用于SMLM定位和显微镜表征的点差函数的通用反向建模
Sheng Liu1, Jianwei Chen2,3, Jonas Hellgoth4
1Department of Physics and Astronomy, University of New Mexico, Albuquerque, NM, USA.
bioRxiv : the preprint server for biology
|November 14, 2023
概括
精确的显微镜点扩散函数 (PSF) 模型对于超高分辨率成像至关重要. uiPSF工具箱从显微镜数据中推断出精确的PSF模型,使先进的3D超分辨率成像和系统表征成为可能.
科学领域:
- 显微镜的使用方法
- 光学成像技术的成像
- 计算生物学 计算生物学
背景情况:
- 点扩散函数 (PSF) 模拟了显微镜如何成像点源,这对于图像分析至关重要.
- 准确的PSF模型对于单分子局部化显微镜 (SMLM),解卷和偏差校正至关重要.
- 现有的方法可能缺乏普遍性或需要特定的数据类型来推断PSF.
研究的目的:
- 为从显微镜数据中推断点传播函数 (PSF) 模型开发一种通用和准确的工具箱.
- 为了实现高精度的3D超分辨率成像和显微镜系统的详细表征.
- 提供适用于不同显微镜模式和数据源的灵活框架.
主要方法:
- 介绍了 uiPSF (点差函数的通用反向建模),用于PSF模型推断的工具箱.
- 使用光珠的图像堆或闪的光灯的直接图像 (SMLM原始数据).
- 开发了一个模块化框架,结合了系统特定的特征,如珠子大小和通道转换.
主要成果:
- 成功推断出准确的PSF模型,通过SMLM实现高保真度3D超分辨率成像.
- 证明了显微镜误差的量化,包括取决于场的误差,以及分辨率.
- 在各种SMLM系统 (4Pi-SMLM,格子光片) 和数据类型 (珠子,单个分子) 中验证了 uiPSF.
结论:
- uiPSF工具箱为显微镜中PSF模型推理提供了一个通用和准确的方法.
- 精确的PSF建模显著提高了3D超分辨率成像能力和显微镜表征.
- 该框架的模块化性和适用于各种系统的可用性使其成为光学成像研究的宝贵工具.
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