Early detection of apoptosis in living cells by fluorescence correlation spectroscopy

Michelle M. Martinez, Randall D. Reif, Dimitri Pappas

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

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 (η), and molecular dwell time (τ 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.

Original languageEnglish
Pages (from-to)1177-1185
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Volume396
Issue number3
DOIs
StatePublished - 2010

Keywords

  • Apoptosis
  • Cancer
  • Caspases
  • Fluorescence correlation spectroscopy
  • Single-cell analysis

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