Abstract
BackgroundThe purpose of the study was to develop and illustrate three-dimensional (3-D) reconstruction of nuclei and intracellular lipid peroxidation in cells exposed to oxidative stress induced by quantum dots. Programmed cell death is characterized by multiple biochemical and morphological changes in different organelles, including nuclei, mitochondria and lysosomes. It is the dynamics of the spatio-temporal changes in the signalling and morphological adaptations which will ultimately determine the 'shape' and fate of the cell.ResultsWe present new approaches to the 3-D reconstruction of organelle morphology and biochemical changes in confocal live-cell images. We demonstrate the 3-D shapes of nuclei, the 3-D intracellular distributions of QDs and the accompanying lipid-membrane peroxidation, and provide methods for quantification.ConclusionThis study provides an approach to 3-D organelle and nanoparticle visualization in the context of cell death; however, this approach is also applicable more generally to investigating changes in organelle morphology in response to therapeutic interventions, stressful stimuli and internalized nanoparticles. Moreover, the approach provides quantitative data for such changes, which will help us to better integrate compartmentalization of subcellular events and to link morphological and biochemical changes with physiological outcomes.
Highlights
The purpose of the study was to develop and illustrate three-dimensional (3-D) reconstruction of nuclei and intracellular lipid peroxidation in cells exposed to oxidative stress induced by quantum dots
Our previous studies showed that Quantum dots (QDs) with unprotected surfaces are relatively unstable and can cause cell death [17]
When in serum-free medium, these cells are susceptible to QD treatment that results in significant lipid peroxidation and cell death after 24 hours
Summary
The purpose of the study was to develop and illustrate three-dimensional (3-D) reconstruction of nuclei and intracellular lipid peroxidation in cells exposed to oxidative stress induced by quantum dots. Programmed cell death is characterized by multiple biochemical and morphological changes in different organelles, including nuclei, mitochondria and lysosomes. It is the dynamics of the spatio-temporal changes in the signalling and morphological adaptations which will determine the 'shape' and fate of the cell. Quantum dots (QDs) are increasingly being used as a complement to molecular dyes in bio-imaging applications. Compared with these dyes, QDs have two special properties: size-dependent luminescence, and broad excitation but relatively narrow emission spectra [1,2,3]. Several groups have succeeded in conjugating or capping various biologically interesting ligands with QDs and have demonstrated their usefulness for different biological applications [4,5,6,7,8,9,10,11,12,13,14].
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