Studer, Daniel Franz; Zhao, Shanting; Chai, Xuejun; Jonas, Peter; Graber, Werner Adrian; Nestel, Sigrun; Frotscher, Michael (2014). Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue. Nature protocols, 9(6), pp. 1480-1495. Nature Publishing Group 10.1038/nprot.2014.099
Full text not available from this repository. (Request a copy)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.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Anatomy |
UniBE Contributor: |
Studer, Daniel Franz, Graber, Werner Adrian |
Subjects: |
600 Technology > 610 Medicine & health |
ISSN: |
1754-2189 |
Publisher: |
Nature Publishing Group |
Language: |
English |
Submitter: |
Werner Adrian Graber |
Date Deposited: |
10 Oct 2014 22:07 |
Last Modified: |
05 Dec 2022 14:35 |
Publisher DOI: |
10.1038/nprot.2014.099 |
PubMed ID: |
24874814 |
URI: |
https://boris.unibe.ch/id/eprint/53893 |
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