title: Fluorescent oxygen sensitive microbead incorporation for measuring oxygen tension in cell aggregates.
abstract: Molecular oxygen is a main regulator of various cell functions. Imaging methods designed as screening tools for fast, in situ, 3D and non-interfering measurement of oxygen tension in the cellular microenvironment would serve great purpose in identifying and monitoring this vital and pivotal signalling molecule. We describe the use of dual luminophore oxygen sensitive microbeads to measure absolute oxygen concentrations in cellular aggregates. Stable microbead integration, a prerequisite for their practical application, was ensured by a site-specific delivery method that is based on the interactions between streptavidin and biotin. The spatial stability introduced by this method allowed for long term measurements of oxygen tension without interfering with the cell aggregation process. By making multiple calibration experiments we further demonstrated the potential of these sensors to measure local oxygen tension in optically dense cellular environments.
authors.affiliation: Department of Metallurgy and Materials Engineering, KU Leuven, Kasteelpark Arenberg 44, Box 2450, 3001 Leuven, Belgium.
authors.forename: Dennis
authors.initials: D
authors.lastname: Lambrechts
authors.forename: Maarten
authors.initials: M
authors.lastname: Roeffaers
authors.forename: Greet
authors.initials: G
authors.lastname: Kerckhofs
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Roberts
authors.forename: Johan
authors.initials: J
authors.lastname: Hofkens
authors.forename: Tom
authors.initials: T
authors.lastname: Van de Putte
authors.forename: Hans
authors.initials: H
authors.lastname: Van Oosterwyck
authors.forename: Jan
authors.initials: J
authors.lastname: Schrooten
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002449:Cell Aggregation / Q000502:physiology; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008863:Microspheres; D057054:Molecular Imaging / Q000379:methods*; D010092:Oximetry / Q000379:methods*; D010100:Oxygen / Q000032:analysis*; D010101:Oxygen Consumption / Q000502:physiology*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D010100:Oxygen