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Scanning Superlens Microscopy for Non-Invasive Large Field-Of-View Visible Light Nanoscale Imaging

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Scanning Superlens Microscopy for Non-Invasive Large Field-Of-View Visible Light Nanoscale Imaging Scanning superlens microscopy for non-invasive large field-of-view visible light nanoscale imaging Wang, Feifei; Liu, Lianqing; Yu, Haibo; Wen, Yangdong; Yu, Peng; Liu, Zhu; Wang, Yuechao; Li, Wen Jung Published in: Nature Communications Published: 09/12/2016 Document Version: Final Published version, also known as Publisher’s PDF, Publisher’s Final version or Version of Record License: CC BY Publication record in CityU Scholars: Go to record Published version (DOI): 10.1038/ncomms13748 Publication details: Wang, F., Liu, L., Yu, H., Wen, Y., Yu, P., Liu, Z., Wang, Y., & Li, W. J. (2016). Scanning superlens microscopy for non-invasive large field-of-view visible light nanoscale imaging. Nature Communications, 7, [13748]. https://doi.org/10.1038/ncomms13748 Citing this paper Please note that where the full-text provided on CityU Scholars is the Post-print version (also known as Accepted Author Manuscript, Peer-reviewed or Author Final version), it may differ from the Final Published version. When citing, ensure that you check and use the publisher's definitive version for pagination and other details. General rights Copyright for the publications made accessible via the CityU Scholars portal is retained by the author(s) and/or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights. Users may not further distribute the material or use it for any profit-making activity or commercial gain. Publisher permission Permission for previously published items are in accordance with publisher's copyright policies sourced from the SHERPA RoMEO database. Links to full text versions (either Published or Post-print) are only available if corresponding publishers allow open access. Take down policy Contact [email protected] if you believe that this document breaches copyright and provide us with details. We will remove access to the work immediately and investigate your claim. Download date: 23/09/2021 ARTICLE Received 15 Dec 2015 | Accepted 31 Oct 2016 | Published 9 Dec 2016 DOI: 10.1038/ncomms13748 OPEN Scanning superlens microscopy for non-invasive large field-of-view visible light nanoscale imaging Feifei Wang1,2, Lianqing Liu1, Haibo Yu1, Yangdong Wen1,2, Peng Yu1, Zhu Liu1, Yuechao Wang1 & Wen Jung Li1,3 Nanoscale correlation of structural information acquisition with specific-molecule identification provides new insight for studying rare subcellular events. To achieve this correlation, scanning electron microscopy has been combined with super-resolution fluorescent microscopy, despite its destructivity when acquiring biological structure information. Here we propose time-efficient non-invasive microsphere-based scanning superlens microscopy that enables the large-area observation of live-cell morphology or sub-membrane structures with sub-diffraction-limited resolution and is demonstrated by observing biological and non-biological objects. This microscopy operates in both non-invasive and contact modes with B200 times the acquisition efficiency of atomic force microscopy, which is achieved by replacing the point of an atomic force microscope tip with an imaging area of microspheres and stitching the areas recorded during scanning, enabling sub-diffraction-limited resolution. Our method marks a possible path to non-invasive cell imaging and simultaneous tracking of specific molecules with nanoscale resolution, facilitating the study of subcellular events over a total cell period. 1 State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016, China. 2 University of Chinese Academy of Sciences, Beijing 100049, China. 3 Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Kowloon Tong 999077, Hong Kong. Correspondence and requests for materials should be addressed to L.L. (email: [email protected]) or to W.J.L. (email: [email protected]). NATURE COMMUNICATIONS | 7:13748 | DOI: 10.1038/ncomms13748 | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms13748 o fully explore the basic mechanisms in life and material FOV increases linearly with the diameter of the superlens, the sciences, and in other areas, research has gradually resolving ability deteriorates as the diameter increases23. This Tmoved into the nanoscale using various novel microscopy limited FOV reduces the range of practical applications; therefore, technologies. Traditional electron microscopy, which achieves methods, such as probe-based manipulations18,21 and immersing a molecular- and atomic-level resolution, has played an microspheres into elastomers19,20,28, have been reported to locate irreplaceable role in this transition; however, the radiation dose microsphere superlenses for observations from a specific position. that the specimens are exposed to by the imaging system’s Unfortunately, when using these methods, it is difficult to achieve energetic electron beam is comparable to the irradiation from the large-area imaging and to locate specific sub-diffraction-limited explosion of a 10-megaton hydrogen bomb B30 m away, targets in a large area. In particular, these targets appear in which confines the application of electron microscopy to non- different areas, such as the distribution of lysosomes in cells. destructive cell observation1,2. Scanning probe microscopies In addition to these methods, microspheres or introduced provide another convenient approach to obtain sub-nanometre elastomers still make contact with the samples, and the distance resolution in both air and liquid, but their invasiveness may or interaction force between the microspheres and samples induce unrecoverable structural damage or morphology cannot be precisely controlled due to the lack of effective feedback deformation in sensitive specimens. Specifically, the interaction mechanisms, which further adds to the invasiveness of this forces they produce may drive adherent cells to detach from a microsphere-based super-resolution imaging method. dish surface during scanning with atomic force microscopy In the following, we introduce a non-invasive, environmentally (AFM). The adhesion between cells and tips causes cell damage compatible and high-throughput optical microscopy technique and limits the reusability of the probe and potentially contributes called scanning superlens microscopy (SSUM) for large-area, to cross-contamination. Recently, advancements in life studies super-resolution imaging and structural information acquisition. have accelerated markedly due to the development of super- The distance or interaction forces between the microsphere resolution fluorescence microscopes that circumvent the optical superlenses and the specimens are adjusted using AFM principles, diffraction limit induced by the loss of evanescent waves in far- allowing the scanning and imaging of large areas. On the basis of field imaging, producing nanoscale resolutions3–5. Fluorescent the microsphere superlens imaging properties, two different AFM labelling could allow the identification of specific molecules scanning modes that are compatible with microsphere superlens and to explore the functions of these molecules in life4,6; however, imaging have been demonstrated to achieve microsphere this labelling also induces a loss of structural information because scanning of a large area: the contact mode enables imaging a majority of molecules are unlabeled7. The implementation of stiff samples, and the constant-height mode facilitates of a large fraction of these imaging methods depends on a non-invasive imaging of sensitive specimens. Compared with point-based raster scanning technology, such as the commonly point-based raster scanning microscopes, this new technique used scanning electron microscopy (SEM)8, scanning replaces the ‘point’ with an ‘area’ (that is, the middle area of the probe microscopy (SPM)9,10 and stimulated emission depletion FOV of the microsphere superlens has no aberrations); therefore, (STED)11 microscopy. The acquisition time increases significantly there is a significant improvement in terms of the time efficiency as the imaging area (for example, 450 Â 50 mm2) is enlarged at a for large-area imaging. high resolution (for example, o100 nm). In addition, SPM and SEM typically use a slow scanning speed and increase the number of scan lines used to improve imaging quality12, which further Results leads to time inefficiencies. To increase the imaging speed, Description of the microsphere-based SSUM. Figure 1a shows parallelized scanning systems have been designed for different the basic concept of a microsphere-based SSUM. An AFM types of microscopes (for example, SEM13, SPM14,15 and distance or interaction force control mechanism is introduced to STED16,17), making the systems more complex and expensive, adjust the position of the microsphere above the sample. and requiring sophisticated algorithms to achieve system The magnification of the microscope objectives used in the control and image processing18. Only few of these systems microsphere superlens-based nanoscale imaging is typically are commercially available currently. Compared with these 4 Â 50 with a limited working distance18,22–26. To be compatible microscopes, traditional optical microscopes have certain with these objectives, a custom AFM system (Supplementary unique advantages, such as the ability for non-invasive, real- Fig. 1) illuminates and collects the laser beam of the optical lever time, large-area and fluorescent
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