Structural Biology and Biochemistry (SBB) Theme Seminar

Title: The application of Cryogenic Correlative Light and Electron Microscopy to Protein Trafficking

Speaker: Dr. Nick Ariotti, IMB

Bio: Dr Ariotti is a cell biologist specialising in how defects in protein trafficking, and plasma membrane organisation can result in human diseases. He completed his PhD in 2013 at the University of Queensland. He then spent 4 years postdoctoral researcher at the Institute for Molecular Biosciences in Robert Parton’s laboratory where he focused on developing and applying correlative light and electron microscopy approaches to answer questions regarding endocytosis. In 2017, Dr Ariotti was recruited to the Electron Microscope Unit at UNSW to serve as the Associate Director of Biological EM. He spent 5 years at the Electron Microscope Unit establishing Cryo-EM and cryogenic-Correlative Light and Electron Microscopy. In 2022, Dr Ariotti returned to the IMB at UQ as an independent research group leader with a focus on in situ cryo-correlative light and electron microscopy. He has published seminal research in Nature, Nature Materials, Nature Cell Biology, Nature Nanotechnology, Cell, and Developmental Cell.

Abstract: in situ cryo-EM has the potential to bridge structural biology and cellular biology to provide unparalleled detail about how proteins, macromolecular complexes and organelles interact with each other to illicit their cellular functions. Despite this promise, in situ cryo-EM remains an under-represented workflow in the field. The current state-of-the-art poses significant challenges with respect to inherent limitations in the sample thickness, challenges with engineering effective sample transfer solutions between disparate instruments and automation challenges that impede data acquisition at scale. This talk will address where the field is heading with in situ structural biology, discuss ways to overcome sample bottlenecks, and provide a simplified in situ cryo-EM alternative for researchers looking to transition into this advanced workflow. I will discuss optimal substrate parameters for improved yield of areas for high-quality in situ cryo-EM imaging and characterise ideal cell type properties for high quality imaging. We show that these improvements can be utilised for sub-tomogram averaging macromolecular complexes in situ. Finally, we explore how cryo-correlative light and electron microscopy can be utilised for in situ cryo-EM of analyses of complex biological processes such as protein trafficking.