title: Single cell viability measurements in 3D scaffolds using in situ label free imaging by optical coherence microscopy.
abstract: The focus on creating tissue engineered constructs of clinically relevant sizes requires new approaches for monitoring construct health during tissue development. A few key requirements are that the technology be in situ, non-invasive, and provide temporal and spatial information. In this work, we demonstrate that optical coherence microscopy (OCM) can be used to assess cell viability without the addition of exogenous probes in three-dimensional (3D) tissue scaffolds maintained under standard culture conditions. This is done by collecting time-lapse images of speckle generated by sub-cellular features. Image cross-correlation is used to calculate the number of features the final image has in common with the initial image. If the cells are live, the number of common features is low. The number of common features approaches 100% if the cells are dead. In control experiments, cell viability is verified by the addition of a two-photon fluorescence channel to the OCM. Green fluorescent protein transfected human bone marrow stromal cells cultured in a transparent poly(ethylene glycol) tetramethacrylate hydrogel scaffold is used as the control system. Then, the utility of this approach is demonstrated by determining L929 fibroblast cell viability in a more challenging matrix, collagen, an optical scatterer. These results demonstrate a new technique for in situ mapping of single cell viability without any exogenous probes that is capable of providing continuous monitoring of construct health.
authors.affiliation: Polymers Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA. joy.dunkers@nist.gov
authors.forename: Joy P
authors.initials: JP
authors.lastname: Dunkers
authors.forename: Young Jong
authors.initials: YJ
authors.lastname: Lee
authors.forename: Kaushik
authors.initials: K
authors.lastname: Chatterjee
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D016000:Cluster Analysis; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology; D054457:Tissue Scaffolds; D055815:Young Adult
chemical_list: D020100:Hydrogels