AI Article Synopsis
- Serial block face scanning electron microscopy (SBF-SEM) enables detailed 3D analysis of cells, particularly useful for studying microtubules in the mitotic spindle of human cells.
- The study highlights a new workflow that combines light microscopy with SBF-SEM to achieve improved visualization of microtubules, bridging fibers, and cellular membranes during mitosis.
- The research also introduces computational tools that analyze the spatial arrangement of microtubules and shows their sensitivity to TACC3 levels affecting microtubule order in mitotic spindles.
Article Abstract
Serial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light-SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present four examples of uses for this workflow that are not practical by light microscopy and/or transmission electron microscopy. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus; and fourth, volumetric analysis of kinetochores. Our workflow also includes new computational tools for exploring the spatial arrangement of microtubules within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6173286 | PMC |
| http://dx.doi.org/10.1242/jcs.203877 | DOI Listing |
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