title: Assesment of apoptosis induced changes in scattering using optical coherence tomography.
abstract: The aim of this study is to identify changes in scattering with optical coherence tomography (OCT) and relate these measurements with mitochondrial changes during the initiation of apoptosis. Human retinal pigment epithelial cells were cultured and apoptosis was induced using 10% alcohol. Using the attenuation coefficient and backscattering, changes were measured during cell death in a cell-pellet and monolayer respectively. To confirm apoptosis, fluorescent activated cell sorting was used. Mitochondrial activity during apoptosis was assessed using an oxidative stress assay and fluorescent confocal microscopy. Pelleted apoptotic cells measured with OCT showed a clear rise while untreated cells showed a very small increase in attenuation coefficient. Monolayered apoptotic cells displayed a distinct increase, while untreated cells showed a small increase in the backscattering. Apoptosis was confirmed by FACS experiments. Mitochondrial changes during the onset of apoptosis were also measured. The results demonstrate that apoptotic cell death could be monitored in real-time by OCT. Changes in the scattering after induction of apoptosis are likely to be related to changes in the intracellular morphology. Oxidative stress-induced mitochondrial swelling could be responsible for the initial increase, while cell blebbing and secondary necrosis subsequently for the observed decrease in scattering.
authors.affiliation: Biomedical Engineering and Physics, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands. d.m.debruin@amc.nl.|Urology, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands. d.m.debruin@amc.nl.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: de Bruin
authors.affiliation: Experimental Surgery, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands.
authors.forename: Mans
authors.initials: M
authors.lastname: Broekgaarden
authors.affiliation: Biomedical Engineering and Physics, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands.
authors.forename: Martin J C
authors.initials: MJ
authors.lastname: van Gemert
authors.affiliation: Experimental Surgery, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands.
authors.forename: Michal
authors.initials: M
authors.lastname: Heger
authors.affiliation: Urology, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands.
authors.forename: Jean J
authors.initials: JJ
authors.lastname: de la Rosette
authors.affiliation: Biomedical Engineering and Physics, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands.
authors.forename: Ton G
authors.initials: TG
authors.lastname: Van Leeuwen
authors.affiliation: Biomedical Engineering and Physics, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, Netherlands.
authors.forename: Dirk J
authors.initials: DJ
authors.lastname: Faber
mesh_terms: D017209:Apoptosis; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000473:pathology*; D009336:Necrosis; D018384:Oxidative Stress; D055213:Retinal Pigment Epithelium / Q000166:cytology; D041623:Tomography, Optical Coherence
keywords: Apoptosis; Cell Death; Con-focal microscopy; FACS; Fluorescence; Mitochondria; Optical Coherence Tomography; Scattering