论文标题
时间调节的激发，以增强单分子定位显微镜
Time-modulated excitation for enhanced single molecule localization microscopy
论文作者
论文摘要
单分子定位显微镜中的结构化照明提供了有关分子位置的新信息，从而提高了与标准定位方法相比的定位精度。在这里，我们使用时间班的正弦激发模式来调节分子的荧光信号，其位置信息是通过相通过同步解调恢复的。我们设计了两个灵活的快速解调系统，位于相机上游，使我们能够克服限制摄像机的采集频率，从而最大程度地收集了在解调过程中的光子收集。然后在采集期间反复在同一图像上同步对时间调制的荧光信号同步采样。与经典的高斯拟合方法相比，这种称为MODLOC的显微镜（称为MODLOC）可以在一个方向上提高定位精度为2.4。一项时间研究和实验性证明都表明，闪烁的分子的寿命很短，在Kilohertz范围内施加了一个调制频率，这超出了当前摄像机的范围。因此，必须在这些频率上运行的解调系统充分利用这种新的本地化方法。
Structured illumination in Single Molecule Localization Microscopy provides new information on the position of molecules and thus improves the localization precision compared to standard localization methods. Here, we used a time-shifted sinusoidal excitation pattern to modulate the fluorescence signal of the molecules whose position information is carried by the phase and recovered by synchronous demodulation. We designed two flexible fast demodulation systems located upstream of the camera, allowing us to overcome the limiting camera acquisition frequency and thus to maximize the collection of photons in the demodulation process. The temporally modulated fluorescence signal was then sampled synchronously on the same image, repeatedly during acquisition. This microscopy, called ModLoc, allows to experimentally improve the localization precision by a factor of 2.4 in one direction, compared to classical Gaussian fitting methods. A temporal study and an experimental demonstration both show that the short lifetimes of the molecules in blinking regimes impose a modulation frequency in the kilohertz range, which is beyond the reach of current cameras. A demodulation system operating at these frequencies would thus be necessary to take full advantage of this new localization approach.
