ArticleNature protocols2014
Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue.
Article in Nature protocols, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Pressure enhanced dual-solid-surface ultra-rapid cooling improves post-thaw recovery in hepatocytes and precision cut liver slices.Scientific reports · 2026Article
- Cryogenic single-molecule fluorescence imaging.BMB reports · 2025Review
- Presynaptic cAMP-PKA-mediated potentiation induces reconfiguration of synaptic vesicle pools and channel-vesicle coupling at hippocampal mossy fiber boutons.PLoS biology · 2024Article
- The desmosome comes into focus.The Journal of cell biology · 2024Review
- Disruption of the mitochondrial network in a mouse model of Huntington's disease visualized by in-tissue multiscale 3D electron microscopy.Acta neuropathologica communications · 2024Article
- Article
- In situ label-free X-ray imaging for visualizing the localization of nanomedicines and subcellular architecture in intact single cells.Nature protocols · 2024Review
- Cryopreservation of brain cell structure: a review.Free neuropathology · 2024Article
- A Compartmentalized Neuronal Cell-Culture Platform Compatible With Cryo-Fixation by High-Pressure Freezing for Ultrastructural Imaging.Frontiers in neuroscience · 2021Article
- Ultrastructural Imaging of Activity-Dependent Synaptic Membrane-Trafficking Events in Cultured Brain Slices.Neuron · 2020Article
- Pathology of myelinated axons in the PLP-deficient mouse model of spastic paraplegia type 2 revealed by volume imaging using focused ion beam-scanning electron microscopy.Journal of structural biology · 2020Article
- Functional Electron Microscopy, "Flash and Freeze," of Identified Cortical Synapses in Acute Brain Slices.Neuron · 2020Article
- Ultrastructural Correlates of Presynaptic Functional Heterogeneity in Hippocampal Synapses.Cell reports · 2020Article
- Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells.Science (New York, N.Y.) · 2020Article
- Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2018Article
- Correlating Fluorescence and High-Resolution Scanning Electron Microscopy (HRSEM) for the study of GABAScientific reports · 2017Article
- Array tomography of physiologically-characterized CNS synapses.Journal of neuroscience methods · 2016Article
- Energy and Potassium Ion Homeostasis during Gamma Oscillations.Frontiers in molecular neuroscience · 2016Review
- Fine structure of synapses on dendritic spines.Frontiers in neuroanatomy · 2014Review
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Electron microscopy (EM) allows for the simultaneous visualization of all tissue components at high resolution. However, the extent to which conventional aldehyde fixation and ethanol dehydration of the tissue alter the fine structure of cells and organelles, thereby preventing detection of subtle structural changes induced by an experiment, has remained an issue. Attempts have been made to rapidly freeze tissue to preserve native ultrastructure. Shock-freezing of living tissue under high pressure (high-pressure freezing, HPF) followed by cryosubstitution of the tissue water avoids aldehyde fixation and dehydration in ethanol; the tissue water is immobilized in ∼50 ms, and a close-to-native fine structure of cells, organelles and molecules is preserved. Here we describe a protocol for HPF that is useful to monitor ultrastructural changes associated with functional changes at synapses in the brain but can be applied to many other tissues as well. The procedure requires a high-pressure freezer and takes a minimum of 7 d but can be paused at several points.
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