Experimental Considerations with Data Sets as Examples

Now that you have a basic understanding of the principles of flow cytometry, we will delve into how powerful this technique is for detecting multiparameter surface marker expression on cells. Accurate and clean looking data is always the goal, and we’ll show how to achieve this using dead cell exclusion and Fc block. We’ll demonstrate the importance of titrating your antibodies and using the correct cell number when staining your cells. We’ll discuss how to optimize and detect cell proliferation using various reagents available on the market. We’ll illustrate the importance of temperature when detecting chemokine receptor expression and how to detect LAMP proteins using a killing assay. We’ll end with topics focusing on cellular function such as cell activation, detecting phosphorylated proteins, transcription factor expression, and secreted cytokine upregulation using Intracellular staining.