iMEM: Isolation of Plasma Membrane for Cryoelectron Microscopy.

Peitsch, Camille; Beckmann, Sven; Zuber, Benoît (2016). iMEM: Isolation of Plasma Membrane for Cryoelectron Microscopy. Structure, 24(12), pp. 2198-2206. Cell Press 10.1016/j.str.2016.09.016

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The plasma membrane and the cell cortex are essential parts of the eukaryotic cell. The plasma membrane delimitates the cell and mediates communication with the outside. The cell cortex is the submembrane cytoskeleton shaping the cell and is able to reorganize for the passage of material. To study events at and near the plasma membrane, cryoelectron microscopy (cryo-EM) may be used. Most intact cells are too thick for direct cryo-EM imaging. Generating cell-free membrane patches could be a means to study features at the plasma membrane. Here we present an unroofing method, termed iMEM (isolation of membrane patches for cryo-EM) where the plasma membrane is isolated directly on an EM grid. The in situ isolation of membrane patches has several advantages: it is a one-step procedure providing a higher throughput than focused-ion beam cryomilling. It enables the time-precise control over biochemical events before cryofixation.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Anatomy
09 Interdisciplinary Units > Microscopy Imaging Center MIC

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Peitsch, Camille; Beckmann, Sven and Zuber, Benoît

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0969-2126

Publisher:

Cell Press

Language:

English

Submitter:

Benoît Zuber

Date Deposited:

12 Dec 2016 10:45

Last Modified:

14 Feb 2019 18:01

Publisher DOI:

10.1016/j.str.2016.09.016

PubMed ID:

27818102

Uncontrolled Keywords:

actin cortex; blotting; cell-free system; cryoelectron tomography; electron microscopy; endocytosis; exocytosis; filter paper; large dense core vesicles; live fluorescence imaging; plasma membrane

BORIS DOI:

10.7892/boris.91356

URI:

https://boris.unibe.ch/id/eprint/91356

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