Modular approach for resolving and mapping complex neural and other cellular structures and their associated deformation fields in three dimensions
Autor: | Jonathan B. Estrada, Rodolfo Amezcua, Eyal Bar-Kochba, Harry C. Cramer, Christian Franck, Mark T. Scimone |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Computer science Cell Culture Techniques Image processing Context (language use) Mechanotransduction Cellular General Biochemistry Genetics and Molecular Biology Rats Sprague-Dawley 03 medical and health sciences Imaging Three-Dimensional 0302 clinical medicine Software Image Processing Computer-Assisted Animals Segmentation Protocol (object-oriented programming) Cells Cultured Neurons Microscopy Confocal Artificial neural network business.industry Brain Equipment Design Modular design Biomechanical Phenomena 030104 developmental biology Transformation (function) Nerve Net business Biological system 030217 neurology & neurosurgery |
Zdroj: | Nature Protocols. 13:3042-3064 |
ISSN: | 1750-2799 1754-2189 |
Popis: | Understanding the biological implications of cellular mechanotransduction, especially in the context of pathogenesis, requires the accurate resolution of material deformation and strain fields surrounding the cells. This is particularly challenging for cells displaying branched, 3D architectures. Here, we provide a modular approach for 3D image segmentation and strain mapping of topologically complex structures. We describe how to use our approach, using neural cells and networks as an example. In addition to describing how to implement the computational analysis, we provide details of a cell culture protocol that can be used to generate neural networks for analysis and experimentation. This protocol allows for transformation of matrix-induced strains, and their full resolution across single cells or networks in three dimensions. The protocol also provides analyses to compute both the locally varying cytoskeletal strains and the average strain experienced by cells. An additional module allows spatial correlation of these strain maps with cytoskeletal features, including neurite disruptions such as neuronal blebs. Image processing and strain mapping take ≥3 h, with the exact time required being dependent on use case, software familiarity, and file size. |
Databáze: | OpenAIRE |
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