Abstract:
Cell death is important in physiology, and can happen as a result of structural damage, or as
a sequence of programmed cellular processes known as apoptosis. Pathogenic alterations
in apoptosis occur in a number of diseases, including cancer, viral infections, autoimmune
diseases, immunodeficiencies, and degenerative conditions. Developing accurate and
reproducible laboratory methods for inducing and detecting apoptosis is vital for research
into these conditions. A number of methods are employed to detect cell death, including
DNA fragmentation, the TUNEL assay, and electron microscopy although each has its limitations. Flow cytometry allows for the distinction between live, early apoptotic, late apoptotic
and necrotic cells. In this protocol we successfully induce apoptosis using chemical treatment and treatment with low pH in solid tumour cell lines, and have optimized detection
using the Annexin V/PI apoptosis assay.
