School of Biology, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan, China 410082.
Anal Chem. 2012 Feb 7;84(3):1504-9. doi: 10.1021/ac202784h. Epub 2012 Jan 24.
The key factor of realizing super-resolution optical microscopy at the single-molecule level is to separately position two adjacent molecules. An opportunity to independently localize target molecules is provided by the intermittency (blinking) in fluorescence of a quantum dot (QD) under the condition that the blinking of each emitter can be recorded and identified. Herein we develop a spectral imaging based color nanoscopy which is capable of determining which QD is blinking in the multicolor QD complex through tracking the first-order spectrum, and thus, the distance at tens of nanometers between two QDs is measured. Three complementary oligonucleotides with lengths of 15, 30, and 45 bp are constructed as calibration rulers. QD585 and QD655 are each linked at one end. The measured average distances are in good agreement with the calculated lengths with a precision of 6 nm, and the intracellular dual-color QDs within a diffraction-limited spot are distinguished.
实现单分子水平超分辨光学显微镜的关键因素是分别定位两个相邻分子。在每个发射器的闪烁都可以被记录和识别的情况下，量子点（QD）荧光的间歇（闪烁）为目标分子的独立定位提供了机会。本文中我们开发了一种基于光谱成像的彩色纳米显微镜，通过跟踪一阶光谱，确定在多色 QD 复合物中哪个 QD 正在闪烁，从而测量两个 QD 之间数十纳米的距离。构建了三个长度为 15、30 和 45 bp 的互补寡核苷酸作为校准标尺。QD585 和 QD655 分别在一端连接。测量的平均距离与计算长度吻合良好，精度为 6nm，并且可以分辨出在一个衍射极限点内的细胞内双色 QD。
