Early Detection of Apoptosis in Living Cells by Fluorescence Correlation Spectroscopy

Michelle Martinez, Randall Reif, Dimitrios Pappas

Research output: Contribution to journalArticlepeer-review

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, while cells with cleaved probe showed diffusion and molecular brightness characteristics of rhodamine 110. Using FCS measurements, cells were identified as apoptotic based on the presence of autocorrelated fluorescence, average molecular brightness (η), and molecular dwell time (τD). Apoptotic cells identified in this manner were detected as early as 45 minutes 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 anal
Original languageEnglish
Pages (from-to)1177-1185
JournalAnalytical and Bioanalytical Chemistry
StatePublished - 2010

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