使用多帧运动去模糊的高通量荧光显微镜技术。
High-throughput fluorescence microscopy using multi-frame motion deblurring.
作者信息
Phillips Zachary F, Dean Sarah, Recht Benjamin, Waller Laura
机构信息
Graduate Group in Applied Science and Technology, University of California, Berkeley, Berkeley, CA 94720-1700, USA.
These authors contributed equally to this work.
出版信息
Biomed Opt Express. 2019 Dec 16;11(1):281-300. doi: 10.1364/BOE.11.000281. eCollection 2020 Jan 1.
We demonstrate multi-frame motion deblurring for gigapixel wide-field fluorescence microscopy using fast slide scanning with coded illumination. Our method illuminates the sample with multiple pulses within each exposure, in order to introduce structured motion blur. By deconvolving this known motion sequence from the set of acquired measurements, we recover the object with up to 10× higher SNR than when illuminated with a single pulse (strobed illumination), while performing acquisition at 5× higher frame-rate than a comparable stop-and-stare method. Our coded illumination sequence is optimized to maximize the reconstruction SNR. We also derive a framework for determining when coded illumination is SNR-optimal in terms of system parameters such as source illuminance, noise, and motion stage specifications. This helps system designers to choose the ideal technique for high-throughput microscopy of very large samples.
我们展示了使用带编码照明的快速玻片扫描技术，用于千兆像素宽场荧光显微镜的多帧运动去模糊。我们的方法在每次曝光期间用多个脉冲照射样本，以引入结构化运动模糊。通过从采集的测量数据集中对这个已知运动序列进行去卷积，我们恢复的物体的信噪比（SNR）比单脉冲照明（频闪照明）时高10倍，同时采集帧率比类似的凝视法高5倍。我们的编码照明序列经过优化，以最大化重建信噪比。我们还推导了一个框架，用于根据诸如光源照度、噪声和运动平台规格等系统参数，确定编码照明何时在信噪比方面是最优的。这有助于系统设计师为非常大的样本的高通量显微镜选择理想的技术。
Zotero 插件