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Issue 17, 2013
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High temporal resolution fluorescence measurements of a mitochondrial dye for detection of early stage apoptosis

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Abstract

In the present study, early stage apoptosis is explored with high temporal resolution. In addition to monitoring early apoptosis induction in single cells by ultrasensitive confocal fluorescence microscopy (UCFM), the mitochondrial protein release kinetics was explored. The current study shows development and optimization of a novel, rapid apoptosis assay to explore the earliest changes in cells by the intrinsic apoptosis pathway. We show that early apoptotic changes in the mitochondria begin nearly simultaneously with the addition of an apoptosis-inducing drug, such as staurosporine. With a temporal resolution of five minutes, this non-invasive analytical technique can elucidate the earliest apoptotic events in living cells. Moreover, our results show that the mitochondrial inter-membrane proteins are not involved in the extrinsic pathway of Ramos cells mediated by an anti-CD95 antibody. Additional techniques such as light microscopy and flow cytometry were employed to confirm the results obtained by ultrasensitive confocal fluorescence microscopy. The results of this study help to understand the earliest mechanisms of apoptosis induction in cells, enabling new methods of drug testing and dose–response analyses.

Graphical abstract: High temporal resolution fluorescence measurements of a mitochondrial dye for detection of early stage apoptosis

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Publication details

The article was received on 07 Jun 2013, accepted on 17 Jun 2013 and first published on 18 Jun 2013


Article type: Paper
DOI: 10.1039/C3AN01142A
Analyst, 2013,138, 4892-4897

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    High temporal resolution fluorescence measurements of a mitochondrial dye for detection of early stage apoptosis

    D. Iyer, R. D. Ray and D. Pappas, Analyst, 2013, 138, 4892
    DOI: 10.1039/C3AN01142A

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