From Correlative to 3D – Recent Advances in Bio Scanning Electron Microscopy
Irani Hall RRI 101
Kirk J. Czymmek, Ph.D.
Director of North American Applications & Labs
Carl Zeiss Microscopy, LLC
Biological structures, such as organelles, bacteria, somatic cell layers, tissues and model organisms are inherently three-dimensional (3D) and technologies that aid visualization and analysis of 3D biological samples is growing rapidly. Specifically, recent developments in Bio Scanning Electron Microscopy allow high-resolution imaging of samples embedded in resin, then physically sliced and the resulting sections collected in a serial ribbon on tape, glass slide or slotted grid. Each approach has unique benefits and allows access to antibody and other affinity probes for interrogation of specific molecules. Alternatively, 3D tomograms of entire sliced volumes can be automated with serial block face imaging, where an ultramicrotome is either integrated directly into the electron microscope chamber or a focused ion beam (FIB) is used to mill away the specimen. Block face images have significantly lower distortions associated with material removal than the compression seen in thin sections, thus final 3D EM reconstructions from this approach align closely with the 3D LM volume image. Furthermore, a far greater understanding of the structure-function relationship in cells and tissues is now achievable via high resolution correlation of chemical markers and structural components in both 2D and 3D dimension. This presentation will focus on contemporary methods and probes, methods and workflow for array tomography, serial block face imaging and correlative light and electron microscopy (CLEM) and demonstrate with practical examples how these tools can provide powerful and novel insights into biological problems.
If you have any questions and to RSVP please contact your Zeiss Electron Microscopy Specialist, KD Derr, at firstname.lastname@example.org.
Refreshments will be served.