Department of Physics, University of California San Diego, La Jolla, California, United States of America.
PLoS One. 2011;6(9):e25534. doi: 10.1371/journal.pone.0025534. Epub 2011 Sep 30.
Thin layers of gels with mechanical properties mimicking animal tissues are widely used to study the rigidity sensing of adherent animal cells and to measure forces applied by cells to their substrate with traction force microscopy. The gels are usually based on polyacrylamide and their elastic modulus is measured with an atomic force microscope (AFM). Here we present a simple microfluidic device that generates high shear stresses in a laminar flow above a gel-coated substrate and apply the device to gels with elastic moduli in a range from 0.4 to 300 kPa that are all prepared by mixing two components of a transparent commercial silicone Sylgard 184. The elastic modulus is measured by tracking beads on the gel surface under a wide-field fluorescence microscope without any other specialized equipment. The measurements have small and simple to estimate errors and their results are confirmed by conventional tensile tests. A master curve is obtained relating the mixing ratios of the two components of Sylgard 184 with the resulting elastic moduli of the gels. The rigidity of the silicone gels is less susceptible to effects from drying, swelling, and aging than polyacrylamide gels and can be easily coated with fluorescent tracer particles and with molecules promoting cellular adhesion. This work can lead to broader use of silicone gels in the cell biology laboratory and to improved repeatability and accuracy of cell traction force microscopy and rigidity sensing experiments.
具有模拟动物组织力学性能的薄层凝胶广泛用于研究贴壁动物细胞的刚性感知,并通过牵引力显微镜测量细胞对其基质施加的力。这些凝胶通常基于聚丙烯酰胺,其弹性模量是用原子力显微镜(AFM)测量的。在这里,我们提出了一种简单的微流控装置,该装置可在凝胶覆盖基底上方的层流中产生高剪切应力,并将该装置应用于弹性模量在 0.4 至 300 kPa 范围内的凝胶,这些凝胶均通过混合两种透明商业硅酮 Sylgard 184 的成分制备而成。通过在宽场荧光显微镜下跟踪凝胶表面上的小球,无需任何其他专用设备即可测量弹性模量。测量具有较小且易于估计的误差,并且其结果通过常规拉伸测试得到确认。获得了与 Sylgard 184 的两种成分的混合比与凝胶的最终弹性模量相关的主曲线。硅酮凝胶的刚性比聚丙烯酰胺凝胶不易受到干燥、溶胀和老化的影响,并且可以很容易地用荧光示踪颗粒和促进细胞黏附的分子进行涂层。这项工作可以促进硅酮凝胶在细胞生物学实验室中的更广泛应用,并提高细胞牵引力显微镜和刚性感知实验的重复性和准确性。