Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography

Anna Bieber; Cristina Capitanio; Florian Wilfling; J&#252;rgen Plitzko; Philipp S. Erdmann

doi:10.3791/62886

Journal

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Biology

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Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography

JoVE Journal
Biology

This content is Free Access.

JoVE Journal Biology

Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography

3,333 Views

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08:20 min

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October 25, 2021

DOI:

10.3791/62886-v

DOI:

10.3791/62886-v

•

08:20 min

October 25, 2021

•

3316 Views

Anna Bieber*¹, Cristina Capitanio*¹, Florian Wilfling^1,2, Jürgen Plitzko¹, Philipp S. Erdmann^1,3

¹Max Planck Institute of Biochemistry ²Max Planck Institute for Biophysics ³Fondazione Human Technopole

Summary

Here, we present a pipeline for 3D-correlative focused ion beam milling on guiding the preparation of cellular samples for cryo-electron tomography. The 3D position of fluorescently tagged proteins of interest is first determined by cryo-fluorescence microscopy, and then targeted for milling. The protocol is suitable for mammalian, yeast, and bacterial cells.