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Xiao YT, Chen LW, Pu MB et al. Improved spatiotemporal resolution of anti-scattering super-resolution label-free microscopy via synthetic wave 3D metalens imaging. Opto-Electron Sci 2, 230037 (2023). doi: 10.29026/oes.2023.230037
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Article Open Access
Improved spatiotemporal resolution of anti-scattering super-resolution label-free microscopy via synthetic wave 3D metalens imaging
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Abstract
Super-resolution (SR) microscopy has dramatically enhanced our understanding of biological processes. However, scattering media in thick specimens severely limits the spatial resolution, often rendering the images unclear or indistinguishable. Additionally, live-cell imaging faces challenges in achieving high temporal resolution for fast-moving subcellular structures. Here, we present the principles of a synthetic wave microscopy (SWM) to extract three-dimensional information from thick unlabeled specimens, where photobleaching and phototoxicity are avoided. SWM exploits multiple-wave interferometry to reveal the specimen’s phase information in the area of interest, which is not affected by the scattering media in the optical path. SWM achieves ~0.42λ/NAresolution at an imaging speed of up to 106 pixels/s. SWM proves better temporal resolution and sensitivity than the most conventional microscopes currently available while maintaining exceptional SR and anti-scattering capabilities. Penetrating through the scattering media is challenging for conventional imaging techniques. Remarkably, SWM retains its efficacy even in conditions of low signal-to-noise ratios. It facilitates the visualization of dynamic subcellular structures in live cells, encompassing tubular endoplasmic reticulum (ER), lipid droplets, mitochondria, and lysosomes.-
Keywords:
- super-resolution /
- anti-scattering /
- unlabeled /
- high temporal resolution
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References
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Xiao YT, Chen LW, Pu MB et al. Improved spatiotemporal resolution of anti-scattering super-resolution label-free microscopy via synthetic wave 3D metalens imaging. Opto-Electron Sci 2, 230037 (2023). doi: 10.29026/oes.2023.230037
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Figure 1.
(a) The SWM system configuration comprises four main components: a synthetic wave source generation module, a metalens, a pinhole, and a PMT. (b) The schematic diagram of the post-processing process. The first row is the four phase-shifting frames. The second row is the reconstructed quantitative phase gradient image. The third row is the reconstructed quantitative phase image.
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Figure 2.
(a) The unit cell of the metalens is shown from both a top view and a side view. (b) The transmission coefficient's amplitude and (c) its phase delay for square lattice periodic nanopillar transmit arrays as a function of duty cycles of the nanopillars and the lattice constant. The square lattice period of 500 nm is indicated by a white dashed line. (d) The amplitude and phase of the transmission coefficient with a fixed lattice constant of 500 nm while varying the width of the nanopillars. (e) The calculated phase profile. (f) The schematic of metalens operating in transmission mode.
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Figure 3.
(a) Normalized intensity distribution when metawave source is incident normally. The first row is 2D distribution. The dotted line indicates the Airy spot area. The second row is 1D distribution along the x- and y-direction. The arrow indicates the FWHM of focus spot. (b) Lateral resolution as a function of incident angles. (c) Normalized intensity profiles of the focal spot for different incident angles x-y plane. (d) Normalized intensity distribution along the x-direction in the x-y plane when metawave source is incident with different angles. The arrow represents the resolution of the SWM. The incident angles are 2.5°, 2°, 1.9° and 1.8°, respectively. D represents the distance between the centers of two focused light spots.
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Figure 4.
(a) Normalized intensity curve recorded by PMT during one period for different phase differences. (b) The top section represents an anti-scattering imaging model with a medium screen positioned above the imaged object, while the bottom section depicts an anti-scattering imaging model with a medium screen placed below the imaged object. On the left-hand side, there is a map illustrating the distribution of the scattering media's phase. (c) Reconstruction phase difference vs. real phase difference. (d) Detection error as it varies with the distance of the scattering medium screen from the metalens, with the medium screen located above the imaged object. (e) Detection error as it varies with the distance of the scattering medium screen from the sample, with the medium screen situated below the imaged object.
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Figure 5.
(a) LS-GLIM image and SWM image taken without the scattering layer. (b) traditional image taken with the scattering layer. (c) SWM image taken with the scattering layer.
