Campbell Benjamin E, Mok Stephanie, Moraes Christopher
Department of Chemical Engineering, McGill University, Montréal, QC, Canada.
Department of Biomedical Engineering, McGill University, Montréal, QC, Canada.
Methods Mol Biol. 2023;2614:237-246. doi: 10.1007/978-1-0716-2914-7_14.
Local tissue scale mechanical properties are essential for understanding cell fate and function; however, few methods to measure stiffness at this length scale exist, and applications in 3D tissues can present further challenges. To address this need, microgel-based sensors fabricated out of the thermally responsive hydrogel poly(N-isopropylacrylamide) were developed allowing internal architectures of tissues to be mapped by optically measuring microgel response when actuated in a matrix. These robust probes are widely applicable for in vitro and in vivo studies of tissue mechanics providing tissues can be fluorescently imaged. Here we describe the fabrication of these thermally responsive hydrogel sensors, calibration of the microgels using phantom tissues, and image processing techniques used to make the measurements.
局部组织尺度的力学特性对于理解细胞命运和功能至关重要；然而，在这个长度尺度上测量硬度的方法很少，并且在三维组织中的应用可能会带来更多挑战。为了满足这一需求，开发了由热响应水凝胶聚（N-异丙基丙烯酰胺）制成的基于微凝胶的传感器，当在基质中被激活时，通过光学测量微凝胶的响应来绘制组织的内部结构。这些坚固的探针广泛适用于组织力学的体外和体内研究，前提是组织可以进行荧光成像。在这里，我们描述了这些热响应水凝胶传感器的制造、使用虚拟组织对微凝胶进行校准以及用于进行测量的图像处理技术。
