FACS

In the Fluorescence-automated Cell Sorting (FACS) Simulation, you will learn the basics of flow cytometry and how to use a flow cytometer with fluorescence detection. Your goal is to sort a pool of Escherichia coli expressing Green Fluorescent Protein (GFP) at different levels after it has been previously engineered. You will set up the equipment, learn how the cytometer sorts the cells, interpret your results, and clean and shut down the equipment. Will you be able to perform all the steps correctly and sort the cells depending on their GFP expression level?

Following is a list of the relevant theory pages:

• FACS definition

• Fluidics system

  • Flow buffer

  • Closed-loop nozzle

  • Droplet break-off point

• Optics system

  • Interrogation point

  • FACS signal measurements

• Electronics system

  • Electrostatic sorting

  • Drop delay

• Data analysis

  • Gating

• FACS general protocol

• Fluorescence

  • GFP protein

• Beta-carotene