Correlative light and
electron microscopy
Combining the power of light and electron microscopy
Combine the resolving power of electron microscopy with the labelling possibilities of fluorescence microscopy
Resolving fine details at the organelle level is a difficult challenge that often requires a resolution only electron microscopes can achieve but their use is limited to fixed tissues and labelling specific cell compartments is difficult. Correlative light and electron microscopy combines both imaging tools to bypass this limitation and can be used to trace and reconstruct cellular structures in 3D to show the tissue context.
Gaining high resolution insight into cell interaction with other compartments in 3D
Combining high-resolution confocal microscopy with serial blockface scanning electron microscopy enables studying complex cell-cell interactions in the tissue context using fluorescence for specifically labelling cells of interest and electron microscopy to resolve them at the organelle level. Bonardel and colleagues used this technique to map the cellular interactions imprinting the identity of Kupffer cells and gain unparalleled insights into their niche organization
Imaging intracellular aggregation and phase separation with 3D array tomography
The phase separation of FLOE1-GFP fusion protein is shown in the cellular context of a U2OS cell for an ROI tracked over 21 sections.
Live super resolution correlative imaging
In addition to our established acquisition and correlation pipeline, we aim to uniquely integrate live super-resolution, SIM2 with EM. We leverage NIRB to will allow us to correlate ROIs at a resolution of 60nm (SIM2) down to <20nm (PALM/dSTORM) with ultrastructure in 3D.
This extends the functionality of our focused ion beam scanning electron microscope (FIB-SEM), itself a spearpoint technique in Flanders.
Examples of 3D-FIB-SEM.
A. SIM projection images from GFP-PSEN2 expressing cell (green) labelled with MitoTracker (red)
B. FIB-SEM of square in A
C. Rectangle area segmentation from B: LE/Lys (purple), mitochondria (orange), LD (light orange), nucleus (green) D. 3D reconstruction
E. Visible fusion/fission of tubular structure
