Corporate Member Hosted Event
21 March 2024
09:00 - 10:00 GMT & 15:00 - 16:00 GMT
Cryo-focused ion beam (FIB) micromachining of frozen-hydrated cells has provided new and exciting ways to study their inner workings at the ultrastructural and molecular scale. When combined with cryo-electron tomography (cryo-ET), this method allows us to explore large macromolecular structures within their native and functional cellular environment. Until recently, this technique has mostly been applied to studies on individual cells that are effectively vitrified by plunge-freezing and routinely thinned by cryo-FIB, leaving more complex multicellular and tissue biology out of reach. The application of cryo-lift-out now enables the production of vitrified lamellae from high-pressure frozen samples, encompassing 3D cell cultures, whole small model organisms and tissues. The key step in this advanced preparation entails a micromanipulator system cooled to deep cryogenic temperature that is used to extract targeted portions from vitrified voluminous specimens and make them amenable to subsequent cryo-ET imaging.
An Oxford Instruments webinar,
hosted by the Royal Microscopical Society.
In this webinar, you will learn from users about their challenges, solutions and experiences related to cryo-lift-out workflows in context of cryo-correlative light and electron microscopy. This includes discussion about technical solutions, using the OmniProbe 350 cryo and challenges related to handling and transfer of liftout samples from the FIB-SEM.
Who should attend?
Anyone interested in cryo-FIB sample preparation and analysis of cryogenic samples
You will learn
1. How sample preparation techniques such as site specific lift-out can be used in combination with cryo-FIB
2. Status of current solutions offered for cryo lift-out
3. Practical issues to consider in relation to cryo lift-out
Presenters:
Kim Larsen
Kim Larsen has been with Oxford Instruments for 20 years, during this time Kim has worked in several areas including Customer Support, Development and Marketing. He is currently working in the Product Management team as Product Manager with focus on FIB, OmniProbe and EBSD.
Edoardo D'Imprima
Edoardo D'Imprima coordinates the upcoming Correlative Light and Electron Microscopy Core at the Humanitas Research Hospital, Rozzano (Italy). During his postdoc at EMBL, he investigated cancer onset in tissues using organoid 3D-cell culture systems. For that, together with industrial partners, he developed a multimodal imaging pipeline to link live cell imaging, CLEM, cryo-FIB/SEM volume imaging, lift-out and cryoET. He completed his PhD at Max Planck Institute of Biophysics, focusing on the structural biology of isolated membrane proteins by single particle cryoEM. Before graduating, Edoardo worked as a staff scientist for Merck Serono Italia.
Evgenia Zagoriy
Evgenia Zagoriy is an expert in cryo-focused ion beam (FIB) preparations of vitrified biological specimens, in situ cryo-electron tomography and cryo-correlative light and electron microscopy (CLEM). She has been working in the group of Julia Mahamid in the Structural and Computational Biology Unit at the European Molecular Biology Laboratory Heidelberg (Germany) since 2017. During her postdoctoral tenure at the Max Planck Institute of Biochemistry and her PhD at the Max Planck Institute of Cell Biology and Genetics, she investigated chromosome biology and the regulation of cell division using state-of-the-art fluorescence light microscopy and biochemical methods. Prior to her doctoral studies, she obtained a Master's degree in Biochemistry at the Faculty of Chemistry of the Aristotle University of Thessaloniki (Greece).
Chris Parmenter
Christopher is a Research Officer in Cryogenic Electron Microscopy. His PhD is in polymer chemistry through which he developed skills in Cryo-TEM whilst investigating soft and biological systems. He recently moved into SEM, specifically Cryo-SEM and FIB-SEM of any samples where there is a nano-scale problem to solve or understand. He is keen to meet with other microscopists to further his knowledge of EM & other techniques.
Chris is currently the Editor of Microscopy & Analysis.
RMS Contact:
Victoria Masters
victoria@rms.org.uk
01865 254769
Contact Victoria for RMS Event Hosting, Corporate Member Hosted events and RMS EDI&A queries.