Description
- Abstract:
- Data from: Into the depths: Techniques for in vitro three-dimensional microtissue visualization Pranita K. Kabadi, Marguerite M. Vantangoli, April L. Rodd, Elizabeth Leary, Samantha J. Madnick, Jeffrey R. Morgan, Agnes Kane, and Kim Boekelheide BioTechniques 2015 59:5, 279-286 doi: 10.2144/000114353. Here we optimized histology and microscopy techniques to overcome the constraints of 3-D imaging. For morphological assessment of 3-D microtissues of several cell types, different time points, and different sizes, a two-step glycol methacrylate embedding protocol for evaluating 3-D microtissues produced using agarose hydrogels improved resolution of nuclear and cellular histopathology characteristic of cell death and proliferation. Additional immunohistochemistry, immunofluorescence, and in situ immunostaining techniques were successfully adapted to these microtissues and enhanced by optical clearing. Utilizing the ClearT2 protocol greatly increased fluorescence signal intensity, imaging depth, and clarity, allowing for more complete confocal fluorescence microscopy imaging of these 3-D microtissues compared with uncleared samples. The refined techniques presented here address the key challenges associated with 3-D imaging, providing new and alternative methods in evaluating disease pathogenesis, delineating toxicity pathways, and enhancing the versatility of 3-D in vitro testing systems in pharmacological and toxicological applications.
- Notes:
- This research is supported by NIEHS Training Grant T32 ES007272, the NIEHS Superfund Research Program P42 ES013660, BP/The Gulf of Mexico Research Initiative through the Consortium for Molecular Engineering of Dispersant Systems (CMEDS), and the generous gift of Donna McGraw Weiss ‘89 and Jason Weiss.
Access Conditions
- Use and Reproduction
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- This work is licensed under a Creative Commons Attribution 4.0 International License
Citation
Kabadi, Pranita K., Vantangoli, Marguerite M., Rodd, April L., et al.,
"Image and Video Data Files from 'Into the Depths: Techniques for in vitro Three-Dimensional Microtissue Visualization'"
(2015).
Brown Superfund Data Products, Kane Lab Data Archive, Brown University Open Data Collection.
Brown Digital Repository. Brown University Library.
https://doi.org/10.26300/h39v-zg24
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Brown Superfund Data Products
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Kane Lab Data Archive
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Brown University Open Data Collection
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