In situ isotropic 3D imaging of vasculature perfusion specimens using x‐ray microscopic dual‐energy CT DOI Creative Commons
Stephan Handschuh, Ursula Reichart, Stefan Kummer

и другие.

Journal of Microscopy, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 6, 2024

Abstract Ex vivo x‐ray angiography provides high‐resolution, three‐dimensional information on vascular phenotypes down to the level of capillaries. Sample preparation for ex starts with removal blood from system, followed by perfusion an dense contrast agent mixed a carrier such as gelatine or polymer. Subsequently, micro‐architecture harvested organs is imaged in intact fixed organ. In present study, we novel microscopic dual‐energy CT (microDECT) imaging protocols that allow visualise and analyse microvasculature situ reference morphology hard soft tissue. We show spectral µAngiofil Micropaque barium sulphate perfused specimens allows effective separation vasculature mineralised skeletal tissues. Furthermore, demonstrate counterstaining using established agents depict vessels together Phosphotungstic acid (PTA) used counterstain shows excellent both sulphate–perfused specimens. A Sorensen‐buffered PTA protocol introduced specimens, polyurethane polymer susceptible artefacts when conventional staining solutions. Finally, counterstained samples can be automatically processed into three separate image channels (skeletal tissue, stained tissue), which offers multiple new options data analysis. The presented microDECT workflows are suited tools screen quantify implemented various correlative pipelines target regions interest downstream light investigation.

Язык: Английский

Frontiers in artificial intelligence‐directed light‐sheet microscopy for uncovering biological phenomena and multiorgan imaging DOI Creative Commons
Enbo Zhu,

Yan‐Ruide Li,

Samuel Margolis

и другие.

View, Год журнала: 2024, Номер 5(5)

Опубликована: Сен. 3, 2024

Light-sheet fluorescence microscopy (LSFM) introduces fast scanning of biological phenomena with deep photon penetration and minimal phototoxicity. This advancement represents a significant shift in 3-D imaging large-scale tissues 4-D (space + time) small live animals. The large data associated LSFM requires efficient acquisition analysis the use artificial intelligence (AI)/machine learning (ML) algorithms. To this end, AI/ML-directed is an emerging area for multi-organ tumor diagnostics. review will present development highlight various configurations designs multi-scale imaging. Optical clearance techniques be compared effective reduction light scattering optimal deep-tissue further depict diverse range research translational applications, from organisms to diagnosis. In addition, address reconstruction, including application convolutional neural networks (CNNs) generative adversarial (GANs). summary, advancements have enabled post-imaging reconstruction analyses, underscoring LSFM's contribution advancing fundamental research.

Язык: Английский

Процитировано

6
Scattering Reduction and Axial Resolution Enhancement in Light‐Sheet Fluorescence Microscopy DOI
Yang Zhang, Runze Li, Xianghua Yu

и другие.

Journal of Biophotonics, Год журнала: 2025, Номер unknown

Опубликована: Фев. 23, 2025

ABSTRACT Light‐sheet fluorescence microscopy (LSFM) provides an ideal tool for long‐term observation of live specimens due to its low photodamage and fast volumetric imaging speed. The wavefront distortions in the illumination path LSFM will reduce intensity broaden light‐sheet thickness, thereby degrading image quality. We propose use shaping technique scattering effect shrink thickness. Scanning refocused laser beam generate LS improves both axial resolution. resolution can be further enhanced by subtracting two images captured via double scanning samples with uncorrected scattered each slice. is improved from 2.2 ± 0.3 1.5 0.2 μm across field view 270 × μm. effectiveness subtraction method demonstrated fluorescent beads Aspergillus conidiophores behind a medium.

Язык: Английский

Процитировано

0
Near-Infrared Emissive π-Conjugated Oligomer Nanoparticles for Three- and Four-Photon Deep-Brain Microscopic Imaging Beyond 1700 nm Excitation DOI

Lijun Kan,

Yingxian Zhang, Yu Luo

и другие.

ACS Nano, Год журнала: 2024, Номер unknown

Опубликована: Сен. 19, 2024

High-resolution visualization of the deep brain is still a challenging and very significant issue. Multiphoton microscopy (MPM) holds great promise for high-spatiotemporal deep-tissue imaging under NIR-III NIR-IV excitation. However, thus far, their applications have been seriously restricted by scarcity efficient organic probes. Herein, we designed synthesized two donor-acceptor-donor-type conjugated small molecules (

Язык: Английский

Процитировано

1
Protocol for three-dimensional whole-mount imaging of the vascular network in the intestinal muscle DOI Creative Commons
Sandra Schrenk, Lindsay J. Bischoff, Elisa Boscolo

и другие.

STAR Protocols, Год журнала: 2024, Номер 5(3), С. 103170 - 103170

Опубликована: Июль 4, 2024

Three-dimensional (3D) imaging of vascular networks is essential for the investigation patterning and organization. Here, we present a step-by-step protocol 3D visualization vasculature within whole-mount preparations mouse intestinal muscularis propria layer. We then detail quantitative analysis resulting images parameters such as vessel density, diameter, number endothelial cells, proliferation. The can be easily extended to study cell-cell interactions neuro-vascular or immune-vascular interactions. For complete details on use execution this protocol, please refer Schrenk et al.

Язык: Английский

Процитировано

0
In situ isotropic 3D imaging of vasculature perfusion specimens using x‐ray microscopic dual‐energy CT DOI Creative Commons
Stephan Handschuh, Ursula Reichart, Stefan Kummer

и другие.

Journal of Microscopy, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 6, 2024

Abstract Ex vivo x‐ray angiography provides high‐resolution, three‐dimensional information on vascular phenotypes down to the level of capillaries. Sample preparation for ex starts with removal blood from system, followed by perfusion an dense contrast agent mixed a carrier such as gelatine or polymer. Subsequently, micro‐architecture harvested organs is imaged in intact fixed organ. In present study, we novel microscopic dual‐energy CT (microDECT) imaging protocols that allow visualise and analyse microvasculature situ reference morphology hard soft tissue. We show spectral µAngiofil Micropaque barium sulphate perfused specimens allows effective separation vasculature mineralised skeletal tissues. Furthermore, demonstrate counterstaining using established agents depict vessels together Phosphotungstic acid (PTA) used counterstain shows excellent both sulphate–perfused specimens. A Sorensen‐buffered PTA protocol introduced specimens, polyurethane polymer susceptible artefacts when conventional staining solutions. Finally, counterstained samples can be automatically processed into three separate image channels (skeletal tissue, stained tissue), which offers multiple new options data analysis. The presented microDECT workflows are suited tools screen quantify implemented various correlative pipelines target regions interest downstream light investigation.

Язык: Английский

Процитировано

0