Abstract
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 ∼200 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.
Highlights
Nanoscale correlation of structural information acquisition with specific-molecule identification provides new insight for studying rare subcellular events
The spacing of the raster scanning and the interval of the signal used to trigger camera image recording during lateral scanning are adjusted based on the area of the field of view (FOV) of microsphere superlenses without apparent aberration (Fig. 1b and Supplementary Fig. 2)
Because these filaments are not distributed across the cell membrane, they should exist inside the cell, which is similar to the results shown in Figs 3l and 4b,i; sub-membrane nanostructures that reside inside live cells (Fig. 5f) or fixed cells (Fig. 5h) beyond the diffraction barrier can be non-invasively explored over a large area under white-light illumination
Summary
Nanoscale correlation of structural information acquisition with specific-molecule identification provides new insight for studying rare subcellular events. 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. The FOV increases linearly with the diameter of the superlens, the resolving ability deteriorates as the diameter increases[23] This limited FOV reduces the range of practical applications; methods, such as probe-based manipulations[18,21] and immersing microspheres into elastomers[19,20,28], have been reported to locate microsphere superlenses for observations from a specific position. Compared with point-based raster scanning microscopes, this new technique replaces the ‘point’ with an ‘area’ (that is, the middle area of the FOV of the microsphere superlens has no aberrations); there is a significant improvement in terms of the time efficiency for large-area imaging
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
