About the Speaker

David Hedley, MD - 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, served on 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.  

CMLE Credit: 1.0

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.

CMLE Credit: 1.0

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.

CMLE Credit: 1.0

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

CMLE Credit: 1.0

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.

Learning Objectives

Panel Design, Spreading Error, Unmixing, OMIP design

Who Should Attend

Everyone interested in high-dimensional flow cytometry panels and OMIP-102n

CMLE Credit: 1.0

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.

CMLE Credit: 1.0

About the Speakers

Kathy Daniels - Clinical Biomarker Flow Cytometry Lead, Sana Biotechnology, Inc 

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

CMLE Credit: 1.0

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

Who Should Attend: SRL Managers and Staff

CMLE Credit: 1.0

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. 

CMLE Credit: 1.0

About the Speaker

Kylie Price
Chief Technology Officer
New Castle University
Malaghan Institute of Medical Research

Kylie Price is the Chief Technology Officer at the Malaghan Institute of Medical Research with 20 years’ experience providing strategic, scientific, and operational direction in multidisciplinary environments.  Kylie has strong stakeholder management and engagement skills and has attracted more than $35m of philanthropic funding over the past 12 years, supporting the creation of a distinguished centre of research excellence, the Hugh Green Cytometry Centre (HGCC).  Kylie leads a team of twelve highly skilled staff scientists in providing support to over 90 scientists at the Malaghan Institute.  The HGCC provides access to cutting-edge technology platforms (including full spectrum flow cytometry, histology, bioimaging, data science, molecular biology and mRNA for pre-clinical vaccine development), and she advises multiple research groups and companies both nationally and abroad. Kylie also has a strong leadership track-record, organizing high-profile networking events, such as CYTOAsia Singapore 2017, and directing international organizations, such as the Australasian Cytometry Society of which she is former President (2015-2016).  Kylie is a two-time NZ Woman of Influence Awards (2014 and 2018) 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 (ISAC) Council (2018-2020) and is currently serving as Chair of the Governance Committee and Secretary of ISAC (2020-2024).  In April 2023 Kylie was selected to join the NZ OnBoard Program and is currently a board observer for Orbis Diagnostics.

Webinar Summary

This webinar will cover an overview of Kylie’s career path and provide examples of how she promotes equity, employee engagement and participation within her team.  She will cover the SRL Career Progression Framework she developed with others at the Malaghan Institute of Medical Research and discuss how she encourages the appropriate recognition of SRLs and SRL staff scientists in publication. There will be a Q+A session and we hope for lots of questions and engagement.

Learning Objectives

1. New ideas for how to increase SRL staff engagement and participation 
2. A guideline to follow for the appropriate recognition of SRLs and SRL staff scientists in publication
3. An example of a non-traditional career path to Chief Technology Officer
4. Strategies for promoting equity in the workplace

Who Should Attend: Scientists who manage teams (PIs, researchers, SRL managers), SRL staff scientists, women in STEM, scientists interested in topics related to equity and inclusion.

CMLE Credit: 1.0

Veronica Nash, PhD
US Regional Head of Flow Cytometry
Cerba Research

Veronica earned her B.Sc and M.Sc in Medical Biotechnologies from “Tor Vergata” and from “Sapienza” University, respectively, in Rome, Italy. She continued her training at Ghent University, Belgium where she received her PhD in Medical Sciences. After her PhD, she joined Icahn School of Medicine at Mount Sinai in NYC where she leveraged her knowledge in virology and immunology with her Flow Cytometry expertise to identify host cofactors in the immune cell which are involved in HIV and HTLV infection. Afterwards, she moved to Indiana University in Bloomington, IN, and as a staff scientist she optimized a Flow Cytometry method to identify protein mutants to be used as a target in precision medicine for Cystic Fibrosis. Veronica’s career in industry started at Wuxi Apptec where she developed Flow Cytometry assays for QC of drug therapy products, such as CAR-T cells and TILs. Then she joined Labcorp Drug development (Covance) as a staff scientist where she focused on Flow cytometry assay validation, data review and instrument implementation. Currently, as the US Regional Head of Flow Cytometry department at Cerba Research, she is the scientific lead for high-parameters Flow Cytometry; in the last year, she led a global team through the implementation and harmonization of Cytek Aurora instruments across multiple sites. Veronica is also an active member of ISAC and ICCS; currently she is the leader of a working group which aims at promoting CLSI H62 guidelines education within the Flow Cytometry community.

Tania Nevers
Senior Principal Scientist
Bristol Myers and Squibb

Tania earned her B.Sc and M.Sc in Biology from St John’s University, Queens NY before receiving her PhD in Biomedical Sciences from Brown University in Providence, RI. Following her Ph.D., Tania pursued a postdoctoral training at Tuft’s University where her work combined areas of immunology, vascular biology and cardiac physiology to study several aspects of adaptive immunity in diverse inflammatory settings. She then moved on to a Contract Research Organization, Flowmetric Inc, where she served as a  Scientist and then Project Manager. Her primary responsibility was to design, review and execute complex flow cytometry experiments in both preclinical and clinical studies for client sponsored projects. She then transitioned to the large Pharmaceutical Industry where she currently resides as a Senior Principal Scientist at Bristol Myers and Squibb. She primarily works on high order assay development, validation and implementation, and is the subject matter expert for Cytek Aurora implementation across BMS, including integration and harmonization activities.   

When not at work, Tania enjoys spending quality time with family as well as listening to music, dancing and gardening. 

About the webinar:

The use of Spectral Flow Cytometry allows for a deeper characterization of immune subsets (immune profiling) than conventional flow cytometry and is more valuable with limited patient samples in biomarker discovery in drug development. This webinar describes how to implement and standardize Cytek Aurora instruments and how to design, develop, and validate 30+ parameters panels that can be successfully utilized in clinical trials.

Learning Objectives:

• Learn how to implement and standardize Cytek Aurora instruments
• Understand how to validate an assay for 30+ colors on a spectral instrument to use in clinical trials
• Learn solutions on challenges such as reference controls, unmixing strategies and experimental sept up when implementing a high-parameter assay for global clinical trials 

CMLE Credit: 1.0

Evan Jellison
Associate Professor & Director of Flow Cytometry
UCONN School of Medicine

Kristen Reifel
Vaccine Research Center
NIAID/NIH

Benjamin Fontes, MPH, CBSP
Biosafety Officer
Yale University Environmental Health & Safety

Avrill Aspland
Operations Coordinator
Sydney Cytometry

About the webinar:

This webinar will be an open discussion with members of the ISAC Biosafety Committee to share experiences of biosafety and how we work to seek continual improvements in our shared resource laboratories.  We will discuss the strategies we use to identify priorities, initiate improvements, and embed new ways of doing things.  We will also respond to biosafety-related questions submitted through a pre-webinar survey and to questions and comments submitted in real time through a live Q&A. Please complete the pre-webinar survey sent to your email!

CMLE Credit: 1.0

The Journey of Achieving High Standards in Multiparametric Flow Cytometry Data By Maria Jaimes

About the Presenter

Maria Jaimes
VP Scientific Commercialization
Cytek Biosciences 

Dr. Maria Jaimes earned her MD degree at the Universidad Javeriana in Colombia (South America). Dr. Jaimes completed her postdoctoral training at Stanford University in the department of Microbiology and Immunology. During her postdoc, she worked at characterizing the immune responses to both rotavirus and influenza viruses after natural infection and immunization. In 2005, Dr. Jaimes joined BD Biosciences.  While at BD, Maria worked in different aspects of quality assurance and standardization of flow cytometry assays. Since 2015, Maria has been working at Cytek Biosciences and is part of the R&D team who developed the Aurora Full Spectrum Cytometer. Dr. Jaimes 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

Dr. Jaimes will discuss her career in flow cytometry with emphasis on her work around standardization of flow cytometry assays and development of multicolor assays.  Dr. Jaimes will also discuss her journey of applying her knowledge in multicolor flow cytometry into the development of a new technology while working at a start-up, with the motivation of contributing to the generation of high-quality data. Maria will discuss the challenges faced by those dedicated to disseminating best practices in flow cytometry and the role that both academia and industry should play at enforcing those.

CMLE Credit: 1.0

About the Speaker

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

A Basic Introduction to the Science of Cell Sorting By Matthew Goff

About the Presenter

Matthew Goff
Senior Product Manager, Flow Cytometry
Beckman Coulter Life Science

Matthew began his career in flow cytometry doing graduate research at Virginia Tech followed by his role as a core lab manager at Eastern Virginia Medical School. After leaving EVMS for industry, Matthew stayed close to flow cytometry supporting researchers and industrial scientists in flow cytometry with reagents, software, and hardware products. In 2021 he became a commercial product manager at Beckman Coulter Life Sciences where his work is focused on sustaining the CytoFLEX Platform and launching new products. 

Webinar Summary
This webinar introduces scientists with an interest in learning more about flow cytometry an introduction to the mechanics of cell sorting, best practices when sorting cells and contemporary discussion topic associated with cell sorting.

CMLE Credit: 1.0

Should CYTO Women Go with the Flow? By Virginia Litwin

About the Presenter

Virginia Litwin
Scientist
Charles River Labs

Virginia Litwin is a thought-leader in validation and standardization 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 Community within the American Association of Pharmaceutical Scientists (AAPS). This group published the first papers on flow cytometry method and instrument validation. She was a counselor for the International Society for the Advancement of Cytometry (ISAC) and the International Clinical Cytometry Society (ICCS). She serves on the ICCS Advocacy Committee whose mission is to interface with regulatory agencies and has been an invited speaker at FDA/NIST on many occasions.

Virginia is the chair of the Document Development Committee for the guidance document, CLSI H62- The Validation of Assays Performed by Flow Cytometry. She edited the book, Flow Cytometry in Drug Discovery and Development, as well as journal special issues: JIM- Flow Cytometry Biomarkers and Translational Medicine (2011); Cytometry Part B- Receptor Occupancy (2015); Cytometry Part B- Cytometry Advancing Next Generation Drug Development (2021).

After obtaining a Ph.D. in Virology/Immunology from the University of Iowa, Virginia joined Lewis Lanier at DNAX as a post-doctoral fellow where she identified the KIR receptor, KIR3DL1 (CD158E1). She has held leadership roles in several contract research organizations. Currently she is a Research Scientist at Charles River Laboratories in Québec, Canada.

Webinar Summary

This webinar will review a wide variety of topics which impact the careers of CYTO Women. The importance of both being a Mentor and a Mentee will be reviewed. Case studies of how Sponsorship can impact an individual career and CYTO Women in general will be presented. Lastly, we will discuss how and when to change the things we cannot accept. Most of the webinar will be Q&A and hopefully a very lively discussion.

CMLE Credit: 1.0

Radioactivity and Flow Cytometry by Dagna Sheerar & Kathryn Fox

About the Presenter

Dagna Sheerar
Flow Cytometry Director
University of Wisconsin - Madison
Carbone Cancer Center Flow Cytometry Laboratory

Dagna Sheerar, SCYM(ASCP)^CM, has been working in flow cytometry shared resource laboratories for over 20 years and is currently the director of the Flow Cytometry Laboratory at the University of Wisconsin - Madison Carbone Cancer Center (UWFlow).  UWFlow is a busy core with a staff of 7 full time flow cytometrists, including Dagna, with 2 core laboratory locations on campus.  Dagna’s areas of interest are core administration and management, rigor and reproducibility, and user education. Since becoming 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 and currently serves on their Board of Directors. 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 recently completed the 45^th Annual Course 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.  

Kathryn Fox
Flow Lab Technical Manager
University of Wisconsin
Carbone Cancer Center Flow Cytometry Laboratory

Kathryn Fox, SCYM(ASCP)CM, is currently the technical manager of the Flow Cytometry Laboratory at the University of Wisconsin - Madison Carbone Cancer Center (UWFlow). She has been working for the UW Carbone Cancer Center Flow Cytometry Core Laboratory since 2015. Her flow cytometry interests include digging around inside instruments, any application that involves staining DNA, and being surprised by interesting new projects brought forward by the core’s diverse user base. Kathryn graduated from Iowa State University with a BS in Biology and then from the University of Wisconsin – Madison with a PhD in Cancer Biology. She was first introduced to flow cytometry in graduate school, but her main focus at that time was fluorescence microscopy. After graduate school, she worked as an applications scientist in multiphoton microscopy. Since diving into flow cytometry, Kathryn has been an active participant in GLIIFCA and the Annual Course in Cytometry, and finally got to attend her first in-person CYTO in 2022. 

Webinar Summary

This seminar will focus on how a flow core can respond to the needs of researchers to analyze radiotherapy treated samples in real time via flow cytometry.  We will present our process for providing this service from the initial idea, partnering with our Small Animal Imaging and Radiotherapy Facility, working with our institutional Radiation Safety group to test various cleaning methods for removing radioactive particles from the instrument post-analysis, instrument modifications, working with the original equipment manufacturer to ensure compliance with our existing service contract and safety for the engineers servicing the equipment, and additional safety measures and training procedures developed specific to this service.

CMLE Credit: 1.0

About the Speaker

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

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About the Speaker

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

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About the Speaker

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

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About the Speaker

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

About the Speakers

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.

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 45^th 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

About the Speakers

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.

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

About the Speaker

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

About the Speakers

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. 

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

About the Presenter

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.  

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

About the Speaker

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

About the Presenter

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.

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

About the Presenter

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

About the Presenter

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

• 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 

About the Presenter

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

• 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)

About the Presenters

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.

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

About the Presenter

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

About the Presenters

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.

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.

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

• 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

About the Presenters

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.

 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

About the Presenter

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

About the Presenter

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

About the Presenter

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

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.

About the Presenter

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

About the Presenter

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

About the Presenter

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

About the Presenter

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

• 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

About the Presenters

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.

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

About the Presenters

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.

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

About the Presenter

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