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远场无标记超分辨率成像的新方法。

Novel approach for label free super-resolution imaging in far field.

作者信息

Alexandrov Sergey A, McGrath James, Subhash Hrebesh, Boccafoschi Francesca, Giannini Cinzia, Leahy Martin

机构信息

Tissue Optics &Microcirculation Imaging Group, School of Physics, National University of Ireland, Galway, Ireland.

Department of Health Sciences, University of Piemonte Orientale "A. Avogadro", 28100 Novara, Italy.

出版信息

Sci Rep. 2015 Sep 3;5:13274. doi: 10.1038/srep13274.

Abstract

Progress in the emerging areas of science and technology, such as bio- and nano-technologies, depends on development of corresponding techniques for imaging and probing the structures with high resolution. Recently, the far field diffraction resolution limit in the optical range has been circumvented and different methods of super-resolution optical microscopy have been developed. The importance of this breakthrough achievement has been recognized by Nobel Prize for Chemistry in 2014. However, the fluorescence based super-resolution techniques only function with fluorescent molecules (most of which are toxic and can destroy or lead to artificial results in living biological objects) and suffer from photobleaching. Here we show a new way to break the diffraction resolution limit, which is based on nano-sensitivity to internal structure. Instead of conventional image formation as 2D intensity distribution, in our approach images are formed as a result of comparison of the axial spatial frequency profiles, reconstructed for each image point. The proposed approach dramatically increases the lateral resolution even in presence of noise and allows objects to be imaged in their natural state, without any labels.

摘要

诸如生物和纳米技术等新兴科技领域的进展,取决于开发相应的高分辨率成像和探测结构的技术。最近,光学范围内的远场衍射分辨率极限已被突破,并且已经开发出不同的超分辨率光学显微镜方法。这一突破性成就的重要性已在2014年获得诺贝尔化学奖认可。然而,基于荧光的超分辨率技术仅对荧光分子起作用(其中大多数有毒,会破坏活的生物物体或导致人为结果),并且存在光漂白问题。在此,我们展示了一种突破衍射分辨率极限的新方法,该方法基于对内部结构的纳米敏感性。与传统的作为二维强度分布的图像形成方式不同,在我们的方法中,图像是通过比较为每个图像点重建的轴向空间频率分布而形成的。所提出的方法即使在存在噪声的情况下也能显著提高横向分辨率,并允许在物体处于自然状态下成像,无需任何标记。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e628/4558609/1c3801766bc9/srep13274-f1.jpg

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