Application of Light-Sheet Mesoscopy to Image Host-Pathogen Interactions in Intact Organs DOI Creative Commons
Eliana Battistella, Juan F. Quintana, Gail McConnell

et al.

Frontiers in Cellular and Infection Microbiology, Journal Year: 2022, Volume and Issue: 12

Published: June 14, 2022

Human African Trypanosomiasis (HAT) is a disease caused by the extracellular parasite Trypanosoma brucei that affects central nervous system (CNS) during chronic stage of infection, inducing neuroinflammation, coma, and death if left untreated. However, little known about structural change happening in brain as result infection. So far, infection-induced neuroinflammation has been observed with conventional methods, such immunohistochemistry, electron microscopy, 2-photon microscopy only small portions brain, which may not be representative disease. In this paper, we have used newly-developed light-sheet illuminator to image level chronically infected mice compared it naïve controls. This was developed for imaging combination Mesolens objective lens, providing fast sub-cellular resolution tens mm 3 -large volumes. The mouse specimens were cleared using CUBIC+, followed antibody staining locate Glial Fibrillary Acid Protein (GFAP) expressing cells, primarily astrocytes ependymocytes, here proxy cell reactivity gliosis. large capture volume allowed us detect GFAP + cells spatially resolve response T. Based on morphometric analyses spatial distribution our data demonstrates significant increase dendrite branching around lateral ventricle, well dorsal ventral third ventricles, are negatively correlated branch extension distal sites from circumventricular spaces. To knowledge, first report highlighting potential mesoscopy characterise inflammatory responses parasitic infection at cellular intact organs, opening new avenues development mesoscale techniques study host-pathogen interactions.

Language: Английский

Multi‐Photon Microscopy DOI

Jeremy Sanderson

Current Protocols, Journal Year: 2023, Volume and Issue: 3(1)

Published: Jan. 1, 2023

In this series of papers on light microscopy imaging, we have covered the fundamentals microscopy, super-resolution and lightsheet microscopy. This last review covers multi-photon with a brief reference to intravital imaging Brainbow labeling. Multi-photon is often referred as two-photon Indeed, using by far most common way thick tissues; however, it theoretically possible use higher number photons, three-photon possible. Therefore, titled "multi-photon microscopy." Another term for describing "non-linear" because fluorescence intensity at focal spot depends upon average squared rather than intensity; hence, non-linear optics (NLO) an alternative name It relationship (or third exponential power in case excitation) that determines axial optical sectioning capability imaging. paper, necessity or explained, method described. Advice also given what fluorescent markers other practical aspects tissues. The technique discussed. concludes description mouse. © 2023 Wiley Periodicals LLC.

Language: Английский

Citations

16
In vivo three-dimensional brain imaging with chemiluminescence probes in Alzheimer’s disease models DOI Creative Commons
Jing Zhang, Carly Wickizer, Weihua Ding

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2023, Volume and Issue: 120(50)

Published: Dec. 4, 2023

Optical three-dimensional (3D) molecular imaging is highly desirable for providing precise distribution of the target-of-interest in disease models. However, such 3D still far from wide applications biomedical research; brain optical imaging, particular, has rarely been reported. In this report, we designed chemiluminescence probes with high quantum yields, relatively long emission wavelengths, and signal-to-noise ratios to fulfill requirements vivo. With assistance density-function theory (DFT) computation, ADLumin-Xs by locking up rotation double bond via fusing furan ring phenyl ring. Our results showed that ADLumin-5 had a yield could bind amyloid beta (Aβ). Remarkably, ADLumin-5's radiance intensity areas reach 4 × 10

Language: Английский

Citations

15
Challenges and advances in optical 3D mesoscale imaging DOI Creative Commons
Sebastian Munck, Christopher Cawthorne, Abril Escamilla‐Ayala

et al.

Journal of Microscopy, Journal Year: 2022, Volume and Issue: 286(3), P. 201 - 219

Published: May 5, 2022

Abstract Optical mesoscale imaging is a rapidly developing field that allows the visualisation of larger samples than possible with standard light microscopy, and fills gap between cell organism resolution. It spans from advanced fluorescence micrometric clusters to centimetre‐size complete organisms. However, volume specimens, new problems arise. Imaging deeper into tissues at high resolution poses challenges ranging optical distortions shadowing opaque structures. This manuscript discusses latest developments in highlights limitations, namely labelling, clearing, absorption, scattering, also sample handling. We then focus on approaches seek turn more quantitative technique, analogous tomography medical imaging, highlighting future role for digital physical phantoms as well artificial intelligence.

Language: Английский

Citations

18
Multiscale Three‐Dimensional Evaluation and Analysis of Murine Lung Vasculature From Macro‐ to Micro‐Structural Level DOI Creative Commons
Birger Tielemans,

Nora F. Marain,

Axelle Kerstens

et al.

Pulmonary Circulation, Journal Year: 2025, Volume and Issue: 15(1)

Published: Jan. 1, 2025

ABSTRACT The pulmonary vasculature plays a pivotal role in the development and progress of chronic lung diseases. Due to limitations conventional two‐dimensional histological methods, complexity detailed anatomy blood circulation might be overlooked. In this study, we demonstrate practical use optical serial block face imaging (SBFI), ex vivo microcomputed tomography (micro‐CT), nondestructive for visualization quantification circulation's 3D architecture from macro‐ micro‐structural levels murine samples. We that SBFI can provide rapid, cost‐effective, label‐free mouse macrostructures large vessels. Micro‐CT offers high‐resolution captures microvascular (pre)capillary structures, with microstructural quantification. Optical microscopy techniques such as projection (OPT) light sheet fluorescence microscopy, allows noninvasive, mesoscopic optically cleared lungs, still enabling microscopic reconstruction down precapillary level. By integrating SBFI, micro‐CT, framework understanding circulation, emphasis on vascular pruning rarefaction. Our study showcases applicability complementarity these organ‐level imaging, offering researchers flexibility selecting optimal approach based their specific requirements. conclusion, propose 3D‐directed approaches whole‐organ view health disease, advance our current knowledge diseases affecting vasculature.

Language: Английский

Citations

0
Distributed and networked analysis of volumetric image data for remote collaboration of microscopy image analysis DOI
Alain Chen, Shuo Han, Soonam Lee

et al.

Journal of Medical Imaging, Journal Year: 2025, Volume and Issue: 12(02)

Published: March 11, 2025

The advancement of high-content optical microscopy has enabled the acquisition very large three-dimensional (3D) image datasets. analysis these volumes requires more computational resources than a biologist may have access to in typical desktop or laptop computers. This is especially true if machine learning tools are being used for analysis. With increased amount data and complexity, there need accessible, easy-to-use, efficient network-based 3D processing system. distributed networked volumetric (DINAVID) system was developed enable remote images biologists. We present an overview DINAVID compare it other currently available designed using open-source two main sub-systems, visualization with simple web interface that allows biologists upload visualization. enables model center hosting users analyzing those volumes, without manage any resources. system, tools, analyze visualize remotely also provides several including pre-processing segmentation models.

Language: Английский

Citations

0
暗场光散射成像-表面增强拉曼散射联用技术及其分析检测应用 DOI

兰新宇 Lan Xinyu,

翁国军 Weng Guojun,

李新 Li Xin

et al.

Laser & Optoelectronics Progress, Journal Year: 2025, Volume and Issue: 62(8), P. 0800003 - 0800003

Published: Jan. 1, 2025

Citations

0
High Content Imaging Applications of Advanced and Translational Disease Models in Drug Discovery and Development DOI
Weiyingqi Cui,

Mariam Haffa,

Francesco Massai

et al.

Royal Society of Chemistry eBooks, Journal Year: 2025, Volume and Issue: unknown, P. 168 - 208

Published: April 30, 2025

There has been increasing interest in disease models with enhanced physiological fidelity. This led to the development of new methods for generating advanced utilizing primary cells and renewable sources, such as induced pluripotent stem organoids. Furthermore, combining these types high content imaging is expected positively impact all stages drug discovery pipeline. Since data rich assays can uncover nuanced cellular response perturbation. In this review, we focus on recent application models, covering general considerations cell source, culture format screening, preclinical studies translational applications, functional precision medicine approaches.

Language: Английский

Citations

0
Quantitative characterization of tumor cell traction force on extracellular matrix by hydrogel microsphere stress sensor DOI

Xingquan Ma,

Cong Wang, Changchun Ji

et al.

Biotechnology and Bioengineering, Journal Year: 2024, Volume and Issue: 121(6), P. 1820 - 1830

Published: Feb. 26, 2024

Cell traction force (CTF) is a kind of active that cell senses external environment and actively applies to the contact matrix which currently representative stress in cell-extracellular (ECM) interaction. Studying distribution variation CTF during cell-ECM interaction help explain impact physical factors on behaviors from perspective mechanobiology. However, most strategies characterizing are still limited by measurement needs three-dimensional (3D), quantitative characteristics vivo condition. Microsphere sensor (MSS) as new type technology capable realizing characterization 3D vivo. Herein, we employed microfluidic platform design fabricate MSS possesses adjustable fluorescent performances, properties, size ranges for better applicable different cells (3T3, A549). Focusing common tumor (adhesion, spreading, migration) process metastasis, chose SH-SY5Y research object this work. We calculated with profile demonstrate normal shear can determined behaviors. Additionally, also regulate adhesion, migration states. Based method, CFT health disease be achieved, further study explore potential mechanism

Language: Английский

Citations

3
Three-Dimensional Optical Imaging of Internal Deformations in Polymeric Microscale Mechanical Metamaterials DOI Creative Commons
Brian W. Blankenship, Timon Meier,

Naichen Zhao

et al.

Nano Letters, Journal Year: 2024, Volume and Issue: 24(9), P. 2735 - 2742

Published: Jan. 26, 2024

Recent advances in two-photon polymerization fabrication processes are paving the way to creating macroscopic metamaterials with microscale architectures, which exhibit mechanical properties superior their bulk material counterparts. These typically feature lightweight, complex patterns such as lattice or minimal surface structures. Conventional tools for investigating these structures, scanning electron microscopy, cannot easily probe internal features of critical a comprehensive assessment behavior. In turn, we demonstrate an optical confocal microscopy-based approach that allows high-resolution imaging deformations and fracture under load. We validate this technique by exemplary metamaterial structure 80 × μm3 size. This can be extended other systems holds significant promise enhance our understanding real-world performance loading conditions.

Language: Английский

Citations

2
Multimodal optical mesoscopy reveals the quantity and spatial distribution of Gram‐positive biofilms in ex vivo tonsils DOI Creative Commons
Megan Clapperton,

Tash Kunanandam,

Catalina D. Florea

et al.

Journal of Microscopy, Journal Year: 2024, Volume and Issue: 295(2), P. 121 - 130

Published: Jan. 31, 2024

Abstract Biofilms are known to be present in tonsils, but little is about their spatial location and size distribution throughout the tonsil. Studies of biofilms tonsil specimens have thus far been limited either high‐magnification methods such as electron microscopy, which enables high‐resolution imaging only from a tiny tissue volume, or lower magnification techniques light allow larger with poor resolution. To overcome these limitations, we report use multimodal optical mesoscopy visualise quantify number Gram‐positive fresh, excised paediatric tonsils. This methodology supports simultaneous both host whole mounts up 5 mm × 3 subcellular resolution throughout. A quantitative assessment 36 revealed no statistically significant difference between biofilm presence on surface interior new mesoscale approach may prove useful understanding role tonsillar diseases other infections.

Language: Английский

Citations

2