A Random Angle Error Interference Eliminating Method for Grating Interferometry Measurement Based on Symmetry Littrow Structure

Laser & Photonics Review, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 28, 2025

Abstract All linear motion will have random angle errors, which lead to displacement detection result deviating from the true value. To solve this common problem in field of optical precision measurement, study proposes a symmetry Littrow grating interferometry measurement method (SLGIM). SLGIM uses high‐precision planar diffraction gratings and place them on both sides central main system build two complementary arms with equal paths. There is mode suppression relationship between arms, so can eliminate error interference improve detection. The experiment results show that when same applied, compared traditional system, mean value decreases 25.8%, standard deviation 37.5%, range 33.7%. after using structure proposed achieves nanometer‐level precision.

Language: Английский

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A highly stable monomeric red fluorescent protein for advanced microscopy

Nature Methods, Journal Year: 2025, Volume and Issue: unknown

Published: April 17, 2025

Language: Английский

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Multi-resolution analysis enables fidelity-ensured deconvolution for fluorescence microscopy

eLight, Journal Year: 2024, Volume and Issue: 4(1)

Published: Aug. 6, 2024

Abstract Fluorescence microscopic imaging is essentially a convolution process distorted by random noise, limiting critical parameters such as speed, duration, and resolution. Though algorithmic compensation has shown great potential to enhance these pivotal aspects, its fidelity remains questioned. Here we develop physics-rooted computational resolution extension denoising method with ensured fidelity. Our approach employs multi-resolution analysis (MRA) framework extract the two main characteristics of fluorescence images against noise: across-edge contrast, along-edge continuity. By constraining features in model-solution using framelet curvelet, MRA deconvolution algorithms, which improve signal-to-noise ratio (SNR) up 10 dB higher than spatial derivative based penalties, can provide two-fold fidelity-ensured improvement rather artifact-prone Richardson-Lucy inference. We demonstrate our methods performance various diffraction-limited super-resolution microscopies fidelity, enabling accomplishments more challenging tasks.

Language: Английский

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Expanding super-resolution imaging versatility in organisms with multi-confocal image scanning microscopy

National Science Review, Journal Year: 2024, Volume and Issue: 11(9)

Published: Aug. 13, 2024

Resolving complex three-dimensional (3D) subcellular dynamics noninvasively in live tissues demands imaging tools that balance spatiotemporal resolution, field-of-view and phototoxicity. Image scanning microscopy (ISM), as an advancement of confocal laser microscopy, provides a 2-fold 3D resolution enhancement. Nevertheless, the relatively low speed has been major obstacle for ISM to be further employed vivo biological tissues. Our proposed solution, multi-confocal image (MC-ISM), aims overcome limitations existing techniques terms balancing by optimizing pinhole diameter pitch, eliminating out-of-focus signals, introducing frame reduction reconstruction algorithm. The is increased 16 times compared with multifocal structured illumination microscopy. We propose single-galvo scan, akin Archimedes spiral spinning disk systems, ensure high-speed high-accuracy scan without galvanometer's inertial motion. Benefitting from its high photon efficiency, MC-ISM allows continuous mitochondria cells 1000 frames apparent phototoxicity, reaching depth 175 μm. Noteworthy, enables observation inner membrane structure living Arabidopsis hypocotyl first time, demonstrating outstanding performance.

Language: Английский

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Fast reconstruction and optical-sectioning three-dimensional structure illumination microscopy

The Innovation, Journal Year: 2025, Volume and Issue: unknown, P. 100757 - 100757

Published: Jan. 1, 2025

Public summary•3D structured illumination microscopy is enhanced integrating spatial reconstruction and optical sectioning.•Reconstruction speed up by nearly three orders of magnitude fidelity has been greatly improved.•We achieve high-throughput near-real-time 3D super-resolution imaging with high fidelity.AbstractThree-dimensional structure (3DSIM) a popular method for observing subcellular/cellular structures or animal/plant tissues gentle phototoxicity super-resolution. However, its time-consuming process poses challenges real-time observation. Moreover, traditional 3DSIM typically requires more than six z layers successful susceptible to defocused backgrounds. This great gap between single-layer 2DSIM 6-layer 3DSIM, limits the observation thicker samples. To address these limitations, we developed FO-3DSIM, novel that integrates spatial-domain optical-sectioning SIM. FO-3DSIM enhances 855.7 times superior performance limited under It retains high-fidelity, low-photon capabilities our previously proposed Open-3DSIM. Utilizing fast sectioning, achieved large field-of-view (FOV) mouse kidney actin, covering region 0.453 mm × 2.75 μm within 23 min acquisition 13 reconstruction. Near was demonstrated in live actin FO-3DSIM. Our approach reduces photodamage through layer reconstruction, allowing ER tubes just layers. We anticipate will pave way near real-time, FOV 6D imaging, encompassing xyz super-resolution, multi-color, long-term, polarization less photodamage, removed backgrounds, reduced time.Graphical abstract

Language: Английский

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HBimmCue: A Versatile Fluorescent Probe for Multi‐Scale Imaging of Lipid Polarity and Membrane Order in Inner Mitochondrial Membrane

Advanced Science, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 9, 2025

Abstract Mitochondrial membrane environmental dynamics are crucial for understanding function, yet high‐resolution observation remains challenging. Here, HBimmCue is introduced as a fluorescent probe localized to inner mitochondrial (IMM) that reports lipid polarity and order changes, which correlate with cellular respiration levels. Using fluorescence lifetime imaging microscopy (FLIM), IMM heterogeneity uncovered across scales, from nanoscale structures within individual mitochondria mouse pre‐implantation embryos. At the sub‐organelle level, stimulated emission depletion (STED)‐FLIM highlights variations IMM. sub‐cellular reduced observed in damaged marked lysosomal degradation distinct distributions identified neurons disease models. Additionally, metabolic dysfunction associated oocytes aging reprogramming zygote blastocyst detected. Together, work demonstrates broad applicability of HBimmCue, offering new paradigm investigating level at multiple scales.

Language: Английский

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How are mitochondrial nucleoids trafficked?

Trends in Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

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Dual-Modality Imaging Unveil Inner Mitochondrial Membrane Viscosity and Respiratory Dynamics in Mitophagy

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 27, 2025

Mitophagy is a vital lysosome-dependent process that maintains mitochondrial integrity and cellular homeostasis, where respiration inner membrane (IMM) viscosity play key roles. Despite its critical importance, achieving high-resolution dynamic visualization of IMM during mitophagy remains significant challenge. In this study, we designed two innovative fluorescent probes: SiR-C8, viscosity-sensitive rotor-type probe based on silicon-rhodamine, specifically targeting the IMM, OR-ATP, rhodamine-derived utilizing an intramolecular spirolactam structure to respond ATP levels. Leveraging fluorescence intensity lifetime dual-modality imaging, successfully enabled high-resolution, real-time monitoring mitophagy. Remarkably, our results unveiled progressive increase in alongside attenuation induced by starvation, carbonyl cyanide, m-chlorophenyl hydrazone (CCCP), Oligomycin. Significantly, structured illumination microscopy super-resolution have uncovered novel quality control mechanism which lysosomes selectively engulf locally damaged regions. This discovery provides insights into intricate processes governing introduces platform for studying dynamics, dysfunction, their implications homeostasis pathology.

Language: Английский

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Molecular machineries shaping the mitochondrial inner membrane

Nature Reviews Molecular Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: May 14, 2025

Language: Английский

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Harnessing an MMP-Independent NIR Probe Unveiling the Different Mitochondrial Cristae Changes during Mitophagy and Ferroptosis under STED Microscopy

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 2, 2025

Mitochondrial cristae remain dynamic structures in order to adapt various physiopathologic processes (e.g., mitophagy and ferroptosis); thus, visualizing tracking different changes of are crucial for a deeper understanding these processes. Fluorescent probes that can realize long-term visualization mitochondrial under stimulated emission depletion (STED) microscopy powerful tools their in-depth research. However, there few reports on such probes, constructions challenging. Here, we reported robust squaraine probe (CSN) the physiological pathological using STED microscopy. The lipophilic unit CSN enabled it firmly immobilize mitochondria via hydrophobic interaction, which let labeling ability independent membrane potential (MMP). Using CSN, were clearly observed at resolution 52 nm Furthermore, was successfully applied track destruction during autophagy ferroptosis. Interestingly, found mitophagy, first underwent swelling rupture, then partial vacuolization, finally complete whereas ferroptosis, gradual reduction number cristae, fracture, vacuolization. This work revealed difference provided insights into two We believed could serve as desirable tool intracellular activity

Language: Английский

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