Flow cytometry is a powerful technique that allows researchers to examine multiple proteins on cell populations using fluorescently labeled antibodies. Multi-parameter fluidic-based flow cytometry has been applied to multiple fields of study ranging from basic to clinical immunology, cancer biology, neuroscience, stem cell research and drug discovery. In order to maximize the quality of results obtained using this technology, researchers should be familiar with the basics of flow cytometry when setting up an experiment. In this series of webinars, we will provide investigators with the necessary tools to design, run, and analyze a multi-parameter flow cytometric experiment.
Webinar 1: Relax, and Go with the Flow: Demystifying Multi-Parameter Flow Cytometry
In this introductory level webinar, you will learn the basics of how flow cytometry works including how to optimally set up and run a multi-parameter experiment. We will highlight such topics as:
- spectral overlap and how compensation is used to correct this issue
- how the surface density of cellular antigens influences fluorochrome choice
- basic data analysis
To illustrate these points, we will feature a short video on how to run a basic multicolor flow experiment on human natural Tregs, using techniques which can be applied to any field of study.
Webinar 2: Turning Flow Cytometry Upside Down and Inside Out: Cellular Function Revealed
In the second webinar, you will learn how to apply basic concepts of flow cytometry to topics such as cell proliferation, dead cell exclusion, and intracellular staining techniques for transcription factors and phospho-proteins. In particular, you will learn the importance of:
- titrating cell proliferation dyes and flow antibodies for optimal results
- how dead cell exclusion and staining temperatures of surface markers can influence interpretation of your data
- different fixation and permeabilization buffers and their use in detecting transcription factors and secreted cytokines vs. phospho-proteins
An in-depth discussion of these techniques will be illustrated in a multi-parameter flow experiment examining human Memory T cell subsets.
|Knockout Validated Proliferation Markers