FCS Express provides a number of tools that allow you to create and graph your own data sets. Your data sets (called custom datasets) can contain data from your gates. When your gates change, so will your custom datasets and any plot derived from them. This allows you to easily create summary plots that show your entire experimental results.

 

In this tutorial, we will use custom datasets and plots to aid in the visualization of data from antibody titration experiments, to demonstrate selecting the ideal antibody concentration, so that the amount of antibody used for staining cells is the concentration that yields the best separation between positive and negative cells.

 

We will examine data from a titration experiment where seven different amounts of CD64 were used to stain peripheral blood mononuclear cells. Because CD64 is constitutively found on only macrophages and monocytes, we will examine the ratio of the intensity of fluorescence staining on the monocyte versus lymphocyte population. The negative population will be defined by the lymphocytes and the positive population by the monocytes.

 

In the course of this example, we will:

 

 

 

In the first exercise, we will define custom tokens.