ArticleCell reports methods2023
Preserving extracellular space for high-quality optical and ultrastructural studies of whole mammalian brains.
Article in Cell reports methods, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Review
- Probing molecular diversity and ultrastructure of brain cells with fluorescent aptamers.Nature communications · 2026Article
- Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues.Nature communications · 2025Article
- Article
- Multiplexed volumetric CLEM enabled by scFvs provides insights into the cytology of cerebellar cortex.Nature communications · 2024Article
- Dense, Continuous Membrane Labeling and Expansion Microscopy Visualization of Ultrastructure in Tissues.bioRxiv : the preprint server for biology · 2024Article
- Volume electron microscopy in injured rat brain validates white matter microstructure metrics from diffusion MRI.Imaging neuroscience (Cambridge, Mass.) · 2024Article
- Astrocyte coverage of excitatory synapses correlates to measures of synapse structure and function in primary visual cortex.bioRxiv : the preprint server for biology · 2023Article
- Multiplexed volumetric CLEM enabled by antibody derivatives provides new insights into the cytology of the mouse cerebellar cortex.Research square · 2023Article
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6 authors.
Funding
Abstract
Analysis of brain structure, connectivity, and molecular diversity relies on effective tissue fixation. Conventional tissue fixation causes extracellular space (ECS) loss, complicating the segmentation of 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 in the hydrostatic pressure within the vasculature. A more reliable fixation protocol that uniformly preserves the ECS throughout whole brains would greatly benefit a wide range of neuroscience studies. Here, we report a straightforward transcardial perfusion strategy that preserves ECS throughout the whole rodent brain. No special setup is needed besides sequential solution changes, and the protocol offers excellent reproducibility. In addition to better capturing tissue ultrastructure, preservation of ECS has many downstream advantages such as accelerating heavy-metal staining for electron microscopy, improving detergent-free immunohistochemistry for correlated light and electron microscopy, and facilitating lipid removal for tissue clearing.
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