The emerging landscape of spatial profiling technologies

Nature Reviews Genetics, Год журнала: 2022, Номер 23(12), С. 741 - 759

Опубликована: Июль 20, 2022

Язык: Английский

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Whole-body cellular mapping in mouse using standard IgG antibodies

Nature Biotechnology, Год журнала: 2023, Номер 42(4), С. 617 - 627

Опубликована: Июль 10, 2023

Abstract Whole-body imaging techniques play a vital role in exploring the interplay of physiological systems maintaining health and driving disease. We introduce wildDISCO, new approach for whole-body immunolabeling, optical clearing mice, circumventing need transgenic reporter animals or nanobody labeling so overcoming existing technical limitations. identified heptakis(2,6-di- O -methyl)-β-cyclodextrin as potent enhancer cholesterol extraction membrane permeabilization, enabling deep, homogeneous penetration standard antibodies without aggregation. WildDISCO facilitates peripheral nervous systems, lymphatic vessels immune cells whole mice at cellular resolution by diverse endogenous proteins. Additionally, we examined rare proliferating effects biological perturbations, demonstrated germ-free mice. applied wildDISCO to map tertiary lymphoid structures context breast cancer, considering both primary tumor metastases throughout mouse body. An atlas high-resolution images showcasing nervous, vascular is accessible http://discotechnologies.org/wildDISCO/atlas/index.php .

Язык: Английский

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Light-sheets and smart microscopy, an exciting future is dawning

Communications Biology, Год журнала: 2023, Номер 6(1)

Опубликована: Май 9, 2023

Abstract Light-sheet fluorescence microscopy has transformed our ability to visualize and quantitatively measure biological processes rapidly over long time periods. In this review, we discuss current future developments in light-sheet that expect further expand its capabilities. This includes smart adaptive imaging schemes overcome traditional trade-offs, i.e., spatiotemporal resolution, field of view sample health. microscopy, a microscope will autonomously decide where, when, what how image. We assess image restoration techniques provide avenues these tradeoffs “open top” microscopes may enable multi-modal with high throughput. As such, predict fulfill an important role biomedical clinical the future.

Язык: Английский

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Whole-mouse clearing and imaging at the cellular level with vDISCO

Nature Protocols, Год журнала: 2023, Номер 18(4), С. 1197 - 1242

Опубликована: Янв. 25, 2023

Язык: Английский

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Spatial proteomics in three-dimensional intact specimens

Cell, Год журнала: 2022, Номер 185(26), С. 5040 - 5058.e19

Опубликована: Дек. 1, 2022

Язык: Английский

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EZ Clear for simple, rapid, and robust mouse whole organ clearing

eLife, Год журнала: 2022, Номер 11

Опубликована: Окт. 11, 2022

Tissue clearing for whole organ cell profiling has revolutionized biology and imaging exploration of organs in three-dimensional space without compromising tissue architecture. But complicated, laborious procedures, or expensive equipment, as well the use hazardous, organic solvents prevent widespread adoption these methods. Here, we report a simple rapid method, EZ Clear, that can clear adult mouse 48 hr just three steps. Samples stay at room temperature remain hydrated throughout process, preserving endogenous synthetic fluorescence, altering sample size. After wholemount imaging, samples processed with Clear be subjected to downstream applications, such embedding cryosectioning followed by standard histology immunofluorescent staining loss fluorescence signal from reporters. Furthermore, demonstrate brains successfully immunolabeled fluorescent while still retaining Overall, simplicity, speed, flexibility make it easy adapt implement diverse modalities biomedical research.

Язык: Английский

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Imagining the future of optical microscopy: everything, everywhere, all at once

Communications Biology, Год журнала: 2023, Номер 6(1)

Опубликована: Окт. 28, 2023

The optical microscope has revolutionized biology since at least the 17

Язык: Английский

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descSPIM: an affordable and easy-to-build light-sheet microscope optimized for tissue clearing techniques

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Июнь 12, 2024

Despite widespread adoption of tissue clearing techniques in recent years, poor access to suitable light-sheet fluorescence microscopes remains a major obstacle for biomedical end-users. Here, we present descSPIM (desktop-equipped SPIM cleared specimens), low-cost ($20,000-50,000), low-expertise (one-day installation by non-expert), yet practical do-it-yourself microscope as solution this bottleneck. Even the most fundamental configuration enables multi-color imaging whole mouse brains and cancer cell line-derived xenograft tumor mass visualization neurocircuitry, assessment drug distribution, pathological examination false-colored hematoxylin eosin staining three-dimensional manner. Academically open-sourced ( https://github.com/dbsb-juntendo/descSPIM ), allows routine samples minutes. Thus, dissemination will accelerate discoveries driven technologies.

Язык: Английский

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Three‐dimensional optically cleared tissue imaging for analyzing endoscopic images of gastrointestinal neoplasms (with video)

Digestive Endoscopy, Год журнала: 2025, Номер unknown

Опубликована: Фев. 3, 2025

Objectives To develop a procedure that matches magnifying endoscopic images with narrow‐band imaging to 3D tissue structures using tissue‐clearing technique and qualitatively quantitatively analyze specified in gastrointestinal neoplasms. Methods Endoscopically resected formalin‐fixed paraffin‐embedded tissues (three esophagus, four stomach, seven colon) were made transparent by ethyl cinnamate. They then subjected fluorescent staining of nuclei blood vessels followed confocal laser scanning microscope. A one‐to‐one correspondence between reconstructed was established crypts characteristic shapes as guides, the depth caliber measured. Results All optically cleared, which allowed visualization vascular all layers. In intraepithelial papillary capillary loops subepithelial networks identified. upper part polygonal surrounding pits observed, while lower part, surface epithelium ridge‐like coiled observed. honeycomb pit structure identified colon. Quantitative analysis showed various contrasts single continuous vessel image due different depths at tortuously ran. Conclusion We allow measure endoscopically visualized interest. This method is expected improve diagnosis further development technologies.

Язык: Английский

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Nanoscale Resolution Imaging of Whole Mouse Embryos Using Expansion Microscopy

ACS Nano, Год журнала: 2025, Номер unknown

Опубликована: Фев. 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.

Язык: Английский

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