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

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

Article DOI: 10.3791/62886-v • 08:20 min • October 25th, 2021
October 25th, 2021 •
*1, *1, 1,2, 1, 1,3
1Max Planck Institute of Biochemistry, 2Max Planck Institute for Biophysics, 3Fondazione Human Technopole
* These authors contributed equally

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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.

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3D-correlative FIB Milling High-resolution Cryo-electron Tomography Specific Cellular Events Native Environment Ultra Structure Correlative Workflow Rare Cellular Structures Cryo-fluorescence Microscopy Grid Squares FIB-SEM TEM Instrument Fluorescent Stack Numerical Aperture Objectives Confocal Microscope Liquid Nitrogen
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