1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA. [3].
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2].
Nat Methods. 2014 Mar;11(3):313-8. doi: 10.1038/nmeth.2835. Epub 2014 Feb 2.
Super-resolution fluorescence microscopy is a powerful tool for biological research, but obtaining multiplexed images for a large number of distinct target species remains challenging. Here we use the transient binding of short fluorescently labeled oligonucleotides (DNA-PAINT, a variation of point accumulation for imaging in nanoscale topography) for simple and easy-to-implement multiplexed super-resolution imaging that achieves sub-10-nm spatial resolution in vitro on synthetic DNA structures. We also report a multiplexing approach (Exchange-PAINT) that allows sequential imaging of multiple targets using only a single dye and a single laser source. We experimentally demonstrate ten-color super-resolution imaging in vitro on synthetic DNA structures as well as four-color two-dimensional (2D) imaging and three-color 3D imaging of proteins in fixed cells.
超分辨率荧光显微镜是生物学研究的有力工具，但获取大量不同目标物种的多路复用图像仍然具有挑战性。在这里，我们使用短荧光标记寡核苷酸（DNA-PAINT，点积累成像在纳米级形貌中的一种变体）的瞬时结合，实现了简单易用的多路复用超分辨率成像，在体外合成 DNA 结构上实现了小于 10nm 的空间分辨率。我们还报告了一种多路复用方法（Exchange-PAINT），该方法仅使用单个染料和单个激光源即可对多个目标进行顺序成像。我们在体外合成 DNA 结构上进行了十色超分辨率成像实验，并在固定细胞中进行了四色二维（2D）成像和三色 3D 成像。
