Detecting Apoptosis by Flow Cytometry - Where are we after 25 years?

It has been over 25 years since the earliest flow cytometric techniques for analyzing apoptosis were first reported.  So where are we now?   Flow cytometric methods for the physical manifestations of apoptosis, including DNA damage, changes in and loss of cell membrane permeability, and organelle changes (particularly mitochondria) are now well known and widely used.  In this talk, we will concentrate on methods aimed at identifying the equally important signal transduction elements of apoptosis, especially the activation of caspases, enzymes critical for both the signaling and effecter functions of cell death.  We will examine the practical benefits and pitfalls of the many fluorogenic caspase detection reagents available to researchers for analyzing apoptosis, and how they can be combined with other apoptosis assays for form powerful multi-parametric assays for cell death.  Apoptosis research has also benefited from the growing field of image cytometry, where cytometric and morphological data can be combined to provide even more insight into this critical process.  We will also discuss developments in this expanding field.  Finally, we will discuss autophagy, a process originally thought to be a component of apoptosis but now found to be distinct.  This important process is only now becoming accessible to analysis by flow cytometry. 

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