Early detection of apoptosis in living cells by fluorescence correlation spectroscopy

Analytical and Bioanalytical Chemistry
Michelle M MartinezDimitri Pappas

Abstract

Early detection of apoptotic cells via caspase activity is demonstrated with fast response time. Fluorescence correlation spectroscopy (FCS) is used to identify the presence of a cleaved fluorogenic probe based on the fluorescence of rhodamine 110 in Jurkat cells. FCS curves are shown to be markedly different for autofluorescent (non-apoptotic) cells, whereas cells with cleaved probe showed diffusion and molecular brightness characteristic of rhodamine 110. Using FCS measurements, cells were identified as apoptotic on the basis of the presence of autocorrelated fluorescence, average molecular brightness (eta), and molecular dwell time (tau (D)). Apoptotic cells identified in this manner were detected as early as 45 min after induction. Unlike other methods with similar identification times, such as western blotting and electron microscopy, cells remain viable for further analysis. This multi-parameter approach is rapid, flexible, and does not require transfection of the cells prior to analysis, enabling apoptosis to be identified early in a wide variety of cell types.

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Citations

Jan 7, 2011·Analytical Chemistry·Peng LiDimitri Pappas
Jul 17, 2012·Cytotechnology·Alexander P Demchenko
Jan 16, 2018·Journal of Occupational and Environmental Medicine·Jonathan E Thompson
Oct 27, 2017·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Sabrina MariottiRoberto Nisini
Mar 3, 2010·Analytical and Bioanalytical Chemistry·Randall D ReifDimitri Pappas
Feb 6, 2014·The Analyst·Kerstin GallerUte Neugebauer
Aug 9, 2020·European Journal of Pharmacology·Ping XuXijie Guo

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis