Correlated light and electron microscopy is a powerful approach combining the strengths of fluorescence microscopy and electron microscopy to visualize how protein machineries and cellular ultrastructure change over time.
Fluorescence microscopy (FM) is an ideal way to localize proteins and reveal their dynamics. Even rare or transient events can be identified and the relationship between proteins can be visualized. Structures within the cell, however, remain hidden if they are unlabeled or fall below the diffraction limit of light. Electron microscopy (EM) is ideal for resolving organellar and sub-organellar ultrastructure. In combination with image processing and cryo-EM protein structure can be resolved in situ. However, it is impossible to unambiguously locate rare or transient events in dynamic processes within the vast volume of the cell.
By combining fluorescence and 3D transmission and scanning electron microscopy we can:
- Target the EM acquisition based on fluorescent information.
- Visualize dynamic steps by EM.
- Increase the throughput of EM data acquisition.
- Unambiguous assignment molecular information to cellular ultrastructure.
- Determine the underlying structure of rare, transient or un-described subcellular events.
Overlay image after high precision correlation showing the position of clathrin-coated structures containing fluorescent transferrin (yellow) within the cell.