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

DOI :

10.3791/62886-v

8:20 min

October 25th, 2021

October 25th, 2021

2,471 Views

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

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.

Tags

3D correlative FIB Milling

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