实现实时重建大型视野单分子局部化显微镜,使用离散的场依赖点扩散函数
Jun Lu1, Lei Xu1, Shuyao Liao1
1Academy for Engineering and Technology, Yiwu Research Institute, Fudan University, Shanghai 200433, China.
Biomedical optics express
|February 17, 2025
概括
光学偏差限制了3D单分子定位显微镜 (SMLM) 的视野. 本研究介绍了离散的取决于现场的点分布函数 (PSF),以实现在更大区域的精确定位,改进SMLM应用程序.
科学领域:
- 生物物理学的生物物理.
- 光学显微镜的使用方法
- 超高分辨率的成像技术
背景情况:
- 单分子定位显微镜 (SMLM) 通过定位单个光分子来实现超分辨率.
- 对于SMLM必不可少的高数值光圈目标显示出偏离轴的扭曲点传播函数 (PSF),限制视野 (FOV).
- 这些异常引入了显著的3D定位错误,将SMLM限制在小FOV (例如50μm × 50μm) 上.
研究的目的:
- 系统地研究光学偏差对大型FOV 3D SMLM的影响.
- 提出和验证一种方法来扩大3D SMLM中的有效FOV.
- 为了在大型成像区域中实现实时,高精度的分子定位.
主要方法:
- 在不同的光学偏差下使用未经修改的,形的和双螺旋的PSF评估了定位准确性.
- 开发并测试了离散的场依赖PSF用于偏差校正.
- 采用GPU加速,用于模拟和生物样本的高速图像重建.
主要成果:
- 光学偏差显著降低了FOV边缘的定位精度和准确性.
- 离散的取决于场的PSF有效地纠正离轴偏差,使得在更大的区域中实现精确的3D定位.
- 用GPU加速的离散PSF模型允许实时SMLM图像重建.
结论:
- 光学偏差是大型FOV 3D SMLM的一个关键限制.
- 独立的现场依赖PSF提供了一个可行的策略来克服这些局限性,增强FOV.
- 使用GPU加速的实时重建使得大型FOV SMLM对生物研究更加实用.
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