CONTENTS

Introduction..................................................................................................................................... 35

Morphological Assays of Apoptosis...............................................................................................

Electron Microscopy................................................................................................................ 37

Light Microscopy..................................................................................................................... 38

Fluorescence Microscopy after DNA Staining........................................................................ 38

Detection of DNA Fragmentation................................................................................................... 38

Conventional Agarose Gel Electrophoresis............................................................................. 39

Extraction of Small Fragments................................................................................................ 39

Flow Cytometry........................................................................................................................ 40

Filtration Assays...................................................................................................................... 40

Terminal Deoxyribonucleotidyl Transferase-Mediated dUTP

Nick-End Labeling (TUNEL)................................................................................................... 41

Comet Assay............................................................................................................................. 41

Caspase Activation.......................................................................................................................... 42

Activity Assays........................................................................................................................

Cleavage of Procaspases and Substrates.................................................................................. 44

Affinity Labeling...................................................................................................................... 45

Immunodetection of Activated Caspases................................................................................. 46

Annexin V Staining......................................................................................................................... 46

Flow Cytometry-Based Assays....................................................................................................... 47

Altered Cell Size...................................................................................................................... 47

Loss of Mitochondrial Transmembrane Potential (∆Ψ_m)....................................................... 48

Dye Uptake............................................................................................................................... 48

Acknowledgments........................................................................................................................... 49

References....................................................................................................................................... 49