One approach to applying precise forces to cells is atomic force microscopy (AFM).
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Thus, the magnitude of confinement force required to prevent mitosis and the mechanisms of cell response to such forces are not known. Confining cell rounding below 5-8 µm with microfabricated polydimethylsiloxane (PDMS) chambers perturbs mitotic progression in several cell types ( 2, 10), but the forces required to do this remain unquantified. Despite the importance of cell rounding in mitotic progression and tissue organization, the mechanical robustness of mitotic cells remains poorly investigated even in vitro, probably due to a paucity of suitable experimental tools that can apply precise forces to poorly adherent cells. Abnormal mitotic cell shape can have adverse consequences for chromosome segregation and tissue growth ( 9), in some cases contributing to tumorigenesis ( 7). Recent studies in the epithelium and epidermis of various organisms indicate that mitotic cell rounding is involved in tissue organization, development, and homeostasis ( 4– 8). In mitosis, eukaryotic cells down-regulate focal adhesions and increase their cortical tension and intracellular pressure, thereby generating force to round up against external impediments ( 1– 3). Thus, we reveal how the mechanical properties of mitotic cells and their response to external forces are linked to mitotic progression under conditions of mechanical confinement. Our results suggest that a buildup of actomyosin-dependent cortical tension and intracellular pressure precedes mechanical failure, or herniation, of the cell cortex at the yield force. Yield forces are coincident with a nonlinear drop in cell height potentiated by persistent blebbing and loss of cortical F-actin homogeneity. We identify three force regimes according to the cell response: small forces (∼5 nN) that accelerate mitotic progression, intermediate forces where cells resist confinement (50-100 nN), and yield forces (>100 nN) where a significant decline in cell height impinges on microtubule spindle function, thereby inhibiting mitotic progression. Here we introduce ion beam-sculpted microcantilevers that enable precise force-feedback–controlled confinement of single cells while characterizing their progression through mitosis.
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I wrote about how its fit into my digital life in a blog post shortly after I first found it.Despite the importance of mitotic cell rounding in tissue development and cell proliferation, there remains a paucity of approaches to investigate the mechanical robustness of cell rounding. The Daily: Minbox is my new favorite tool for delivering files, small and large, when a client calls out for them. He uses his background in production to work with everyone from the DP to the Script Supervisor to get the best of the footage in the cut, often collaborating from the pre-production stages all the way through. The Download: Natums started in studio broadcast, worked through different roles in production until he found himself sitting comfy in post-production. The Quote: “ Client wants random file, client has random file.” #FreakingFast Facts: Video editor, JKL Jockey, addicted to Twitter, nocturnal. Just drag, drop, hit send, and forget about it. But you won’t have to wait for your uploads to finish.
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