Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue.

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

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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 and 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:

11 May 2016 08:57

Publisher DOI:

10.1038/nprot.2014.099

PubMed ID:

24874814

URI:

https://boris.unibe.ch/id/eprint/53893

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