超分辨显微镜快速STORM方法的评价

O. Ishaq, J. Elf, Carolina Wählby
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引用次数: 0

摘要

新的随机超分辨率方法的发展与荧光显微镜成像使生物过程的可视化在不断增加的空间和时间分辨率。对这类成像实验的定量评价需要能够高精度、召回地定位信号的计算分析方法。此外,希望这些方法能够快速并可能并行化,以便能够以有效的方式处理不断增加的收集数据量。本文采用基于压缩感知的方法来解决超分辨率显微镜中的信号检测问题。我们描述了先前发布的方法如何并行化,将处理时间减少至少四倍。我们还评估了贪婪优化方法在高噪声和高分子密度下的信号恢复效果。此外,我们的评估揭示了先前发表的压缩感知算法如何在高分子密度下降低到随机信号检测器的性能。最后,我们展示了成像系统点扩展函数的近似影响信号检测的召回率和精度,说明了参数优化的重要性。我们评估了不同信噪比和增加分子密度的合成数据的方法,并从活细胞的延时序列中可视化了真实超分辨率显微镜数据的性能。
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An Evaluation of the Faster STORM Method for Super-resolution Microscopy
Development of new stochastic super-resolution methods together with fluorescence microscopy imaging enables visualization of biological processes at increasing spatial and temporal resolution. Quantitative evaluation of such imaging experiments call for computational analysis methods that localize the signals with high precision and recall. Furthermore, it is desirable that the methods are fast and possible to parallelize so that the ever increasing amounts of collected data can be handled in an efficient way. We herein address signal detection in super-resolution microscopy by approaches based on compressed sensing. We describe how a previously published approach can be parallelized, reducing processing time at least four times. We also evaluate the effect of a greedy optimization approach on signal recovery at high noise and molecule density. Furthermore, our evaluation reveals how previously published compressed sensing algorithms have a performance that degrades to that of a random signal detector at high molecule density. Finally, we show the approximation of the imaging system's point spread function affects recall and precision of signal detection, illustrating the importance of parameter optimization. We evaluate the methods on synthetic data with varying signal to noise ratio and increasing molecular density, and visualize performance on real super-resolution microscopy data from a time-lapse sequence of living cells.
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