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arXiv 1503.09151physics.optics
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基于超小Ge量子点的光谱超分辨荧光细胞成像

Spectroscopic super-resolution fluorescence cell imaging using ultra-small Ge quantum dots

Mingying Song, Ali Karatutlu, Osman Ersoy, Yun Zhou, Yongxin Yang, Yuanpeng Zhang, William R. Little, Ann P. Wheeler, Andrei V. Sapelkin

摘要:

在单分子定位超分辨率显微镜中,重复图像捕获的需要限制了成像速度,而荧光探针的大小限制了可能的理论定位分辨率。在这里,我们展示了一种基于光谱成像的超分辨率方法,通过在单个扫描周期内将重叠的衍射斑点分离到几个检测器中,并利用纳米颗粒中的尺寸依赖性发射波长。这种方法已经使用现成的量子点(Qdot)和内部新颖的超小(~3 nm)Ge量子点进行了测试。此外,我们开发了一种方法特定的高斯拟合和极大似然估计基于Matlab算法的快速量子点定位。我们证明,这种方法的结果在~ 40 nm的本地化分辨率使用商业量子点和~12 nm的本地化分辨率使用Ge量子点。使用标准扫描共聚焦显微镜,我们实现了1.6秒/帧的数据采集速率。然而,我们表明,这种方法有可能提供毫秒级的数据采集速率,从而在现场系统中提供超分辨率。

英文摘要:

In single molecule localisation super-resolution microscopy the need for repeated image capture limits the imaging speed, while the size of fluorescence probes limits the possible theoretical localisation resolution. Here, we demonstrated a spectral imaging based super-resolution approach by separating the overlapping diffraction spots into several detectors during a single scanning period and taking advantage of the size-dependent emission wavelength in nanoparticles. This approach has been tested using off-the-shelf quantum dots (Qdot) and in-house novel ultra-small (~3 nm) Ge QDs. Furthermore, we developed a method-specific Gaussian fitting and maximum likelihood estimation based on a Matlab algorithm for fast QDs localisation. We demonstrate that this methodology results in ~ 40 nm localisation resolution using commercial QDs and ~12 nm localisation resolution using Ge QDs. Using a standard scanning confocal microscope we achieved data acquisition rate of 1.6 seconds/frame. However, we show that this approach has a potential to deliver data acquisition rates on ms scale thus providing super-resolution in live systems.

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