Nanoscale Resolution Imaging of Whole Mouse Embryos Using Expansion Microscopy

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

Published: Feb. 18, 2025

Nanoscale imaging of whole vertebrates is essential for the systematic understanding human diseases, yet this goal has not been achieved. Expansion microscopy (ExM) an attractive option accomplishing aim; however, expansion even mouse embryos at mid- and late-developmental stages, which have fewer calcified body parts than adult mice, to be demonstrated due challenges expanding tissues. Here, we introduce a state-of-the-art ExM technique, termed whole-body ExM, that utilizes cyclic digestion. This technique allows super-resolution, volumetric anatomical structures, proteins, endogenous fluorescent proteins (FPs) within embryonic neonatal mice by them 4-fold. The key feature alternating application two enzyme compositions repeated multiple times. Through simple repetition digestion process with increasing number cycles, various stages up E18.5, display dramatic difference in content tissues compared embryos, are expanded without further laborious optimization. Furthermore, ExM's ability retain FP signals visualization neuronal structures transgenic mice. Whole-body could facilitate studies molecular changes vertebrates.

Language: Английский

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Tissue clearing and 3D reconstruction of digitized, serially sectioned slides provide novel insights into pancreatic cancer

Med, Journal Year: 2023, Volume and Issue: 4(2), P. 75 - 91

Published: Jan. 24, 2023

Language: Английский

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Principles of deep immunohistochemistry for 3D histology

Cell Reports Methods, Journal Year: 2023, Volume and Issue: 3(5), P. 100458 - 100458

Published: April 20, 2023

Deep immunohistochemistry (IHC) is a nascent field in three-dimensional (3D) histology that seeks to achieve thorough, homogeneous, and specific staining of intact tissues for visualization microscopic architectures molecular compositions at large spatial scales. Despite the tremendous potential deep IHC revealing molecule-structure-function relationships biology establishing diagnostic prognostic features pathological samples clinical practice, complexities variations methodologies may hinder its use by interested users. We provide unified framework immunostaining techniques discussing theoretical considerations physicochemical processes involved, summarizing principles applied contemporary methods, advocating standardized benchmarking scheme, highlighting unaddressed issues future directions. By providing essential information guide investigators customizing immunolabeling pipelines, we also seek facilitate adoption researchers address wide range research questions.

Language: Английский

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Visualization of metabolites and microbes at high spatial resolution using MALDI mass spectrometry imaging and in situ fluorescence labeling

Nature Protocols, Journal Year: 2023, Volume and Issue: 18(10), P. 3050 - 3079

Published: Sept. 6, 2023

Language: Английский

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Reflective multi-immersion microscope objectives inspired by the Schmidt telescope

Nature Biotechnology, Journal Year: 2023, Volume and Issue: 42(1), P. 65 - 71

Published: March 30, 2023

Abstract Imaging large, cleared samples requires microscope objectives that combine a large field of view (FOV) with long working distance (WD) and high numerical aperture (NA). Ideally, such should be compatible wide range immersion media, which is challenging to achieve conventional lens-based objective designs. Here we introduce the multi-immersion ‘Schmidt objective’ consisting spherical mirror an aspherical correction plate as solution this problem. We demonstrate multi-photon variant Schmidt all homogeneous media achieves NA 1.08 at refractive index 1.56, 1.1-mm FOV 11-mm WD. highlight its versatility by imaging in various ranging from air water benzyl alcohol/benzyl benzoate, dibenzyl ether ethyl cinnamate neuronal activity larval zebrafish vivo. In principle, concept can extended any modality, including wide-field, confocal light-sheet microscopy.

Language: Английский

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A biologist’s guide to planning and performing quantitative bioimaging experiments

PLoS Biology, Journal Year: 2023, Volume and Issue: 21(6), P. e3002167 - e3002167

Published: June 27, 2023

Technological advancements in biology and microscopy have empowered a transition from bioimaging as an observational method to quantitative one. However, biologists are adopting these experiments become more complex, researchers need additional expertise carry out this work rigorous reproducible manner. This Essay provides navigational guide for experimental aid understanding of sample preparation through image acquisition, analysis, data interpretation. We discuss the interconnectedness steps, each, we provide general recommendations, key questions consider, links high-quality open-access resources further learning. synthesis information will empower plan execute efficiently.

Language: Английский

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Neuroscience in peripheral cancers: tumors hijacking nerves and neuroimmune crosstalk

MedComm, Journal Year: 2024, Volume and Issue: 5(11)

Published: Oct. 31, 2024

Abstract Cancer neuroscience is an emerging field that investigates the intricate relationship between nervous system and cancer, gaining increasing recognition for its importance. The central governs development of directly affects brain tumors, peripheral (PNS) shapes tumor microenvironment (TME) tumors. Both systems are crucial in cancer initiation progression, with recent studies revealing a more role PNS within TME. Tumors not only invade nerves but also persuade them through remodeling to further promote malignancy, creating bidirectional interaction cancers. Notably, immune cells contribute this communication, forming triangular influences protumor inflammation effectiveness immunotherapy. This review delves into mechanisms connecting focusing on how various cell types influence nerve‒tumor interactions, emphasizing clinical relevance nerve‒immune dynamics. By deepening our understanding interplay nerves, cells, has potential reshape biology insights, inspire innovative therapies, improve outcomes patients.

Language: Английский

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Obesity-Related Neuroinflammation: Magnetic Resonance and Microscopy Imaging of the Brain

International Journal of Molecular Sciences, Journal Year: 2022, Volume and Issue: 23(15), P. 8790 - 8790

Published: Aug. 8, 2022

Obesity is a major risk factor of Alzheimer's disease and related dementias. The principal feature dementia loss neurons brain atrophy. mechanistic links between obesity the neurodegenerative processes dementias are not fully understood, but recent research suggests that obesity-related systemic inflammation subsequent neuroinflammation may be involved. Adipose tissues release multiple proinflammatory molecules (fatty acids cytokines) impact blood vessel cells, inducing low-grade can transition to tissues, including brain. Inflammation in brain-neuroinflammation-is one key elements pathobiology disorders; it characterized by activation microglia, resident immune cells brain, structural functional changes other forming parenchyma, neurons. Such cellular have been shown animal models with direct methods, such as confocal microscopy. In humans, less tangible, only indirect methods magnetic resonance (MR) imaging usually used. these studies, associated lower volumes cerebral gray matter, cortex, hippocampus, well altered tissue MR properties (suggesting microstructural variations molecular composition). How human observed using relate seen microscopy understood. This review describes current understanding context obesity-induced inflammation, highlights need for bridge studies.

Language: Английский

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Nondestructive 3D Pathology with Light-Sheet Fluorescence Microscopy for Translational Research and Clinical Assays

Annual Review of Analytical Chemistry, Journal Year: 2023, Volume and Issue: 16(1), P. 231 - 252

Published: Feb. 28, 2023

In recent years, there has been a revived appreciation for the importance of spatial context and morphological phenotypes both understanding disease progression guiding treatment decisions. Compared with conventional 2D histopathology, which is current gold standard medical diagnostics, nondestructive 3D pathology offers researchers clinicians ability to visualize orders magnitude more tissue within their natural volumetric context. This enabled by rapid advances in tissue-preparation methods, high-throughput microscopy instrumentation, computational tools processing these massive feature-rich data sets. Here, we provide brief overview many technical along remaining challenges be overcome. We also speculate on future as applied translational investigations, preclinical drug development, clinical decision-support assays.

Language: Английский

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Preserving extracellular space for high-quality optical and ultrastructural studies of whole mammalian brains

Cell Reports Methods, Journal Year: 2023, Volume and Issue: 3(7), P. 100520 - 100520

Published: July 1, 2023

Analysis of brain structure, connectivity, and molecular diversity relies on effective tissue fixation. Conventional fixation causes extracellular space (ECS) loss, complicating the segmentation cellular objects from electron microscopy datasets. Previous techniques for preserving ECS in mammalian brains utilizing high-pressure perfusion can give inconsistent results owing to variations hydrostatic pressure within vasculature. A more reliable protocol that uniformly preserves throughout whole would greatly benefit a wide range neuroscience studies. Here, we report straightforward transcardial strategy rodent brain. No special setup is needed besides sequential solution changes, offers excellent reproducibility. In addition better capturing ultrastructure, preservation has many downstream advantages such as accelerating heavy-metal staining microscopy, improving detergent-free immunohistochemistry correlated light facilitating lipid removal clearing.

Language: Английский

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