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arXiv 1704.04091physics.opticsphysics.bio-ph
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环形照明的高分辨率强度传输定量相位显微术

High-resolution transport-of-intensity quantitative phase microscopy with annular illumination

Chao Zuo, Jiasong Sun, Jiaji Li, Jialin Zhang, Anand Asundi, Qian Chen

摘要:

对于基于强度输运方程(TIE)的定量相位成像(QPI),部分相干照明提供无散斑成像、与明场显微镜兼容以及超过相干衍射极限的横向分辨率。不幸的是,在具有圆形照明孔径的传统显微镜中,部分相干性往往会减小相位对比度,加剧TIE成像中固有的噪声-分辨率权衡,从而导致强烈的低频伪影和受损的成像分辨率。在这里,我们演示了如何可以有效地解决这些问题,取代传统的圆形照明孔径与环形。匹配环形照明不仅大大提高了低空间频率的相位对比度,而且显著提高了实际成像分辨率,接近非相干衍射极限。通过将高数值孔径(NA)照明以及高NA物镜,它示出,第一次,TIE相位成像可以实现高达208 nm的横向分辨率,对应于2.66的有效NA。举例说明了在体外Hela细胞的延时成像,揭示了细胞有丝分裂和凋亡过程中的细胞形态和亚细胞动力学。鉴于其高分辨率QPI的能力以及与广泛使用的明场显微镜硬件的兼容性,预计所提出的方法将被更广泛的生物学和医学界采用。

英文摘要:

For quantitative phase imaging (QPI) based on transport-of-intensity equation (TIE), partially coherent illumination provides speckle-free imaging, compatibility with brightfield microscopy, and transverse resolution beyond coherent diffraction limit. Unfortunately, in a conventional microscope with circular illumination aperture, partial coherence tends to diminish the phase contrast, exacerbating the inherent noise-to-resolution tradeoff in TIE imaging, resulting in strong low-frequency artifacts and compromised imaging resolution. Here, we demonstrate how these issues can be effectively addressed by replacing the conventional circular illumination aperture with an annular one. The matched annular illumination not only strongly boosts the phase contrast for low spatial frequencies, but significantly improves the practical imaging resolution to near the incoherent diffraction limit. By incorporating high-numerical aperture (NA) illumination as well as high-NA objective, it is shown, for the first time, that TIE phase imaging can achieve a transverse resolution up to 208 nm, corresponding to an effective NA of 2.66. Time-lapse imaging of in vitro Hela cells revealing cellular morphology and subcellular dynamics during cells mitosis and apoptosis is exemplified. Given its capability for high-resolution QPI as well as the compatibility with widely available brightfield microscopy hardware, the proposed approach is expected to be adopted by the wider biology and medicine community.

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