A new tool based on two micromanipulators facilitates the handling of ultrathin cryosection ribbons

Studer, Daniel Franz; Klein, Alycia; Iacovache, Mircea Ioan; Gnaegi, Helmut; Zuber, Benoît (2014). A new tool based on two micromanipulators facilitates the handling of ultrathin cryosection ribbons. Journal of structural biology, 185(1), pp. 125-128. Elsevier 10.1016/j.jsb.2013.11.005

[img]
Preview
Text
Studer_Micromanipulators_2014.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (1MB) | Preview

A close to native structure of bulk biological specimens can be imaged by cryo-electron microscopy of vitreous sections (CEMOVIS). In some cases structural information can be combined with X-ray data leading to atomic resolution in situ. However, CEMOVIS is not routinely used. The two critical steps consist of producing a frozen section ribbon of a few millimeters in length and transferring the ribbon onto an electron microscopy grid. During these steps, the first sections of the ribbon are wrapped around an eyelash (unwrapping is frequent). When a ribbon is sufficiently attached to the eyelash, the operator must guide the nascent ribbon. Steady hands are required. Shaking or overstretching may break the ribbon. In turn, the ribbon immediately wraps around itself or flies away and thereby becomes unusable. Micromanipulators for eyelashes and grids as well as ionizers to attach section ribbons to grids were proposed. The rate of successful ribbon collection, however, remained low for most operators. Here we present a setup composed of two micromanipulators. One of the micromanipulators guides an electrically conductive fiber to which the ribbon sticks with unprecedented efficiency in comparison to a not conductive eyelash. The second micromanipulator positions the grid beneath the newly formed section ribbon and with the help of an ionizer the ribbon is attached to the grid. Although manipulations are greatly facilitated, sectioning artifacts remain but the likelihood to investigate high quality sections is significantly increased due to the large number of sections that can be produced with the reported tool.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Anatomy

UniBE Contributor:

Studer, Daniel Franz; Iacovache, Mircea Ioan and Zuber, Benoît

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1047-8477

Publisher:

Elsevier

Funders:

[4] Swiss National Science Foundation

Language:

English

Submitter:

Benoît Zuber

Date Deposited:

20 Mar 2014 20:32

Last Modified:

09 Dec 2014 00:27

Publisher DOI:

10.1016/j.jsb.2013.11.005

PubMed ID:

24269483

Uncontrolled Keywords:

Cryo-electron microscopy Cryosectioning Electron microscopy Frozen-hydrated sections High pressure freezing Micromanipulation Ultramicrotomy

BORIS DOI:

10.7892/boris.46869

URI:

https://boris.unibe.ch/id/eprint/46869

Actions (login required)

Edit item Edit item
Provide Feedback