EMD Millipore eBook - 7

Analyzing Cell Death * So Many Ways to Die

rather than merely measuring it.
For information-rich cell health assessment, flow cytometry data
present obvious advantages over bulk assays such as Western blot
that rely solely on immunodetection to obtain a single readout from an
entire population. Once considered a complex instrument platform
requiring considerable scientific expertise, simplified flow cytometry
became accessible to the wider scientific community when EMD Millipore
launched the Muse® Cell Analyzer in 2012. Based on the principles
Figure 1. Apoptotic cell death may proceed via the intrinsic pathway,
or secondary to extrinsic triggers. At the onset, phosphatidylserine
(PS) translocates to the cell surface, where it may be detected
using fluorescently-labeled Annexin V. The loss of mitochondrial
membrane potential (ΔΨm ) can also herald impending apoptotic
events, and can be assayed using tools such as the Mitopotential
reagent, which fluoresces in response to ΔΨm . Both the intrinsic and
extrinsic pathways may trigger the activation of caspases, cysteine
proteases that are the principal mediators of apoptosis. In mammals,
caspases associated with apoptosis may be grouped by function
into "initiators" such as caspase-8 and -9, which cleave procaspases
to their active forms, and effector or "executioner" caspases (-3, -6,
-7) that cleave other cellular substrates to carry out the process of
programmed cell death.

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