Associate Professor, Monash University, Dept. of Biochemistry and Molecular Biology, ARC Centre of Excellence in Advanced Molecular Imaging, Clayton, AUSTRALIA

Thursday, 16 June 2022 | 12:00 noon – ICGEB Trieste, ITALY

Increasing throughput and efficiency in cryo-CLEM

Host: E. Buratti

Cryo-transmission electron tomography (cryoET) associated with cryo-focused ion beam (cryoFIB) milling enables structural biology studies to be performed directly within the cellular environment. Cryo- preserved cells are milled, and a lamella with a thickness of 200-300 nm provides an electron transparent window suitable for cryoET imaging. CryoFIB milling is an effective method, but it is tedious and time- consuming. Even with the recent advances in automation, when the average diameter of a cell is over 10 um, the lamella only represents a minimal fraction of the sample. Since most cellular processes occur in well-defined regions, precise targeting is critical to success. Here we present a new generation of integrated cryo Correlative Light and Electron Microscope (cryo-CLEM) the PIE-scope2, capable of super- resolution fluorescence imaging. A new user interface allows a smoother integration with targeted on- grid lamella preparation and the possibility of isolating sub-micrometre regions from large tissue.

Alex completed a PhD in biophysics at EMBL Heidelberg in the group of John Briggs to develop image processing methods using cryo-electron tomography. His work resulted in the first structural description of the viral maturation in HIV-1 and the first sub-nm structure ever solved using sub-tomographic averaging. After graduation he joined FEI Company (now ThermoFisher) where he was responsible for the development of hardware and software for correlative microscopy and nanometre resolution volume imaging. In 2015 he joined Monash University as Associate professor. Here his group develops methods and technology for molecular imaging and visual proteomics.

#ICGEBseminars #ICGEBalumni

Hundreds of videos, tens of playlists providing free scientific content worldwide