Apoptosis and Necrosis using Flow Cytometry
Using Flow Cytometry an investigator can study a cell
population and see what percentage are undergoing apoptosis or
already died. A cell population is treated for a set amount of
and then fixed. Once fixed the population is stained using
Iodide (PI) which stains dead cells and a FITC labeled
V) to phosphatidylserine (PS). PS is a phospholipid normally
the cell membrane, but which is transferred to the outer
the cell membrane and is used as an indicator of apoptosis.
flow data looks like this:
From this experiment we can see that 29.8% of
is apoptotic, with 2.1% of the population being already dead
a compromised membrane.
Another experiment which can be performed is
assay. This assay measures DNA fragmentation using an enzyme
biotinylated uridine. A FITC labeled antibody against biotin
the uridine. When run through the flow cytometer the FITC
You can use this assay to see how many of the cells have
DNA, another indication of apoptosis.