CLEM workflow options
The Wolfson Bioimaging Facility houses specialised equipment suitable for CLEM experiments.
- Cells containing fluorescent tracers grown on sapphire disks can be imaged by fluorescence microscopy techniques then rapidly frozen using our Leica EMPACT2 and rapid transfer system (RTS) for subsequent embedding and sectioning. The EMPACT2/RTS system is mobile and so can be positioned beside most of our fluorescence microscopes to enable freezing of cells within 5 seconds of the termination of live cell imaging.
- Fluorescent markers in cryo-fixed material can be imaged using our cryo-fluorescent microscope, which maintains the specimen in a liquid nitrogen-cooled state prior to cryo-EM. The cryo-fluorescence microscope is a Leica DM6000 with DFC365FX camera and Leica Cryo CLEM system.
- Our photo-oxidation microscope system is designed for experiments with mini-SOG, a genetically-encoded fluorescent tag which also generates singlet oxygen. This can be used to mark the location of the fluorescent protein with electron-dense DAB deposit. The photo-oxidation microscope is a Leica DM4000 with DFC365FX camera and a xenon light source to initiate photo-oxidation.
If you are considering using this equipment for CLEM experiments, we advise contacting Paul Verkade to discuss technical details.