Real-time high dynamic range laser scanning microscopy.
Real-time high dynamic range laser scanning microscopy.
复制标题
实时高动态范围激光扫描显微镜。
DOI：
10.1038/ncomms11077
复制
发表时间：
2016-04-01
影响因子：
16.6
通讯作者：
Weissleder R
中科院分区：
文献类型：
--
作者：
Vinegoni C;Leon Swisher C;Fumene Feruglio P;Giedt RJ;Rousso DL;Stapleton S;Weissleder R
在传统的共焦/多光子荧光显微镜中，图像通常是在理想的设置下并在对给定结构或特征的参数进行广泛优化之后获得的，这通常导致来自其他图像属性的信息丢失。为了克服选择性数据显示的问题，我们开发了一种新的方法，扩大了光学显微镜的成像动态范围，提高了信噪比。在这里，我们展示了实时和连续的高动态范围显微镜如何促进自动化的三维神经分割。我们解决了不同大小、解剖结构和复杂性的样本的重建和分割性能。最后，还展示了活体实时高动态范围成像，使该技术特别适用于存在生理运动的纵向成像和/或功能成像期间体内快速示踪剂动力学的量化。共焦和多光子荧光显微镜通常存在动态范围小的问题。在这里，作者开发了一种高动态范围的激光扫描荧光技术，通过同时记录不同的光强范围。该方法可适用于商业系统。
In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging. Confocal and multiphoton fluorescence microscopy often suffers from low dynamic range. Here the authors develop a high dynamic range, laser scanning fluorescence technique by simultaneously recording different light intensity ranges. The method can be adapted to commercial systems.