones' Translational Nanobiology Section. Dr. Welsh led initiatives like MISEV2023 and MIFlowCyt-EV, served as a member-at-large on the board of ISEV, chaired the ISEV reference materials task force, and led the ISEV-ISAC-ISTH EV flow cytometry working group. He is also the developer of FCMPASS, an optimization, calibration, and reporting tool for small particle flow cytometry.
Vera Tang - Manager & Adjunct Professor, University of Ottawa
Vera Tang, Ph.D., is an Adjunct Professor at the University of Ottawa in Canada. She has been managing the uOttawa Flow Cytometry and Virometry Core Facility since 2013, where she helps to educate the next generation of researchers and provides consultation and research support to both academia and industry. Her work focuses on developing reference materials, workflows, and best practices recommendations for small particle analysis. Vera is an ISAC SRL Emerging Leader (2018-2023), ISAC Councilor (2024-2028), Vice President and past President of the Canadian Cytometry & Microscopy Association, and an Associate Editor of Cytometry, Part A.
Webinar Summary
This workshop will build from basic principles of experimental setup to optimization and calibration of instruments, to providing resources and knowledge to perform advanced characterization methods for data analysis.
Learning Objectives
To gain understanding on how to characterize a flow cytometer’s potential for small particle detection.
Who Should Attend
This webinar is intended for all current and potential small particle enthusiasts (novice to expert) interested in performing small particle analysis.
A CYTO U Webinar presented by Jane Srivastava
Keywords: Equity, Training, Inclusivity
About the Speaker
Jane Srivastava, Core Director for Flow Cytometry, Light Microscopy and Histology, J. David Gladstone Institutes
Originally from Christchurch, New Zealand, Jane Srivastava (she/her) completed her undergraduate studies in plant and microbial biology at the University of Canterbury. She worked as a lab technician before discovering flow cytometry in 2003 and has been in the industry ever since, working as a flow lab manager at Imperial College London and designing practical and theoretical flow courses for undergraduate and postgraduate students. Jane is now director of the Flow Cytometry Core at the Gladstone Institutes and has been a mentor for Gladstone’s PUMAS (Promoting Underrepresented Minority Advancement in the Sciences) summer internship program. In 2024, Jane took over the directorship of the Light Microscopy and Histology at Gladstone Institutes. Jane considers herself a career technician and is passionate about bringing recognition and support to those working in a scientific technical career
Webinar Summary
Although education methodology has advanced considerably form even a few years ago, there is still a general need to include equitable teaching practices in training sessions offered by shared resource facilities. Jane will summarize and give examples of these practices, from how to introduce feedback literacy for both students and staff, to how to utilize technological platforms to enhance student learning.
Learning Objectives
Learn how to modify training to make it more inclusive. Understand how equitable training practices lead to less repetition of key concepts and reduces staff burden
Who Should Attend
Anyone that is offering training to people using an SRL.
A CYTO U Webinar presented by Adrian Liston
Keywords: FlowCodes, in vivo analysis, T regulatory cells
About the Speaker
Adrian Liston, PhD - Professor of Pathology, University of Cambridge
Dr. Liston has extensively worked in the cellular control over immune/tolerance switches, and how molecular defects in these switches can lead to pathologies ranging from autoimmunity and primary immunology to diabetes and neuropathology. By researching a broad range of pathologies, and using both patient samples and animal models, Liston’s research is able to identifying the common cellular pathways to pathology. Liston has been awarded the Francqui Chair, the Eppendorf Prize and fellowship to the Academy of Medical Sciences and the Royal Society of Biology. Liston currently serves as the Editor-in-Chief of Immunology & Cell Biology.
Webinar Summary
The tissues are the site of many of the most important immunological reactions, yet the immunology of the tissues has remained relatively opaque. Recent studies have identified Foxp3+ regulatory T cells(Tregs) in several non-lymphoid tissues. These tissue-resident populations have been ascribed unique characteristics based on comparisons to lymphoid Tregs. To understand key aspects of this tissue Treg population, we need technological approaches thatare suitable for the assessment of low cell number sources with high dimensionality, such as flow cytometry, to be coupled with massively parallel screening technologies (e.g. CRISPR screens, TCR clonality screens). We modified the ProCodes technology into FlowCodes, a flow cytometry-based platform suitable for in vivo analysis. Using this system, we found that T cell receptors (TCRs) extracted from tissue Tregs conferred Treg fate and also multi-organ homing. These results demonstrate that tissue-resident Tregs are largely constituted by broadly self-reactive Tregs, characterized by transient multi-tissue migration and a common residency program.
Learning Objectives
Regulation of T cell mediated tolerance Understanding tissue resident versus lymphoid origin regulatory T cells Development and Utilization of FlowCodes: A flow cytometry-based platform suitable for in vivo analysis
Who Should Attend?
General interest to beginner and advanced flow cytometrists SRL staff interested in development and implementation of multiplexing technology Immunologists interested in T cell biology
A CYTO U Webinar presented by Florian Mair
Keywords: Education, Panel Design, 50-Color, Training
About the Speakers
Florian Mair - Scientific Director of Flow Cytometry Core Facility, ETH Zurich, Switzerland
Florian is currently working as the Scientific Director of the Flow Cytometry Core Facility and Senior Scientist Immunology at ETH Zurich, Switzerland. After graduating with an MSc in Molecular Biology from the University of Vienna, he received his PhD in Molecular Life Sciences from the University of Zurich in 2014, and spent several years at the Fred Hutchinson Cancer Center in Seattle, USA. During the past decade, he has been involved extensively with different high-dimensional cytometry platforms as well as scRNA-sequencing techniques, including multi-omic methods. His main scientific interests are the function of dendritic cells and T cells in non-lymphoid tissues during health and disease, teaching best practices in cytometry and developing new flow cytometry panels and approaches.
Webinar Summary
To understand the function of the human immune system it is imperative to capture as much information as possible from often size-limited human samples. To do so, we have developed the first 50-color spectral flow cytometry panel to comprehensively study the functional state of the human immune system in PBMCs and tissue samples. The panel contains lineage markers for all major immune cell subsets, and an extensive set of phenotyping markers focused on the activation and differentiation status of the T cell and dendritic cell (DC) compartment. The panel has also been tested to be suitable for cell sorting, which allows very fine-grained isolation of immune subsets utilizing 50 markers at the same time.
To establish such a complex panel, we utilized a new metric termed unmixing spreading error, that evaluates the fluorochrome-specific increase in background that is inherently generated when unmixing highly complex panels. In this presentation we show the systematic workflow we used to develop the panel, how we evaluated panel performance, and how OMIP-102 was developed to its final state.
A CYTO U Webinar presented by Mariela Bollati-Fogolin and Ziv Porat
Keywords: SRL – End Users – Training
the Speakers
Ziv Porat - Head of Flow Cytometry Unit, Life Sciences Core Facilities, Weizmann Institute of Science, Israel
Ziv Porat obtained his PhD and post doctorate studies in Cell Biology from the Weizmann Institute of Science, Israel. Since 2011 he is part of the Weizmann Institute Flow Cytometry unit, and serves as head of the unit since 2018 as a Senior Staff Scientist. He specializes in Imaging Flow Cytometry, and participated in more than 500 different projects utilizing this technology. He took part in more than 100 publications in this field, including development of novel Imaging Flow Cytometry applications. He participates and lectures in numerous international conferences, including the CYTO meetings since 2015, and he is part of the ISAC emerging leaders program (2019-2024). Dr. Porat is the organizer of the Israeli annual Imaging Flow Cytometry meetings since 2012. He is the co-founder of the Flow Cytometry Society of Israel (FCSI) and organizer of the Israeli flow cytometry conferences and workshops.
Mariela Bollati - Head Cell Biology Unit, Institut Pasteur de Montevideo, Uruguay
Since 2006, Dr Mariela Bollati-Fogolín has been working at the Institut Pasteur de Montevideo, Uruguay, as Head of the Cell Biology Unit (CBU). There, she has been instrumental in establishing a vibrant and internationally recognized flow cytometry and cell culture facility, one of the first of its kind in South America. Managing the CBU Shared Resource Laboratory and its staff, she provides support and assistance to research scientists and companies throughout the Latin American region. She has actively participated in organizing several hands-on flow cytometry workshops. She has served on the Council (2020-2023) of the Uruguayan-Argentine Flow Cytometry Society (GRCF). At present, she is the ISAC CYTO Women Taskforce chair and serves as the Uruguayan Deputy Director of the Latin American Center for Biotechnology
Webinar Summary
Drs. Mariela Bollati-Fogolín and Ziv Porat will co-present the upcoming webinar about developing and delivering an effective training program for Shared Resource Laboratories. This webinar will focus on how to teach users how to utilize flow cytometry equipment properly. By examining real-world case studies, the presenters will illustrate the potential pitfalls of allowing inexperienced individuals unsupervised access to cytometry equipment. Additionally, they will emphasize the critical role of a quality management system (QMS) in ensuring the success of any internal training program.
This webinar is the second installment in a three-part series dedicated to training resources. Last month's webinar, presented by Kathy Daniels and Derek Davies, explored external training solutions. The series will conclude in December with a presentation by Jane Srivastava on leveraging modern educational theories to create equitable training programs.
Learning Objectives
In this webinar the attendees will learn: 1- Two case studies to deliver an Internal Training Program 2- The importance of a good training and education for end users 3- The relevance of having a quality management system (QMS) for Internal Training Program
Who Should Attend
SRL managers, staff and users of shared resource laboratories. Users of flow cytometry technologies or anyone who is interested in flow cytometry education.
Kathy currently works to provide support on flow cytometry experimental design and oversight for global clinical trials. She also serves on the Steering Committee for the MetroFlow regional cytometry group and volunteers within multiple committees for the International Society for the Advancement of Cytometry. In the past she has worked in both pharmaceutical and academic settings in the capacity of a flow cytometry shared resource facility where she supported end users in experimental design, education, experimental education and interpretation of data. Prior employees include The Whitehead Institute for Biomedical Research, Memorial Sloan Kettering Cancer Center and Regeneron Pharmaceuticals.
Derek Davies - Cytometry Trainer, Educator and Consultant, Derek Davies Cytometry Derek has been involved in Cytometry for over 40 years. He first used cytometry to measure DNA content in cervical cells but quickly realised the power of the technology. For 25 years he ran a Flow Cytometry SRL at the London Research Institute and then the Francis Crick Institute in London, UK. In that time he trained thousands of users and also developed the careers of SRL staff. In 2018, he became the Science Technology Platform Training Lead at the Francis Crick Institute where he developed training for users external to the Institute. In 2023, he transitioned to semi-retirement but is still active in education, training and consultancy. He is current Chair of flowcytometryUK, sits on the Cytometry Committee of the Royal Microscopical Society and is a former ISAC Councilor.
Webinar Summary • Introductions • Start w/ importance of training - rigor and reproducibility • Responsibilities of the SRL in training and education • Bottlenecks and challenges faced when handling training internally at an SRL • What are the outsourcing options? • Theory, practical, in person, virtual, core curriculum • Vendor based training (on site vs virtual) • Free & Paid Training Resources (benefits vs limitations) • Annual Training Courses Examples • How to evaluate these training courses? • Metrices for evaluation • How can this be done before you commit to one (or more) solutions?
Learning Objectives In this webinar we hope to have everyone leave with a better understanding of why training and education is so critical and how to evaluate training opportunities that are available outside the SRL. As part of this, we will share what some options are, and how to evaluate them and their effectiveness.
Who Should Attend
SRL managers, staff and users of shared resource laboratories. Users of flow cytometry technologies or anyone who is interested in flow cytometry education
Why can't they play nicely? Navigating unexpected challenges in designing new flow cytometry panels and expanding existing ones - Anna C. Belkina, MD, PhD & Kelly Lundsten
Think Like an Engineer – Early Detection and Troubleshooting of Cytometer Problems - Rachael Sheridan, PhD SCYM (ASCP)CM & Matthew Goff
A Beginner's Guide to Computational Cytometry: From Algorithms to Applications - Sofie Van Gassen
Label-Free Imaging and High Dimensional Analysis - Peter O'Toole, PhD & Karen Hogg, PhD
Assay Design and Standardization: What Can Researchers Learn From Clinical and Vice Versa? - Lyana Setiawan, MD, PhD, Paul Hutchinson, Paresh Jain
Image Cytometry Analysis Using AI Techniques - Aida Meghraoui, PhD, PharmD & Polat Goktas, PhD
A CYTO U Webinar presented by David Leclerc
Keywords: SRL Promotion, Marketing Approaches, Communication with Users, Optimizing the Value of SRL Services
About the Speaker
David Leclerc
Director, CAT Facility at the University of Chicago
David Leclerc is the Director of the Cytometry and Antibody Technology (CAT) Facility at the University of Chicago. With over 20 years of expertise in the field, David is passionate about implementing cutting-edge technologies and fostering innovative multi-media solutions to encourage its adoption within the research community. Beyond his role at the University of Chicago, David is a tenured member of the GLIIFCA Board of Directors, President of the Chicago User Group, Creator of the FixYourFlowDepot (a video repository on cytometry maintenance and repair), and host of the ChUG Podcast.
Webinar Summary
An important function of a shared service laboratory (SRL) is the promotion of novel technologies for the benefit of the research community. This webinar introduces a systematic method to help SRL’s optimize their efforts when advertising new instruments/services within their facility. The GSTIC approach (Goal/Strategy/Tactics/Implementation/Control) allows SRLs to design a methodical approach to identify objectives/tactics, and most importantly, a clear set of expectations to evaluate the success of your strategies.
Learning Objectives
1. Understanding the GSTIC Approach: Participants will gain knowledge about the GSTIC approach and its application in increasing the value of services within a Shared Service Laboratory (SRL).
2. Applying GSTIC for Marketing Goals: Participants will learn how different components of the daily activities within an SRL can be utilized to support marketing objectives, emphasizing practical application.
3. Exploring Communication Channels: Participants will gain insights into the various communication channels that exist between an SRL and its users. This includes an examination of the pros and cons associated with different options, fostering a comprehensive understanding of effective communication strategies within an SRL context
A CYTO U Webinar presented by Caroline E. Roe, MLI
Keywords: High dimensional cytometry, Spectral flow, Mass cytometry
About the Speaker
Caroline E. Roe, MLI Managing Director, Cancer and Immunology Core Vanderbilt University, Nashville, TN, USA
Ms. Roe received her B.S. in Genetics, Cell Biology, and Development from the University of Minnesota-Twin Cities and Master of Laboratory Investigation from Vanderbilt University in Nashville, Tennessee, where she has managed the Mass Cytometry Center of Excellence since its inception in 2017. As director of the Cancer and Immunology Core at Vanderbilt, her focus is on increasing both the standardization and accessibility of cutting-edge cytometry techniques for application across a broad range of basic and translational research projects.
Webinar Summary This webinar aims to give an experience-driven overview of considerations for choosing the appropriate high dimensional flow cytometry technology for a project as well as guidance for those who have experience in one technology and are transitioning to the other.
Learning Objectives Attendees will leave the webinar with a strong foundation in high dimensional cytometry principles and applications, as well as sufficient working knowledge to select the appropriate technology for their study.
Who Should Attend: Users who are interested in high-dimensional cytometry of any kind.
