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

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Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue. / Studer, Daniel; Zhao, Shanting; Chai, Xuejun; Jonas, Peter; Graber, Werner; Nestel, Sigrun; Frotscher, Michael.

In: NAT PROTOC, Vol. 9, No. 6, 01.01.2014, p. 1480-95.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Studer, D, Zhao, S, Chai, X, Jonas, P, Graber, W, Nestel, S & Frotscher, M 2014, 'Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue', NAT PROTOC, vol. 9, no. 6, pp. 1480-95. https://doi.org/10.1038/nprot.2014.099

APA

Studer, D., Zhao, S., Chai, X., Jonas, P., Graber, W., Nestel, S., & Frotscher, M. (2014). Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue. NAT PROTOC, 9(6), 1480-95. https://doi.org/10.1038/nprot.2014.099

Vancouver

Studer D, Zhao S, Chai X, Jonas P, Graber W, Nestel S et al. Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue. NAT PROTOC. 2014 Jan 1;9(6):1480-95. https://doi.org/10.1038/nprot.2014.099

Bibtex

@article{8ad9268887464bee8b8371c68ea29def,
title = "Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue",
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.",
keywords = "Brain, Cryoelectron Microscopy, Neurons, Pressure, Synapses",
author = "Daniel Studer and Shanting Zhao and Xuejun Chai and Peter Jonas and Werner Graber and Sigrun Nestel and Michael Frotscher",
year = "2014",
month = jan,
day = "1",
doi = "10.1038/nprot.2014.099",
language = "English",
volume = "9",
pages = "1480--95",
journal = "NAT PROTOC",
issn = "1754-2189",
publisher = "NATURE PUBLISHING GROUP",
number = "6",

}

RIS

TY - JOUR

T1 - Capture of activity-induced ultrastructural changes at synapses by high-pressure freezing of brain tissue

AU - Studer, Daniel

AU - Zhao, Shanting

AU - Chai, Xuejun

AU - Jonas, Peter

AU - Graber, Werner

AU - Nestel, Sigrun

AU - Frotscher, Michael

PY - 2014/1/1

Y1 - 2014/1/1

N2 - 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.

AB - 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.

KW - Brain

KW - Cryoelectron Microscopy

KW - Neurons

KW - Pressure

KW - Synapses

U2 - 10.1038/nprot.2014.099

DO - 10.1038/nprot.2014.099

M3 - SCORING: Journal article

C2 - 24874814

VL - 9

SP - 1480

EP - 1495

JO - NAT PROTOC

JF - NAT PROTOC

SN - 1754-2189

IS - 6

ER -