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
I am a seasoned electron microscopist and imaging scientist. My experience ranges from cryo-Transmission Electron Microscopy (cryo-TEM) of soft matter systems (polymers, proteins, liposomes, extra cellular vesicles) to focused ion beam scanning electron microscopy (FIB-SEM), and particularly the development of cryo-FIB-SEM sample preparation approaches applied to soft matter systems. I have been developing the cryogenic lift-out technique since 2013 to enable cryo-TEM of bulk samples (polymers, cells, tissues). At the University of Nottingham, I am the manager of the Cryogenic Analytical and Transfer (CAT) Suite, a category 2 approved microscopy suite with an emphasis on cryogenic and correlative (light and electron) microscopy. In 2024 I will add confocal light microscopy to my skills and responsibilities in the lab. I believe that I have a wide range of skills and backgrounds to offer the RMS gained from my years in the lab and also developing training and organizational skills to fulfil my various roles. I have so far contributed to the running of two cryo-EM courses in conjunction with the RMS (2021, fully virtual and 2023 as a hybrid). I have a keen interest in the development and training of the next generation of microscopy professionals.
I am a board member of Journal of Microscopy and since 2022 is the Chair of the Cryo Microscopy Group. Since 2014 I have been the editor of Microscopy and Analysis, a global microscopy publication aimed at the professional microscopy community. I became Editor-in-Chief of M&A in 2020. I believe that this has given me a good appreciation of what is going on in microscopy, of new developments and where the field is headed. I enjoy observing this and trying to keep ahead of the curve.
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.