An entropy-controlled objective chip for reflective confocal microscopy with subdiffraction-limit resolution

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: Sept. 20, 2023

Planar diffractive lenses (PDLs) with optimized but disordered structures can focus light beyond the diffraction limit. However, these have inevitably destroyed wide-field imaging capability, limiting their applications in microscopy. Here, we introduce information entropy S to evaluate disorder of an objective chip by using probability its structural deviation from standard Fresnel zone plates. Inspired theory change, predict equilibrium point [Formula: see text] balance (theoretically evaluated Strehl ratio) and subdiffraction-limit focusing. To verify this, a record-long focal length 1 mm is designed text], which nearest among all reported PDLs. Consequently, our fabricated size 0.44 λ image fine details spatial frequencies up 4000 lp/mm experimentally. These unprecedented performances enable ultracompact reflective confocal microscopy for superresolution imaging.

Language: Английский

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MINFLUX Nanoscopy: A “Brilliant” Technique Promising Major Breakthrough

Microscopy Research and Technique, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 21, 2025

ABSTRACT MINFLUX nanoscopy relies on the localization of single fluorophores with expected ~ 2 nm precision in 3D mapping, roughly one order magnitude better than standard stimulated emission depletion microscopy or stochastic optical reconstruction microscopy. This “brilliant” technique takes advantage specialized principles and algorithms that require only dim fluorescence signals a minimum flux photons; hence name follows. With this level performance, imaging tracking should allow for routine study biological processes down to molecular scale, revealing previously unresolved details cell structures, such as organization calcium channels muscle cells clustering receptors synapses. Whereas high is definitely strength technique, limitations challenges also exist, especially labeling procedures aiming at appropriate density on/off switching kinetics. primer presents some significant results achieved so far highlights specific operational crucial technique.

Language: Английский

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Deep learning enhanced light sheet fluorescence microscopy for in vivo 4D imaging of zebrafish heart beating

Light Science & Applications, Journal Year: 2025, Volume and Issue: 14(1)

Published: Feb. 25, 2025

Abstract Time-resolved volumetric fluorescence imaging over an extended duration with high spatial/temporal resolution is a key driving force in biomedical research for investigating spatial-temporal dynamics at organism-level systems, yet it remains major challenge due to the trade-off among speed, light exposure, illumination power, and image quality. Here, we present deep-learning enhanced sheet microscopy (LSFM) approach that addresses restoration of rapid time-lapse less than 0.03% exposure 3.3% acquisition time compared typical standard acquisition. We demonstrate convolutional neural network (CNN)-transformer developed here, namely U-net integrated transformer (UI-Trans), successfully achieves mitigation complex noise-scattering-coupled degradation outperforms state-of-the-art deep learning networks, its capability faithfully fine details while comprehending global features. With fast generation appropriate training data via flexible switching between confocal line-scanning LSFM (LS-LSFM) conventional LSFM, this method three- five-fold signal-to-noise ratio (SNR) improvement ~1.8 times contrast ex vivo zebrafish heart long-term 4D (3D morphology + time) heartbeat different developmental stages ultra-economical acquisitions terms dosage time.

Language: Английский

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Guidelines for MINFLUX Excitation Pattern Design

ACS Photonics, Journal Year: 2025, Volume and Issue: unknown

Published: March 9, 2025

Language: Английский

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Deep-Penetrating and High-Resolution Continuous-Wave Nonlinear Microscopy Based on Homologous Dual-Emission Upconversion Adaptive Optics

Nano Letters, Journal Year: 2025, Volume and Issue: unknown

Published: March 20, 2025

Lanthanide-doped upconversion nanoparticles (UCNPs) are emerging as innovative nonlinear probes in biomedical studies, offering the unique capability to simultaneously emit both visible (VIS) and near-infrared (NIR) photons under continuous-wave (CW) NIR excitation. However, deep-tissue high-resolution imaging remains challenging due trade-off between VIS emission (higher resolution, limited penetration) (deeper penetration, lower resolution). Here we present a CW microscopy based on homologous dual-emission adaptive optics, leveraging Tm3+/Yb3+ co-doped UCNPs' dual 455 nm/800 nm emission: 800 for aberration measurement (guide-star) deep tissues at matching depths. Using home-built laser scanning microscope with 975 laser, achieved near-diffraction-limited (480 laterally) 500 μm depth mouse brain environment significant optical aberrations. This strategy expands applications innovates exploration of features.

Language: Английский

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Sidelobe-free deterministic 3D nanoscopy with λ/33 axial resolution

Light Science & Applications, Journal Year: 2025, Volume and Issue: 14(1)

Published: April 21, 2025

Abstract Deterministic three-dimensional (3D) super-resolution microscopy can achieve light-matter interaction in a small volume, but usually with the axial extension distinctly more elongated than lateral one. The isoSTED method combining two opposing objectives and multiple laser beams offer high at λ/12 level, cost of optical system complexity inherent sidelobes. high-order nonlinear effect by multiphoton excitation would benefit to sub-diffraction resolution as well suppress Herein, an easy-to-use, sidelobe-free deterministic 3D nanoscopy resolution, we developed purely physical strategy (UNEx-4Pi) fusion ultrahighly (UNEx) photon avalanching nanoparticles mirror-based bifocal vector field modulation (4Pi). theoretical studies UNEx-4Pi concept showed that main peak fluorescence spot became sharper its large sidelobe height was suppressed increasing nonlinearity. In addition, simplicity robustness were demonstrated utilizing mirror-assisted single-objective self-interference strategy. Experimentally, realized extremely constringent focal without sidelobes observed, achieving up λ/33 (26 nm) using one low-power CW beam. We also ability scheme bioimaging nuclear envelope BSC-1 cells stained imaged 32 nm. proposed will pave way for highly confined space, thereby advancing cutting-edge technologies like sensing, imaging, lithography, data storage.

Language: Английский

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MINFLUX nanoscopy enhanced with high-order vortex beams

Light Science & Applications, Journal Year: 2025, Volume and Issue: 14(1)

Published: May 6, 2025

Abstract Minimal photon fluxes (MINFLUX) nanoscopy has emerged as a transformative advancement in superresolution imaging, enabling unprecedented nanoscale observations across diverse biological scenarios. In this work, we propose, for the first time, that employing high-order vortex beams can significantly enhance performance of MINFLUX, surpassing limitations conventional MINFLUX using first-order beam. Our theoretical analysis indicates that, standard improve maximum localization precision by factor corresponding to their order, which approach sub-nanometer scale under optimal conditions, and raster scan allow wider field view while maintaining enhanced precision. These findings underscore potential elevate paving way towards ultra-high resolution imaging broad range applications.

Language: Английский

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Ensemble deep learning-enabled single-shot composite structured illumination microscopy (eDL-cSIM)

PhotoniX, Journal Year: 2025, Volume and Issue: 6(1)

Published: May 7, 2025

Abstract Structured illumination microscopy (SIM) has emerged as a powerful super-resolution technique for studying protein dynamics in live cells thanks to its wide-field imaging mode and high photon efficiency. However, conventional SIM requires at least nine raw images achieve reconstruction, which limits speed increases susceptibility rapid sample dynamics. Moreover, the reliance of on parameters algorithmic post-processing renders it vulnerable reconstruction artifacts, especially low signal-to-noise ratios. In this work, we propose single-shot composite structured method using ensemble deep learning (eDL-cSIM). Without modifying original setup, eDL-cSIM employs only one pattern generated by 6-beam interferometry. The resultant composite-coded image, contains multiplexed high-frequency spectral information beyond diffraction limit, is further processed predict high-quality, artifact-free super-resolved image. Experimental results demonstrate that integrates advantages various state-of-the-art neural networks, enabling robust image predictions across different specimen types or structures interest, outperforms classical physics-driven methods terms speed, quality environmental robustness, while avoiding intricate specialized procedures. These collective make promising tool fast live-cell with significantly reduced phototoxicity photobleaching.

Language: Английский

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High-performance Gd3Al4GaO12:Cr3+ phosphors for next-generation far-red LEDs

Materials Research Bulletin, Journal Year: 2023, Volume and Issue: 163, P. 112234 - 112234

Published: March 15, 2023

Language: Английский

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ISM-FLUX: MINFLUX with an array detector

Physical Review Research, Journal Year: 2023, Volume and Issue: 5(2)

Published: April 17, 2023

Single-molecule localization based on the concept of MINFLUX allows for molecular resolution imaging and tracking with a laser-scanning microscopy architecture. In MINFLUX, doughnut-shaped structured illumination beam sequentially excites sample, thereby minimizing uncertainty given number photons. However, has limited field view (FOV) therefore requires precise prelocalization step. We mitigate this problem by introducing single-molecule image scanning (smISM) concept. smISM, point detector microscope is replaced small single-photon array, leveraging benefits both detection. show via simulations that combining smISM (i.e., ISM-FLUX), we obtained between 1 15 nm over FOV more than 800 100 addition, multiple molecules simultaneously. calculate effect different parameters, such as relative position doughnut beams, pixels, photons, signal-to-background ratio, uncertainty. predict combination good precision enhanced robustness ISM-FLUX will help wide adoption MINFLUX.

Language: Английский

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