Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain DOI Open Access
Ya‐Hui Lin, Liwen Wang, Yen‐Hui Chen

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Dec. 2, 2022

Abstract Tissue clearing and labeling techniques have revolutionized brain-wide imaging analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce HIF-Clear, a novel method for efficiently centimeter-thick FFPE specimens using elevated temperature concentrated detergents. HIF-Clear with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in whole mouse brain, is able be used as the evaluation of disease treatment efficiency. also supports expansion microscopy can performed on non-sectioned 15-year-old specimen, well 3-month brain. Thus, represents feasible approach researching archived future neuroscientific 3D neuropathological analyses.

Language: Английский

Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain DOI Open Access
Ya‐Hui Lin, Liwen Wang, Yen‐Hui Chen

et al.

Published: Jan. 15, 2024

Tissue clearing and labeling techniques have revolutionized brain-wide imaging analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce MOCAT, a novel method for efficiently centimeter-thick FFPE specimens using elevated temperature concentrated detergents. MOCAT with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in whole mouse brain, is able be used as the evaluation of disease treatment efficiency. also supports expansion microscopy can performed on non-sectioned 15-year-old specimen. Thus, represents feasible approach researching archived future neuroscientific 3D neuropathological analyses.

Language: Английский

Citations

0
Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain DOI Open Access
Ya‐Hui Lin, Liwen Wang, Yen‐Hui Chen

et al.

Published: March 8, 2024

Tissue clearing and labeling techniques have revolutionized brain-wide imaging analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce MOCAT, a novel method for efficiently centimeter-thick FFPE specimens using elevated temperature concentrated detergents. MOCAT with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in whole mouse brain, is able be used as the evaluation of disease treatment efficiency. also supports expansion microscopy can performed on non-sectioned 15-year-old specimen. Thus, represents feasible approach researching archived future neuroscientific 3D neuropathological analyses.

Language: Английский

Citations

0
Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain DOI Open Access
Ya‐Hui Lin, Liwen Wang, Yen‐Hui Chen

et al.

Published: April 4, 2024

Tissue clearing and labeling techniques have revolutionized brain-wide imaging analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce HIF-Clear, a novel method for efficiently centimeter-thick FFPE specimens using elevated temperature concentrated detergents. HIF-Clear with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in whole mouse brain, is able be used as the evaluation of disease treatment efficiency. also supports expansion microscopy can performed on non-sectioned 15-year-old specimen, well 3-month brain. Thus, represents feasible approach researching archived future neuroscientific 3D neuropathological analyses.

Language: Английский

Citations

0
Revealing intact neuronal circuitry in centimeter-sized formalin-fixed paraffin-embedded brain DOI Creative Commons
Ya‐Hui Lin, Liwen Wang, Yen‐Hui Chen

et al.

eLife, Journal Year: 2024, Volume and Issue: 13

Published: May 22, 2024

Tissue-clearing and labeling techniques have revolutionized brain-wide imaging analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce HIF-Clear, a novel method for efficiently clearing centimeter-thick FFPE specimens using elevated temperature concentrated detergents. HIF-Clear with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in whole mouse brain is able be used as the evaluation of disease treatment efficiency. also supports expansion microscopy can performed on non-sectioned 15-year-old specimen, well 3-month brain. Thus, represents feasible approach researching archived future neuroscientific 3D neuropathological analyses.

Language: Английский

Citations

0
Structural and molecular characterization of astrocyte and vasculature connectivity in the mouse hippocampus and cortex DOI Creative Commons
Charlotte Lorin, Romain Guiet, Nicolas Chiaruttini

et al.

Glia, Journal Year: 2024, Volume and Issue: 72(11), P. 2001 - 2021

Published: July 15, 2024

Abstract The relation of astrocytic endfeet to the vasculature plays a key functional role in neuro‐glia‐vasculature unit. We characterize spatial organization astrocytes and structural aspects that facilitate their involvement molecular exchanges. Using double transgenic mice, we performed co‐immunostaining, confocal microscopy, three‐dimensional digital segmentation investigate biophysical intricate endfoot network at micrometer level isocortex hippocampus. results showed hippocampal had smaller territories, reduced dimensions, fewer contacts with blood vessels compared those isocortex. Additionally, found both connexins 43 30 have higher density former is overexpressed relative latter. However, due limitations method, further studies are needed determine exact localization on endfoot. quantitative information obtained this study will be useful for modeling interactions vasculature.

Language: Английский

Citations

0
A Simple and Fast Optical Clearing Method for Whole‐Mount Fluorescence In Situ Hybridization (FISH) Imaging DOI Creative Commons
Junwoo Suh, Yehe Liu,

Jordan Smith

et al.

Journal of Biophotonics, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 10, 2024

We report a single-step optical clearing method that is compatible with RNA fluorescence in situ hybridization (FISH) imaging. previously demonstrated microscopy imaging immunohistochemistry and genetic reporters using technique called lipid-preserving refractive index matching for prolonged depth (LIMPID). Our protocol reliably produces high-resolution three-dimensional (3D) images minimal aberrations high magnification objectives, captures large field-of-view of whole-mount tissues, supports co-labeling antibody FISH probes. also custom-designed probes quail embryos, demonstrating the ease fabricating use less common animal models. Furthermore, we show high-quality 3D conventional microscope, without more advanced sectioning instruments such as confocal or light-sheet microscopy. For broader adoption, simplified optimized 3D-LIMPID-FISH to minimize barrier entry, provide detailed aid users navigating thick thin

Language: Английский

Citations

0
INSIHGT - Scalable, Accessible, Homogeneous Deep Multiplexed Immunolabelling Platform for 3D Spatial Biology DOI Creative Commons
H. M. Lai,

Jacky Tin Shing Hung,

Chun Ngo Yau

et al.

BIO Web of Conferences, Journal Year: 2024, Volume and Issue: 129, P. 14004 - 14004

Published: Jan. 1, 2024

Language: Английский

Citations

0
A Three Dimensional Immunolabeling Method with Peroxidase-fused Nanobodies and Fluorochromized Tyramide-Glucose Oxidase Signal Amplification DOI Open Access
Kenta Yamauchi, Masato Koike, Hiroyuki Hioki

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 26, 2024

Abstract Three dimensional immunohistochemistry (3D-IHC), immunolabeling of 3D tissues, reveals the spatial organization molecular and cellular assemblies in context tissue architecture. Deep rapid penetration antibodies into tissues highly sensitive detection are critical for high-throughput imaging analysis immunolabeled tissues. Here, we report a nanobody (nAb)-based 3D-IHC, POD-nAb/FT-GO high-speed high-sensitive targets within Peroxidase-fused nAbs (POD-nAbs) enhanced depth allowed by combined with fluorescent tyramide signal amplification system, Fluorochromized Tyramide-Glucose Oxidase (FT-GO). Multiplex labeling was implemented to 3D-IHC quenching POD sodium azide. Using technique, successfully visualized somata processes neuronal glial cells millimeter-thick mouse brain three days. Given its detection, our protocol, would provide useful platform histochemical

Language: Английский

Citations

0
Single-cell synaptome mapping: its technical basis and applications in critical period plasticity research DOI Creative Commons
Motokazu Uchigashima, Takayasu Mikuni

Frontiers in Neural Circuits, Journal Year: 2024, Volume and Issue: 18

Published: Dec. 11, 2024

Our brain adapts to the environment by optimizing its function through experience-dependent cortical plasticity. This plasticity is transiently enhanced during a developmental stage, known as “critical period,” and subsequently maintained at lower levels throughout adulthood. Thus, understanding mechanism underlying critical period crucial for improving adaptability across lifespan. Critical relies on activity-dependent circuit remodeling anatomical functional changes individual synapses. However, it remains challenging identify molecular signatures of synapses responsible understand how these plasticity-related are spatiotemporally organized within neuron. Recent advances in genetic tools genome editing methodologies have enabled single-cell endogenous protein labeling brain, allowing comprehensive profiling neuron, namely “single-cell synaptome mapping.” promising approach can facilitate insights into spatiotemporal organization that sparse yet functionally important single neurons. In this review, we introduce basics mapping discuss applications investigate synaptic cellular mechanisms period.

Language: Английский

Citations

0
Novel Spatial Approaches to Dissect the Lung Cancer Immune Microenvironment DOI Open Access
Idania Lubo, Sharia Hernandez, Ignacio I. Wistuba

et al.

Cancers, Journal Year: 2024, Volume and Issue: 16(24), P. 4145 - 4145

Published: Dec. 12, 2024

Lung cancer is a deadly disease with the highest rates of mortality. Over recent decades, better understanding biological mechanisms implicated in its pathogenesis has led to development targeted therapies and immunotherapy, resulting improvements patient outcomes. To understand lung tumor biology advance towards precision oncology, comprehensive profile necessary. In years, novel situ spatial multiomics approaches have emerged offering more detailed view location microenvironment cells, identifying their unique composition functional status. this sense, platforms been developed evaluate heterogeneity, gene expression, metabolic reprogramming, signaling pathway activation, cell–cell interactions, immune cell programs. research, several studies used these technologies locate cells associated them histological features that are relevant adenocarcinoma. These advancements may unveil further molecular will lead discovery biomarkers for treatment prediction prognosis. review, we provide an overview widely emerging pathology-based profiling how they enhance our response.

Language: Английский

Citations

0