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Published on: February 27, 2020
iMEM: Isolation of Plasma Membrane for Cryoelectron Microscopy
Camille Françoise Peitsch1, Sven Beckmann2, Benoît Zuber2
1Experimental Morphology, Institute of Anatomy, University of Bern, Baltzerstrasse 2, Bern 3012, Switzerland; Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern 3012, Switzerland.
Researchers developed iMEM, a new method to isolate plasma membrane patches for cryo-electron microscopy (cryo-EM). This technique allows detailed study of cell surface events and offers higher throughput than existing methods.
Area of Science:
- Cell biology
- Microscopy techniques
Background:
- The plasma membrane and cell cortex are vital for eukaryotic cell structure and function.
- Studying plasma membrane events using cryo-electron microscopy (cryo-EM) is challenging due to cell thickness.
Purpose of the Study:
- To present a novel method for isolating plasma membrane patches suitable for cryo-EM.
- To overcome limitations of existing techniques for visualizing sub-membrane structures.
Main Methods:
- Developed iMEM (isolation of membrane patches for cryo-EM), a one-step unroofing technique.
- Plasma membrane isolation directly on an EM grid.
- Enables time-controlled biochemical manipulation before cryofixation.
Main Results:
- iMEM provides a higher throughput method compared to focused-ion beam cryomilling.
- Facilitates in situ isolation of membrane patches for cryo-EM analysis.
- Allows precise temporal control over biochemical events prior to cryofixation.
Conclusions:
- iMEM is an efficient method for preparing plasma membrane samples for cryo-EM.
- This technique advances the study of cell surface dynamics and interactions.
- Enables detailed structural analysis of the plasma membrane and associated cortical structures.
