Cell Death Analysis in Retinal Cultures

Abstract

Evaluating cell death is essential when investigating neurodegeneration and neuroprotection in the retina. Cell death assays provide us with a means to identify and quantify dying cells in a population. Terminal dUTP nick end-labeling (TUNEL) is one method used for the identification of dying cells. This technique is based upon the enzymatic incorporation of fluorescently tagged DNA base pairs to fragmented DNA. In this chapter, we describe two different techniques employing TUNEL. The first method uses TUNEL to analyze cell death in cultured retinal explants by fluorescence microscopy. The second technique describes a method for measuring cell death in a retinal cell line by flow cytometry.