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Space non-invariant point-spread function and its estimation in fluorescence microscopy

机译:空间不变点扩散函数及其在荧光显微镜中的估计

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摘要

In this research report, we recall briefly how the diffraction-limited nature of an optical microscope's objective, and the intrinsic noise can affect the observed images' resolution. A blind deconvolution algorithm can restore the lost frequencies beyond the diffraction limit. However, under other imaging conditions, the approximation of aberration-free imaging, is not applicable, and the phase aberrations of the emerging wavefront from a specimen immersion medium cannot be ignored any more. We show that an object's location and its original intensity distribution can be recovered by retrieving the refracted wavefront's phase from the observed intensity images. We demonstrate this by retrieving the point-spread function from an imaged microsphere. The noise and the influence of the microsphere size can be mitigated and sometimes completely removed from the observed images by using a maximum a posteriori estimate. However, due to the incoherent nature of the acquisition system, phase retrieval from the observed intensities will be possible only if the phase is constrained. We have used geometrical optics to model the phase of the refracted wavefront, and tested the algorithm on some simulated images.
机译:在本研究报告中,我们简要回顾了光学显微镜物镜的衍射极限特性和固有噪声如何影响观察到的图像的分辨率。盲解卷积算法可以将丢失的频率恢复到衍射极限之外。然而,在其他成像条件下,无像差成像的近似是不适用的,并且从样品浸没介质中出现的波前的相位像差不能再被忽略了。我们表明,可以通过从观察到的强度图像中检索折射波前的相位来恢复对象的位置及其原始强度分布。我们通过从成像的微球检索点扩展函数来证明这一点。通过使用最大后验估计,可以减轻噪声和微球尺寸的影响,有时可以将其从观察到的图像中完全去除。但是,由于采集系统的不连贯性,只有在相位受到约束的情况下,才有可能从观察到的强度中进行相位检索。我们已经使用几何光学对折射波前的相位进行建模,并在一些模拟图像上测试了该算法。

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