Lehigh University, Department of Chemistry, 6 E. Packer Ave, Bethlehem, PA 18015, USA.
Lab Chip. 2011 Dec 21;11(24):4248-59. doi: 10.1039/c1lc20627c. Epub 2011 Nov 3.
We demonstrated the application of a simple electrode geometry for dielectrophoresis (DEP) on colloidal probes as a form of molecular force spectroscopy in a highly parallel format. The electric field between parallel plates is perturbed with dielectric microstructures, generating uniform DEP forces on colloidal probes in the range of several hundred piconewtons across a macroscopic sample area. We determined the approximate crossover frequency between negative and positive DEP using electrodes without dielectric microstructures-a simplification over standard experimental methods involving quadrupoles or optical trapping. 2D and 3D simulations of the electric field distributions validated the experimental behavior of several of our DEP tweezers geometries and provided insight into potential improvements. We applied the DEP tweezers to the stretching of a short DNA oligomer and detected its extension using total-internal reflection fluorescence microscopy. The combination of a simple cell fabrication, a uniform distribution of high axial forces, and a facile optical detection of our DEP tweezers makes this form of molecular force spectroscopy ideal for highly parallel detection of stretching or unbinding kinetics of biomolecules.
我们展示了一种简单的电极几何形状在胶体探针上的应用,用于介电泳(DEP),作为一种高度平行格式的分子力谱学形式。平行板之间的电场通过介电微结构进行干扰,在宏观样品面积上产生胶体探针的均匀 DEP 力,范围在数百皮牛顿左右。我们使用没有介电微结构的电极确定了负 DEP 和正 DEP 之间的近似交叉频率——这简化了涉及四极或光阱的标准实验方法。电场分布的 2D 和 3D 模拟验证了我们的几种 DEP 镊子几何形状的实验行为,并提供了对潜在改进的见解。我们将 DEP 镊子应用于短 DNA 寡聚物的拉伸,并使用全内反射荧光显微镜检测其延伸。这种形式的分子力谱学具有简单的细胞制造、高轴向力的均匀分布以及 DEP 镊子的简便光学检测相结合,使其成为高度平行检测生物分子拉伸或解联动力学的理想选择。