UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Inner ear tissue preservation by rapid freezing: Improving fixation by high-pressure freezing and hybrid methods.

Bullen, A; Taylor, RR; Kachar, B; Moores, C; Fleck, RA; Forge, A; (2014) Inner ear tissue preservation by rapid freezing: Improving fixation by high-pressure freezing and hybrid methods. Hearing Research , 315 pp. 49-60. 10.1016/j.heares.2014.06.006. Green open access

[thumbnail of Article] PDF (Article)
1-s2.0-S0378595514001178-main.pdf

Download (5MB)

Abstract

In the preservation of tissues in as 'close to life' state as possible, rapid freeze fixation has many benefits over conventional chemical fixation. One technique by which rapid freeze-fixation can be achieved, high pressure freezing (HPF), has been shown to enable ice crystal artefact-free freezing and tissue preservation to greater depths (more than 40μm) than other quick-freezing methods. Despite increasingly becoming routine in electron microscopy, the use of HPF for the fixation of inner ear tissue has been limited. Assessment of the quality of preservation showed routine HPF techniques were suitable for preparation of inner ear tissues in a variety of species. Good preservation throughout the depth of sensory epithelia was achievable. Comparison to chemically fixed tissue indicated that fresh frozen preparations exhibited overall superior structural preservation of cells. However, HPF fixation caused characteristic artefacts in stereocilia that suggested poor quality freezing of the actin bundles. The hybrid technique of pre-fixation and high pressure freezing was shown to produce cellular preservation throughout the tissue, similar to that seen in HPF alone. Pre-fixation HPF produced consistent high quality preservation of stereociliary actin bundles. Optimising the preparation of samples with minimal artefact formation allows analysis of the links between ultrastructure and function in inner ear tissues.

Type: Article
Title: Inner ear tissue preservation by rapid freezing: Improving fixation by high-pressure freezing and hybrid methods.
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.heares.2014.06.006
Publisher version: http://dx.doi.org/10.1016/j.heares.2014.06.006
Language: English
Additional information: © 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).
Keywords: Actin, Electron microscopy, Pre-fixation, Stereocilia, high pressure freezing
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > The Ear Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology
URI: https://discovery.ucl.ac.uk/id/eprint/1434592
Downloads since deposit
83Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item