Theoretical Limits on Errors and Acquisition Rates in Localizing Switchable Fluorophores
Theoretical Limits on Errors and Acquisition Rates in Localizing Switchable Fluorophores
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DOI：
10.1016/j.bpj.2008.11.001
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发表时间：
2009-01-21
影响因子：
3.4
通讯作者：
Small, Alexander R.
中科院分区：
文献类型：
--
作者：
Small, Alexander R.
最近的各种成像技术能够通过激活和定位标本中荧光分子的子集，并重复这一过程，直到所有分子都被成像，从而在荧光显微复制中突破衍射极限。在这些技术中，需要在速度（每个成像周期激活更多分子）和错误率（激活更多分子有可能产生隐藏分子位置信息的重叠图像）之间进行权衡，因此需要智能图像处理方法来识别和拒绝重叠图像。我们在这里介绍了一个定义错误率的形式，推导了错误率、图像采集率和图像处理算法性能特征之间的一般关系，并表明无论算法性能如何，都存在最小的采集时间。我们还考虑了从重叠模糊图像中推断分子位置的算法，并推导了最小获取时间对算法性能的依赖关系。
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.