Sequential identification of apoptotic and necrotic cells on individual cytospin preparations

Here, we describe a simple methodology which allows the consecutive differentiation of apoptotic and necrotic cells on the same cytospin preparation of fresh and cultured cells. In this methodology, necrotic cells are initially identified on the cytospin preparations using trypan blue followed by the identification of in situ DNA fragmentation using the TUNEL assay. Identification of trypan blue and TUNEL positive cells in the same section permits the simultaneous assessment of necrotic and apoptotic cells.