Phase-retrieved pupil functions in wide-field fluorescence microscopy
Phase-retrieved pupil functions in wide-field fluorescence microscopy
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DOI：
10.1111/j.0022-2720.2004.01393.x
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发表时间：
2004-10-01
影响因子：
2
通讯作者：
Sedat, JW
中科院分区：
文献类型：
--
作者：
Hanser, BM;Gustafsson, MGL;Sedat, JW
光瞳函数是对宽视场光学系统三维成像特性的紧凑且可修改的描述。可以使用相位恢复算法从测量的点扩散函数（PSF）计算地估计显微镜的光瞳函数。将3D PSF压缩到2D光瞳函数中抑制了伪影和测量噪声，而无需求助于旋转平均。我们在这里表明，这样的“相位检索”的瞳孔功能可以再现的光路中的功能，无论是在样品附近，并在显微镜。与PSF不同，光瞳函数可以简单地通过将光瞳函数乘以计算的像差函数而容易地修改为包括已知像差，诸如由折射率失配的安装介质引起的那些像差。从这样的修改的瞳孔函数计算的PSF紧密地匹配在像差成像条件下收集的对应的测量的PSF。当用于模拟对象的图像去卷积时，这些相位恢复的计算的PSF与直接测量的PSF类似地执行。
Pupil functions are compact and modifiable descriptions of the three-dimensional (3D) imaging properties of wide-field optical systems. The pupil function of a microscope can be computationally estimated from the measured point spread function (PSF) using phase retrieval algorithms. The compaction of a 3D PSF into a 2D pupil function suppresses artefacts and measurement noise without resorting to rotational averaging. We show here that such 'phase-retrieved' pupil functions can reproduce features in the optical path, both near the sample and in the microscope. Unlike the PSF, the pupil function can be easily modified to include known aberrations, such as those induced by index-mismatched mounting media, simply by multiplying the pupil function by a calculated aberration function. PSFs calculated from such a modified pupil function closely match the corresponding measured PSFs collected under the aberrated imaging conditions. When used for image deconvolution of simulated objects, these phase-retrieved, calculated PSFs perform similarly to directly measured PSFs.