Evan Krystofiak is a staff scientist in the Cell Imaging Shared Resource (CISR) core facility at Vanderbilt University, performing electron microscopy for over 90 labs using a combination of conventional and cryo-based approaches. Dr. Krystofiak’s main research interests lie in understanding macromolecular ultrastructure in the in-situ cellular context. His research includes development of high-resolution freeze-etch using amorphous replicas, freeze-substitution of difficult to preserve samples, and Fourier space analysis to understand biological paracrystal packing. He has used these techniques to examine the structure of tight junction fibrils and revealed the hydrated structure of enterocyte glycocalyx.
- Outline the basic methodology of Freeze-Fracture and Freeze-Etch techniques
- Discuss the role of the enterocyte glycocalyx and similar extracellular structures in situ
- Illustrate how to optimize these replica techniques for maximum biological detail