Temporal dynamics of receptor-induced apoptosis in an affinity microdevice

Dimitrios Pappas, Randall D Reif, Charmaine Aguas, Michelle M Martinez

Research output: Contribution to journalArticle

Abstract

The temporal dynamics of Fas-induced apoptosis is elucidated. Jurkat cells are captured on the affinity surface of a microdevice coated with anti-CD95, an antibody known to induce apoptosis in cells via the extrinsic (caspase 8) pathway. The timing of apoptosis induction is controlled by the binding of the cells to the surface. Once bound, the cells are continuously stained with the caspase probe, L-bisaspartic acid rhodamine 110 (D2R), and the fluorescence of the cells was monitored for 6 h by light microscopy. This approach normalizes the temporal dynamics for each cell, as the binding event is also the start of apoptosis. In addition to providing the number of apoptotic cells over time, the fluorescence of individual cells can be monitored, providing information about the timing of caspase activity in each cell. The rate of caspase cleavage of D2R in each cell is also measured and shows good agreement between the cells in a given population. The effects of the caspase inhibitor, z-
Original languageEnglish
Pages (from-to)3387–3396
JournalAnalytical and Bioanalytical Chemistry
StatePublished - 2010

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