Ounkomol Chawin, Xie Hongtao, Dayton Paul A, Heinrich Volkmar
Department of Biomedical Engineering, University of California, Davis, Davis, California 95616, USA.
Biophys J. 2009 Feb;96(3):1218-31. doi: 10.1016/j.bpj.2008.10.047.
We present a multipurpose nanomechanical force probe that combines a sideways-mounted elastic cantilever and an optical-lever detection module with automated micropipette manipulation. It allows us to apply and measure compression, stretching, adhesion, and dissociation forces in the horizontal direction while providing a "side view" of ongoing experiments. The integrated micropipette setup facilitates the easy manipulation and mechanical interrogation of individual cells, functionalized particles, and synthetic membrane capsules. Pipette-held test objects are translated perpendicularly to and from the stationary cantilever, eliminating the need to attach them to a carrier surface and substantially reducing unwanted hydrodynamic coupling effects. Moreover, the test objects can be brought into contact with the cantilever anywhere along its length, which considerably enlarges the range of forces that can be applied with a single cantilever. Advantages of this instrument are demonstrated in example measurements of single-cell compression, membrane-tether extrusion, oligonucleotide stretching, and extraction of individual lipids from surfactant-monolayer surfaces of microbubbles.
我们展示了一种多功能纳米机械力探针,它将侧向安装的弹性悬臂和带有自动微量移液器操作的光杠杆检测模块相结合。它使我们能够在水平方向上施加和测量压缩力、拉伸力、粘附力和解离力,同时提供正在进行的实验的“侧视图”。集成的微量移液器设置便于对单个细胞、功能化颗粒和合成膜胶囊进行轻松操作和机械询问。由移液器固定的测试对象垂直于固定悬臂移动并远离固定悬臂,无需将它们附着在载体表面,并大大减少了不必要的流体动力耦合效应。此外,测试对象可以在悬臂长度的任何位置与悬臂接触,这大大扩大了使用单个悬臂可以施加的力的范围。该仪器的优势在单细胞压缩、膜系链挤出、寡核苷酸拉伸以及从微泡的表面活性剂单层表面提取单个脂质的示例测量中得到了证明。