Determining the structure-mechanics relationships of dense microtubule networks with confocal microscopy and magnetic tweezers-based microrheology.

摘要: The microtubule (MT) cytoskeleton is essential in maintaining the shape, strength, and organization of cells. Its spatiotemporal fundamental for numerous dynamic biological processes, mechanical stress within MT provides an important signaling mechanism mitosis neural development. This raises questions about relationships between structure mechanics complex structures. In vitro, reconstituted cytoskeletal networks provide a minimal model cell while also providing testing ground polymer physics stiff gels. Here, we describe our development implementation broad tool kit to study structure–mechanics networks, including protocols assembly entangled cross-linked fluorescence imaging, microstructure characterization, construction calibration magnetic tweezers devices, data collection analysis. particular, present design three neodymium iron boron (NdFeB)-based devices optimized use with networks: (1) high-force that enable application nano-Newton forces possible meso- macroscale materials characterization; (2) ring-shaped NdFeB-based oscillatory microrheology measurements; (3) portable direct visualization microscale deformation soft under applied force.