Ames Laboratory, U.S. Department of Energy, and Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
J Am Chem Soc. 2012 Apr 11;134(14):6108-11. doi: 10.1021/ja301332t. Epub 2012 Apr 2.
Novel non-blinking quantum dots (NBQDs) were utilized in three-dimensional super-localization, high-precision tracking applications under an automated scanning-angle total internal reflection fluorescence microscope (SA-TIRFM). NBQDs were randomly attached to stationary microtubules along the radial axis under gliding assay conditions. By automatically scanning through a wide range of incident angles with different evanescent-field layer thicknesses, the fluorescence intensity decay curves were obtained. Fit with theoretical decay functions, the absolute vertical positions were determined with sub-10-nm localization precision. The emission intensity profile of the NBQDs attached to kinesin-propelled microtubules was used to resolve the self-rotation of gliding microtubules within a small vertical distance of ~50 nm. We demonstrate the applicability of NBQDs in high-precision fluorescence imaging experiments.
新型无闪烁量子点 (NBQDs) 被用于在自动扫描角全内反射荧光显微镜 (SA-TIRFM) 下进行三维超定位、高精度跟踪应用。在滑动实验条件下，NBQDs 随机附着在沿径向的固定微管上。通过自动扫描具有不同消逝场层厚度的宽入射角范围，获得荧光强度衰减曲线。通过与理论衰减函数拟合，确定了具有亚 10nm 定位精度的绝对垂直位置。附着在驱动蛋白推动的微管上的 NBQDs 的发射强度分布用于解析在小垂直距离 (~50nm) 内滑行微管的自旋转。我们展示了 NBQDs 在高精度荧光成像实验中的适用性。
