Intrinsic fluorescence and redox changes associated with apoptosis of primary human epithelial cells

Jonathan M. Levitt, Amy Baldwin, Antonios Papadakis, Sameer Puri, Joanna Xylas, Karl Münger, Irene Georgakoudi

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Apoptosis plays a key role in the development and maintenance of human tissues. This process has been studied traditionally in cells that are stained with exogenous fluorophores. These approaches affect cell viability, and thus are ill-suited for in vivo applications. We present an imaging approach that can identify apoptotic cells in living cell populations based on detection and quantification of distinct changes in the intensity and localization of cellular autofluorescence. Specifically, we acquire NAD(P)H, FAD, and redox ratio autofluorescence images of primary keratinocytes following 1,9, 14, and 18 h of treatment with cisplatin, a known apoptosis-inducing chemotherapy agent. We find that intense autofluorescence combined with a low redox fluorescence ratio is progressively confined to a gradually smaller perinuclear cytoplasmic region with cisplatin treatment. Studies with exogenous nuclear fluorophores demonstrate that these autofluorescence changes occur at early stages of apoptosis. Additional costaining experiments suggest that this strongly autofluorescent, highly metabolically active perinuclear ring represents a subpopulation of mitochondria that are mobilized in response to the apoptotic stimulus and may provide the energy required to execute the final apoptotic steps. Thus, autofluorescence localization changes could serve as a sensitive, noninvasive indicator of early apoptosis in vivo.

Original languageEnglish (US)
Article number064012
JournalJournal of Biomedical Optics
Volume11
Issue number6
DOIs
StatePublished - Nov 1 2006
Externally publishedYes

Fingerprint

apoptosis
Cell death
Fluorescence
Apoptosis
fluorescence
Fluorophores
Cells
Cisplatin
Flavin-Adenine Dinucleotide
Mitochondria
mitochondria
Chemotherapy
chemotherapy
viability
NAD
stimuli
maintenance
Tissue
Imaging techniques
Oxidation-Reduction

Keywords

  • Apoptosis
  • Fluorescence
  • Microscopy
  • Optical diagnosis
  • Redox ratio

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Biomedical Engineering

Cite this

Intrinsic fluorescence and redox changes associated with apoptosis of primary human epithelial cells. / Levitt, Jonathan M.; Baldwin, Amy; Papadakis, Antonios; Puri, Sameer; Xylas, Joanna; Münger, Karl; Georgakoudi, Irene.

In: Journal of Biomedical Optics, Vol. 11, No. 6, 064012, 01.11.2006.

Research output: Contribution to journalArticle

Levitt, Jonathan M. ; Baldwin, Amy ; Papadakis, Antonios ; Puri, Sameer ; Xylas, Joanna ; Münger, Karl ; Georgakoudi, Irene. / Intrinsic fluorescence and redox changes associated with apoptosis of primary human epithelial cells. In: Journal of Biomedical Optics. 2006 ; Vol. 11, No. 6.
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