As always, GLIIFCA is planning several Lunchtime Roundtables to provide the opportunity for open discussion with the experts. The following roundtables are currently planned:
Antibody Rigor and Reproducibility
Maurice Shen, Head of Academic Relations, BenchSci
Antibodies play an essential role in every flow cytometry experiment. However, sourcing reliable antibodies and achieving reproducibility has been documented as a major challenge due to various factors. In this roundtable, we will discuss what is known about the “Antibody Crisis” and established antibody validation methods. We’ll then open the discussion to the audience to share their experience on the best practices for antibody selection for flow experiments, and discuss potential solutions to this issue.
Instrument Standardization: Who, What, When, Where, Why, & How
Dagna Sheerar, Manager, UWCCC Flow Lab
A lively and interactive discussion concerning the importance of instrument standardization in the context of rigor and reproducibility, including, but not limited to; examples of how to incorporate standardization techniques into your Quality Control routines, experimental data acquisition, user training and education, and the importance of standardization in the use of computational/machine learning data analysis algorithms.
Analysis of Viruses and Virus-like Particles by Flow Cytometry
Vera A. Tang, Operations Manager & Adjunct Professor, uOttawa Flow Cytometry & Virometry Core Facility, Ottawa, ON, Canada.
Advances in the technology of cytometers now allow for some commercial instruments to detect biological particles (such as viruses and EVs) down to the 100 nm diameter range with minor to no modifications to default instrument configurations. For virologists, this opens a new area of study where they can now perform phenotypic analysis on viruses and virus-like particles on a single-particle level. However, since these particles are very close to the limit of detection for current cytometers, and cytometers differ in optical configurations, it is very important that methods and reference materials are used for standardized data reporting. For this session, we will cover the following topics:
- Reference Materials for QC and standardization for small particle FCM
- Methods for standardized data reporting for small particle FCM fluorescence and scatter calibration
- Sample preparation and controls for small particle FCM analysis
- Challenges to distinguishing viruses, virus-like particles, extracellular vesicles
Considerations for High-Dimensional Panel Design
Derek Jones, Technical Director for Research and Development, University of Pennsylvania
As newer flow cytometers support an expanding number of fluorescent parameters, the need for rational panel design becomes increasingly important. We will discuss strategies for the design of high-dimensional flow cytometry panels, including fluorochrome brightness, antigen density and co-expression, compensation, and spreading error.
Imaging Flow Cytometry: Emerging Applications and Analysis Tools.
Robert Thacker, Luminex
Come join in on a discussion that will cover some of the latest applications, get your questions answered about current applications, and learn about the latest features and masks in IDEAS designed to enhance the depth of your data analysis.
Young GLIIFCA: What Can GLIIFCA And ISAC Do For My Career?
Jessica Back, Associate Director, Microscopy, Imaging, and Cytometry Resources Core, Karmanos Cancer Institute, Wayne State University.
There are many opportunities for professional development available through GLIIFCA and ISAC, but you may not know what these programs are or how to access them. This Roundtable will discuss the volunteer opportunities and travel/poster awards available through GLIIFCA; ISAC leadership development programs; ISAC taskforces and committees; CytoU offerings and opportunities for participation; and the SCYM Exam. Find out how you can get involved and what GLIIFCA and ISAC can do for you.
Minimal (measurable) Residual Disease – steps to implementation
Michael Keeney, Associate Scientist, Lawson Health Research Institute, London, Ontario, Canada
As new drug therapies are increasingly relying on MRD as a surrogate endpoint for treatment effectiveness, demands on clinical labs to introduce methodologies that were once performed only in research or highly specialized laboratories are increasing. Using MRD as an example, this roundtable discussion will focus on appropriate instrument set up, sample handling, and statistical considerations needed for the successful implementation of clinical and translational research. Particular focus will be placed on rare event analysis. This roundtable will provide a forum for participants to discuss issues in the design and implementation of these techniques.
Cell Sorting: Setting Yourself Up for Success
Lauren Nettenstrom, MS, SCYM(ASCP), Instrumentation Technologist, UWCCC Flow Cytometry Laboratory
We will discuss the myriad steps you need to take well before you get to the instrument to set yourself up for a successful sort. Topics will include knowing your sorter, biosafety considerations, optimal panel design, and tips for sample prep.
Education in Flow and Image Cytometry
Dr. Alexis Conway, Graduate Course Director/Coordinator and Flow Cytometry Specialist, Roswell Park Comprehensive Cancer Center, Buffalo, New York
This round table is for engaging ideas for cytometry education. We will discuss development of materials for graduate students and core facility users; as well as outreach efforts for educating people in flow and image cytometry. Cyto Youth and other cytometry education initiatives will be discussed.
MiSet RFC Standards: defining a universal minimum set of standards required for reproducibility and rigor in research flow cytometry experiments
Fabienne Lucas, MD PhD, Clinical Pathology Resident Brigham and Women’s Hospital Boston/ Clinical Fellow in Pathology Harvard Medical School, ISAC Marylou Ingram Scholar 2019-2024.
Important aspects of instrumentation, experimental design, data analysis and data reporting to produce valid flow cytometry data are widely available in the literature. Comprehensive guides covering the principles, techniques and applications of flow cytometry used in specific biomedical fields have also been published. However, there is currently no consensus on the minimum set of standards that overall define a rigorous and robust flow cytometry experiment in translational biomedical research. MISet RFC Standards (Minimum Set of Research Flow Cytometry Standards) is a new interdisciplinary initiative that will build on, integrate and simplify existing resources, with the mission to provide a single-resource broadly applicable guideline to increase the rigor and reproducibility of translational research flow cytometry experiments. Join this round table discussion to learn more about the aims and anticipated timeline of this project, and to discuss strategies for interdisciplinary implementation.
High Dimensional Data Analysis
Anna Belkina, Assistant Professor of Pathology and Laboratory Medicine, Associate Director, Flow Cytometry Core Facility, Boston University School of Medicine