论文标题
迈向定量超分辨率显微镜:具有统计保证的分子图
Towards quantitative super-resolution microscopy: Molecular maps with statistical guarantees
论文作者
论文摘要
量化荧光显微镜测量的分子数量是细胞生物学和医学研究中的重要主题。在这项工作中,我们提出了一种用于超分辨率(STED)扫描显微镜的连续算法,该算法在自动生成的图像段中提供分子计数,并以渐近置信区间的形式提供统计保证。为此,我们首先在样品的STED显微镜测量上应用多尺度扫描程序,以获得一个重要区域的系统,每个系统至少包含一个具有规定均匀概率的分子。这种区域系统通常将是高度冗余的,并由矩形构件组成。为了选择一个信息性更自然形状的区域的信息,但非冗余的子集,我们将系统杂交与通用分割算法的结果。片段的直径可以是显微镜分辨率的顺序。然后,在共焦模式下,我们使用同一样品的多个光子重合测量值,然后我们能够估算分子的亮度和数量,并在每个先前构造的段的分子计数上给出均匀的置信区间。换句话说,我们建立了一个具有统一误差控制的所谓分子图。对算法的性能进行了模拟和真实数据的研究。
Quantifying the number of molecules from fluorescence microscopy measurements is an important topic in cell biology and medical research. In this work, we present a consecutive algorithm for super-resolution (STED) scanning microscopy that provides molecule counts in automatically generated image segments and offers statistical guarantees in form of asymptotic confidence intervals. To this end, we first apply a multiscale scanning procedure on STED microscopy measurements of the sample to obtain a system of significant regions, each of which contains at least one molecule with prescribed uniform probability. This system of regions will typically be highly redundant and consists of rectangular building blocks. To choose an informative but non-redundant subset of more naturally shaped regions, we hybridize our system with the result of a generic segmentation algorithm. The diameter of the segments can be of the order of the resolution of the microscope. Using multiple photon coincidence measurements of the same sample in confocal mode, we are then able to estimate the brightness and number of the molecules and give uniform confidence intervals on the molecule counts for each previously constructed segment. In other words, we establish a so-called molecular map with uniform error control. The performance of the algorithm is investigated on simulated and real data.