Condensed Matter Seminar: "Long Term 3D Imaging by FIBSEM for Neurons and Cell Biology and Correlation to Cryo Fluorescence Microscopy"

Harald Hess (Howard Hughes Medical Institute, Janelia Research Campus)
- | David Rittenhouse Laboratory, A4

3D Electron microscopy volume data can be acquired by a variety of approaches.  Focused Ion beam – scanning electron microscopy, FIBSEM, offers no limitation on section thickness, so that isotropic voxels with 8 nm or less sampling in x,y,z dimensions can be acquired.  This capability opens a new regime where entire cells can be imaged with 4 nm voxel sampling, thereby surpassing partial cell or section limitations to complete cell data. The heavy metal staining for EM contrast gives spatially detailed but generic black and white rendering of protein and membrane defined structures.  On the other hand, fluorescence microscopy is highly protein specific, by labeling only a tiny subset (1-3) of the thousands of constituent proteins of the cell. Correlated cryogenic fluorescence microscopy offers a way to combine both without compromising the quality of either EM or fluorescence image.