Intensity-based axial localization approaches for multifocal plane microscopy
Intensity-based axial localization approaches for multifocal plane microscopy
复制标题
DOI：
10.1101/093013
复制
发表时间：
2016-12
期刊：
影响因子：
--
通讯作者：
Ramraj Velmurugan;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;S. Ram;E. Ward;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober
中科院分区：
文献类型：
--
作者：
Ramraj Velmurugan;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;J. Chao;S. Ram;E. Ward;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober;R. Ober
多焦平面显微镜(MUM)可以用来在大的轴向深度上对生物样品进行三维可视化，并提供在诸如单个粒子的三维跟踪和超分辨率显微镜等应用中所需的高轴向定位精度。这份报告分析了基于强度的轴向定位方法的性能，该方法应用于使用Fisher信息计算的UM数据。此外，还介绍了一种新的基于非参数强度的轴向位置估计方法--多强度查找算法(MILA)，它不同于典型的基于强度的方法，该方法利用每幅数据图像的多个强度值来确定点源的轴向位置。结果表明，MILA对成像点源的亚像素位置差异引起的潜在偏差具有较强的鲁棒性。并用实验数据对该方法的有效性进行了评估。
Multifocal plane microscopy (MUM) can be used to visualize biological samples in three dimensions over large axial depths and provides for the high axial localization accuracy that is needed in applications such as the three-dimensional tracking of single particles and superresolution microscopy. This report analyzes the performance of intensity-based axial localization approaches as applied to MUM data using Fisher information calculations. In addition, a new non-parametric intensity-based axial location estimation method, Multi-Intensity Lookup Algorithm (MILA), is introduced that, unlike typical intensity-based methods that make use of a single intensity value per data image, utilizes multiple intensity values per data image in determining the axial location of a point source. MILA is shown to be robust against potential bias induced by differences in the sub-pixel location of the imaged point source. The method's effectiveness on experimental data is also evaluated.