Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron TomographyAnna Bieber *1, Cristina Capitanio *1, Florian Wilfling 1,2, Jürgen Plitzko 1, Philipp S. Erdmann 1,3
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.
