How differences in detecting photons and ions impact experimental considerations in cytometry
In traditional fluorescence flow cytometry (FFC), the reporter used is a fluorescent molecule that is measured using a photomultiplier tube (PMT) or more recently with avalanche photodiodes (APDs). Mass cytometry, or CyTOF® technology, is a relatively new single-cell analysis platform with which cellular targets are labeled with metal-tagged antibodies and detected and quantified by time-of-flight mass spectrometry.
For each method, specific issues related to the detection system must be considered during this process.
From this webinar, you will learn how to improve panel design for both FFC and mass cytometry by understanding:
Brightness and sensitivity as a measure of reporter intensity
The impact of target abundance when pairing antigen and reporter
How to factor in cost and sample recovery when choosing between these two techniques
The true impact of acquisition speed on data quality
The similarities in data analysis and application of proper controls
About the presenter:
Tim Bushnell PhD, MBA Co-Founder Expert Cytometry
Contact us to learn more about mass cytometry.
For Research Use Only. Not for use in diagnostic procedures.
Videos and Webinars
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Watch on demand
Check out this library of recorded seminars, webinars and other videos available for you to learn more about Fluidigm and how CyTOF® and microfluidics technologies can accelerate your research.
Learn why CyTOF technology and microfluidics has played a significant role in the methods, applications and findings from experts in the fields of immunology, cancer immunotherapy, drug discovery, vaccine development and infectious disease.
Uncovering Immunological Mechanisms of Protection from Infection and Vaccination
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WEBINAR
Informing vaccination strategies with identification of protective immunological features using mass cytometry
In this presentation, Marcelo Sztein, MD, shows examples of his group’s use of mass cytometry to help advance the fields of vaccine development and pathogen-host interactions. He provides an in-depth look at his studies of Salmonella typhi (S. typhi), the causative agent of typhoid fever, an infectious disease of great public health importance.
In this webinar you will learn how Sztein’s team successfully used mass cytometry to
identify the key role that regulatory T cells appear to play in the development of typhoid disease following an oral challenge with wild-type S. typhi in humans
investigate in detail the differences observed in T cell-mediated immunity elicited by oral immunization with the licensed Ty21a-attenuated typhoid fever vaccine in children and adults
uncover early cell type-specific epigenetic modifications elicited in human gut cells following ex vivo exposure to S. typhi, which are likely to influence downstream immune responses in the gut mucosa microenvironment.
The COntAGIouS trial is a collaboration led by the research university KU Leuven and University Hospital Leuven to perform an in-depth characterization of the dynamic host immune response to coronavirus SARS-CoV-2.
Immune monitoring studies, especially in the age of COVID-19 disease, require cytometry assays that
are quick to implement with an easy workflow
have a fixation step early in sample processing
are high-parameter to ensure the most information possible per cell
have a streamlined, reproducible and unbiased data analysis pipeline
have proven reproducibility when performed at multiple sites.
The Maxpar Direct Immune Profiling Assay, designed for use on Helios™, a CyTOF® system, meets all these criteria and helps De Smet’s team observe the different immune system players and their interactions in COVID-19 patient samples.
About the presenter:
Frederik De Smet, MSc, PhD Assistant Professor
University of Leuven, Belgium
Fill out the form below to watch this video.
For Research Use Only. Not for use in diagnostic procedures.
Live-cell barcoding with Cd-CD45 antibodies
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WEBINAR
Sample multiplexing with barcoding approaches
With CyTOF® technology, sample multiplexing is easily achieved with cell barcoding approaches that label individual samples before staining and acquisition of the combined samples. Barcoding reduces sample preparation time while increasing experimental throughput and improving data consistency and quality.
Several options for cell barcoding are available including live-cell barcoding, which labels human white blood cell samples with a unique 3-digit isotope code, using cadmium-labeled anti-CD45 (a pan-leukocyte marker) antibodies.
In this recording you will learn:
Why barcoding is advantageous to your experimental design
The products offered by Fluidigm for this application
The step-by-step method including data analysis
Workflow tips and tricks
About the presenter:
Michelle Poulin, PhD Manager, Proteomics Field Applications
Fluidigm
This webinar first aired on September 23, 2020, at the 9th Annual Fluidigm Mass Cytometry Summit: Virtual event.
Contact us to learn more about mass cytometry and Cd-CD45 antibodies for use in live-cell barcoding.
For Research Use Only. Not for use in diagnostic procedures.
Understanding the immunosuppressive functions of cancer-associated fibroblasts (CAFs)
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WEBINAR
Understanding the immunosuppressive functions of cancer-associated fibroblasts (CAFs) in lung cancer
Hear how Merck Senior Scientist Handan Xiang, PhD, used Imaging Mass Cytometry™ (IMC™) to investigate the spatial relationship and functional role of CAFs with regard to their immunosuppressive effect and interactions with infiltrating immune cells in lung squamous cell carcinoma. In this presentation Xiang details her investigation of a mechanism that may underlie CAF impact on monocyte differentiation in lung squamous cell carcinoma.y.
About this webinar:
In this webinar presented initially at the Mass Cytometry Virtual Summit meeting, Xiang introduces her approach to answer key questions about the role of CAFs in lung cancer and discusses:
Potential mechanisms of CAFs in the stroma in resistance mechanisms of immune checkpoint blockade
How CAFs can impact monocytic migration and differentiation into myeloid-derived suppressor cells
Insights as to how NOS2 might serve as a potential drug target to restrain the immunosuppressive effect of CAF-educated cells
Xiang reveals how she used IMC in this biopharma project investigating a CAF role in a pathway of migration and differentiation to impact T cell expansion and function.
About the presenter:
Handan Xiang, PhD Senior Scientist
Merck Research Laboratories
Handan Xiang is a Senior Scientist in Discovery Immunology at Merck. She previously was an Associate Scientist and postdoctoral fellow in Discovery Oncology at Merck, where she completed the work presented in this webinar. She earned her PhD in the Department of Pharmacology and Cancer Biology at Duke University.
For Research Use Only. Not for use in diagnostic procedures.
Webinar | Comprehensive Landscape of the Tumor Microenvironment Analyzed
with CyTOF Technology
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WEBINAR
Hiroyoshi Nishikawa, MD, PhD, discusses Imaging Mass Cytometry in his presentation titled Comprehensive Landscape of the Tumor Microenvironment Analyzed with CyTOF Technology.
Immune escape mechanisms such as the induction or recruitment of immunosuppressive cells and the increased expression of various immunosuppressive molecules including PD-1/PD-1 ligands are essential processes during cancer development and progression, leading to the development of complex immune-suppressive networks in the tumor microenvironment. Thus, high-dimensional immune profiling using CyTOF® technology for single-cell phenotype and function analysis plays an important role in developing more effective cancer immunotherapeutic strategies and in defining biomarkers for stratifying responders and nonresponders via the detailed analysis of immune responses in cancer patients.
About this webinar:
In this presentation from the July 2020 Fluidigm Oncology and Infectious Disease Virtual Summit, Nishikawa outlines the current status of cancer immunotherapy and recent advances in the field. He also describes:
The importance of biomarkers in the cancer immunotherapy field and in PD-1 blockade therapy
His work in assessing a strong suppressive tumor microenvironment in EGFR-mutated non-small cell lung carcinoma (NSCLC) tumors
How Imaging Mass Cytometry™ is used to understand the localization of tumor antigen-specific T cells
About the presenter:
Hiroyoshi Nishikawa, MD, PhD Division Chief of Cancer Immunology
Research Institute/Exploratory Oncology Research & Clinical Trial Center (EPOC)
National Cancer Center, Tokyo/Chiba, Japan
Professor of Immunology
Nagoya University Graduate School of Medicine
Professor Hiroyoshi Nishikawa is Division Chief of Cancer Immunology at the Research Institute/Exploratory Oncology Research & Clinical Trial Center (EPOC) at the National Cancer Center Japan and a Professor of Immunology at Nagoya University Graduate School of Medicine. Throughout his career his research has focused on tumor biology, tumor therapeutics and immunology. Currently his group aims to comprehensively investigate immune cells such as CD4+, CD8+ T cells and macrophages, cancer cells and environmental factors to clarify the molecular mechanisms that control immune balances in a tumor microenvironment.
For Research Use Only. Not for use in diagnostic procedures.
Deep Immunophenotyping of Cancer Microenvironments by Imaging Mass Cytometry
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WEBINAR
Noel de Miranda, PhD discusses Deep Immunophenotyping of Cancer Microenvironments by Imaging Mass Cytometry
Multiplex immunophenotyping technologies are indispensable for a deeper understanding of biological systems. Until recently, high-dimensional cellular analyses implied the loss of tissue context because most were performed in single-cell suspensions. In this webinar Noel de Miranda introduces how the advent of Imaging Mass Cytometry™ (IMC™) has enabled highly multiplexed immunohistochemistry of frozen and formalin-fixed, paraffin-embedded (FFPE) tissues using proven CyTOF® technology.
About this webinar:
In this presentation from the July 2020 Fluidigm Oncology and Infectious Disease Virtual Summit, de Miranda discusses:
Details of how his team developed an optimized workflow for maximum antibody performance
Insights as to how IMC can be implemented in other laboratories
Methodological challenges that were overcome to develop a large antibody panel to preserve signal intensity and specificity of antigen detection
Hear de Miranda outline why the IMC panel developed here is an excellent immune monitoring tool that can be readily applied in the context of research and clinical trials.
About the presenter:
Noel de Miranda, PhD Principal Investigator
Leiden University Medical Center
Noel de Miranda is a Principal Investigator in the Department of Pathology at the Leiden University Medical Center and is the Project Leader for the Immunogenomics group. His group aims to develop innovative therapies focused on stimulating the immune system to recognize and eliminate cancer cells. Currently the research focuses on the development of neoantigen-targeted therapies for patients diagnosed with cancers with low-mutational burden and the deep immuno-phenotyping of colorectal and pancreatic cancers and discovery of immune cell subsets with anti-tumor activity.
For Research Use Only. Not for use in diagnostic procedures.
Webinar | Defining the Spatial Localization of Human Innate Cell Precursors
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WEBINAR
Hear Emily Mace, PhD, of Columbia University Irving Medical Center give her presentation titled Defining the Spatial Localization of Human Innate Cell Precursors in tissue by Imaging Mass Cytometry.
Emily Mace is working to understand the spatial relationship between natural killer (NK) cell precursors and other immune cells and relevant cytoarchitecture using Imaging Mass Cytometry™. Human NK cells are critical immune effector cells that control viral infection and malignancy. Their importance is underscored by the severe disease that occurs when their development is impaired or dysregulated. In the work she describes here, her lab seeks to understand the molecular events that drive NK cell differentiation from common lymphoid progenitors.
About this webinar:
In this webinar presented initially at the CYTO® 2020 virtual meeting, Mace introduces her lab’s approach to answer key questions about human NK cell development. She discusses:
The spectrum of developmental intermediates that show increasingly restricted lineage potential that can be isolated from tissue and give rise to mature, functional NK cells
The spatial localization of NK cell developmental intermediates within human secondary lymphoid tissue
Preliminary findings that provide an exciting foundation for the first in situ roadmap of human innate immune cell development in tissue
About the presenter:
Emily Mace, PhD Assistant Professor of Pediatric Immunology
Columbia University Irving Medical Center
Emily Mace is an Assistant Professor of Pediatrics at Columbia University Irving Medical Center in New York. She studies human natural killer cell development, particularly with quantitative image analysis and cell biological approaches. This includes the use of highly spatially and temporally resolved and super-resolution microscopy to understand interactions between NK cell precursors and the microenvironment. She also identifies novel requirements for human NK cell development through the identification and study of rare patients with NK cell deficiencies. This has included the characterization of NK cell functional and cell biological phenotypes associated with MCM10, GATA2, IRF8 and coronin 1A deficiencies.
For Research Use Only. Not for use in diagnostic procedures.
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