18 February 2011 Chronic label-free volumetric photoacoustic microscopy of melanoma cells in scaffolds in vitro
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Visualizing cells in three-dimensional (3D) scaffolds has been one of the major challenges in tissue engineering. Current imaging modalities have limitations. Microscopy, including confocal microscopy, cannot penetrate deeply (> 300 μm) into the scaffolds; X-ray micro-computed tomography (micro-CT) requires staining of the structure with a toxic agent such as osmium tetroxide. Here, we demonstrate photoacoustic microscopy (PAM) of the spatial distribution and temporal proliferation of melanoma cells inside three-dimensionally porous scaffolds with thicknesses over 1 mm. Melanoma cells have a strong intrinsic contrast which is easily imaged by label-free PAM with high sensitivity. Spatial distributions of the cells in the scaffold were well-resolved in PAM images. Moreover, we chronically imaged the same cell/scaffold constructs at different time points over 2 weeks. The number of cells in the scaffold was quantitatively measured from the PAM volumetric information. The cell proliferation profile obtained from PAM correlated well with that obtained using the traditional 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. We believe that PAM will become a useful imaging modality for tissue engineering applications, especially when thick scaffold constructs are involved, and that this modality can also be extended to image other cell types labeled with contrast agents.
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Xin Cai, Xin Cai, Yu Zhang, Yu Zhang, Chulhong Kim, Chulhong Kim, Sung-Wook Choi, Sung-Wook Choi, Younan Xia, Younan Xia, Lihong V. Wang, Lihong V. Wang, "Chronic label-free volumetric photoacoustic microscopy of melanoma cells in scaffolds in vitro", Proc. SPIE 7899, Photons Plus Ultrasound: Imaging and Sensing 2011, 789926 (18 February 2011); doi: 10.1117/12.873226; https://doi.org/10.1117/12.873226

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