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Theoretical Limits on Errors and Acquisition Rates in Localizing Switchable Fluorophores

机译:局域可转换荧光团的误差和捕获率的理论极限

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摘要

A variety of recent imaging techniques are able to beat the diffraction limit in fluorescence microcopy by activating and localizing subsets of the fluorescent molecules in the specimen, and repeating this process until all of the molecules have been imaged. In these techniques there is a tradeoff between speed (activating more molecules per imaging cycle) and error rates (activating more molecules risks producing overlapping images that hide information on molecular positions), and so intelligent image processing approaches are needed to identify and reject overlapping images. We introduce here a formalism for defining error rates, derive a general relationship between error rates, image acquisition rates, and the performance characteristics of the image processing algorithms, and show that there is a minimum acquisition time irrespective of algorithm performance. We also consider algorithms that can infer molecular positions from images of overlapping blurs, and derive the dependence of the minimum acquisition time on algorithm performance.
机译:通过激活和定位样本中荧光分子的子集,并重复此过程,直到所有分子都已成像,各种最新的成像技术都能够突破荧光显微镜的衍射极限。在这些技术中,需要在速度(每个成像周期激活更多的分子)和错误率(激活更多的分子可能产生重叠的图像,从而在分子位置上隐藏信息)之间权衡取舍,因此需要智能图像处理方法来识别和拒绝重叠的图像。我们在这里介绍一种用于定义错误率的形式,推导错误率,图像采集率和图像处理算法的性能特征之间的一般关系,并表明与算法性能无关的最小采集时间。我们还考虑了可以从重叠的模糊图像推断分子位置的算法,并得出最小采集时间对算法性能的依赖性。

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