I will provide an overview of the Correlative Light and Electron Microscopy (CLEM) workflow established in our institute and the different variations possible in our facility. This workflow encompasses the screening of vitrified whole cells on our cryo-confocal microscope, thinning of the cells in a FIB-SEM to a lamella of less than 300 nm, imaging of thin lamella in a cryo-fluorescence microscope and finally cryo-electron tomography on the same lamella in a correlated fashion. I will also give a brief overview of different correlation softwares available for this workflow.
To help one correlate ~300 nm resolution confocal microscopy data and ~3nm resolution cryo electron tomography data, intracellular fiducials may be helpful. Examples are gold particles, intracellular dyes and molecular markers. I will give a short overview of several of these strategies and talk about advantages and disadvantages.
The Joint Lab Model Driven Materials Characterization is a cross-centre platform of the Helmholtz Association.