Experiment DesignTraditional Flow Cytometry
In order to make appropriate conclusions from flow cytometry data it’s important to carefully design the experiment with proper controls. These include compensation controls, fluorescence minus one (FMO) controls, isotype controls, isoclonic controls, and unstained controls. Each control serves a unique purpose and not all controls are required for every experiment. The video below goes through the differences between these controls and how they can be utilized. If you wish to download the slides for the video, you can find them here. Below the video you’ll find a link to a blog post from a series called “Avoid Bad Data”.
For further information on compensation, see the compensation resource.
How to Identify Problems with Flow Cytometry Experiment Design – Bad Data Part 3
Two weeks ago I covered my red flags for identifying bad panel design (read that here if you missed it!). This week I’ll go over my red flags for experiment design, which mostly means controls. Controls are needed to be able to appropriately interpret experiment…
How Many Cells Should I Stain: The Impact of Cell Concentration and Antibody Concentration
When I was a graduate student, my PI asked me how many μL of a particular antibody I used per million cells, and I told him my staining conditions were “per 100 μL” not “per million cells”. This triggered a heated debate about antibody amount – he argued that the…