Single Particle Cryo-TEM School

10 – 13 December 2018 + Add to calendar Leeds

Single Particle Cryo-TEM School

10 – 13 December 2018 + Add to calendar Leeds

This course provides an introduction to the single particle cryo-TEM pipeline for solving high resolution structures of macromolecular complexes. The 4 day course focuses on providing hands on training for sample preparation by plunge freezing (Vitrobot Mk 4, Leica EM GP) and loading into Titan Krios microscopes, as well as demonstrations of high pressure freezing and non-blotting (spraying) freezing devices. With small group (3 participants) teaching, delegates will get a day on the Titan Krios microscopes to become familiar with imaging/data collection using a Titan Krios microscope with EPU software. Course participants have the option to bring their own samples to prepare grids, and may have a limited opportunity to image and collect data during the course. We welcome course participants with a range of experience and backgrounds, including from academia and industry.

This course is now full, however if you would like to be added to the waiting list please email [email protected]

Outline Schedule

This is the outline schedule for the course. A more detailed programme will be available soon.

The course will begin on Monday 10 December at 09.00 and finish on Thursday 13 December at 14.30. You can bring your own samples to work on during the course.

Day1

Lectures on:

Building projects up for Cryo-EM
Cryo-EM Sample Prep
Emerging Technologies for Sample Prep
Microscope Hardware
Single Particle TEM/Low Dose Imaging
Electron Tomography

Day 2-4

Five half day Practical Sessions including two sessions on Titan Krios microscopes, and vitrification using the Vitrobot Mk 4, EM GP, and other freezing instruments (non blotting devices, high-pressure freezing)

Day 4

Half day of lectures on Image Processing

Organiser and Speaker List

Organiser

Dr Rebecca Thompson

Electron Microscopy Deputy Chair

University of Leeds
Rebecca is Facility Manager and senior cryo-electron microscopy (EM) support scientist at the Astbury Biostructure Laboratory, University of Leeds. Her research interests include imaging a broad range of biological specimens, from whole cells to macromolecular complexes, to high resolution using cryo-EM, and integrating data from EM with other microscopy techniques.

Confirmed Speakers

Mr Alan Boswell

Thermo Fisher
Alan is an experienced senior field service engineer for Thermo Fisher Scientific, formerly FEI. With over 30 years’ experience working with transmission electron microscopes, Alan has specialised in installing and maintaining Titan Krios systems since their introduction into the market ten years ago. He mentors trainee engineers using his teaching skills, and is regularly asked to contribute to product development and projects improving the customer experience. He has recently accepted the challenge of service manager for Thermo Fisher TEM life science within the UK.
Dr Dan Clare

Diamond Light Source
I am Senior Beamline Scientist at the electron-Bioimaging Centre (eBIC) the UK’s national cryo-electron microscopy (cryo-EM) facility based at Diamond light source. My role at eBIC involves co-managing the cryo-EM facility and the international user program that we offer on our four Titan Krios microscopes. Before eBIC I was a senior post-doc in the laboratory of professor Helen saibil at Birkbeck College in London. I am a structural biologist by training and have had a longstanding interest in using cryo-EM and single particle analysis to look at how molecular chaperones function. I also have an active interest in cryo-electron tomography as a way of structurally charactering more complicated systems and currently have a research project looking at the bacterial infection of mammalian cells.
Dr Juan Fontana

University of Leeds
Juan Fontana has over 13 years of experience in structural virology and electron microscopy (EM). During his PhD (CNB-CSIC, Madrid, Spain), he characterised the replication complexes of Bunyamwera and Rubella viruses. This work allowed him to elucidate the mechanisms by which viral macromolecular complexes interact with cellular components to create the architectures of viral factories and to propose novel working models for each replication complex. As a post-doc (LSBR, NIH, Maryland, USA), he worked on Herpes Simplex Virus (HSV) & Influenza entry and on HIV & HSV morphogenesis. These projects included, 1) Elucidating the mechanism of Herpes Simplex Virus (HSV) neutralization by antibodies directed at the fusion domains of HSV’s fusion protein. 2) Unraveling structural intermediates of Influenza Virus fusion protein and M1 matrix protein prior to viral fusion. 3) Describing the two-step fusion process of the retrovirus Avian Sarcoma/Leukosis, in which receptor binding necessarily precedes low-pH activation of the fusion protein. 4) Studying the effects of several drugs and mutations that affect HIV maturation, an essential process for viral infectivity that involves proteolysis of precursor proteins by the viral protease and assembly of a conical core. Overall these projects gave Juan an in-depth knowledge of cell culture, manipulation of viruses, fluorescence & confocal microscopy, and cellular EM; and an expert knowledge of cryo-EM, including sample preparation, data acquisition, high-resolution single-particle reconstruction & atomic modeling and cryo-electron tomography & subtomogram averaging.

On April 2016, Juan started a tenure-track academic fellowship at the University of Leeds. His lab uses cryo-EM, combined with other structural approaches, to characterise macromolecular assemblies, and viruses that are a current public health risk, like HIV, Influenza, herpesviruses and bunyaviruses.
Dr Emma Hesketh

University of Leeds
Emma joined the university of Leeds in 2015 as a postdoctoral researcher in Neil Ranson's group. During this time her research interests focused on the structure of viruses using cryo-electron microscopy to preform single particle 3D reconstructions to high (<3.5Å) resolution. Emma’s role now is as a cryo-EM Support Scientist in the Astbury Biostructure Laboratory and is involved in a broad range of research questions all using high resolution electron microscopy.
Dr Matthew Iadanza

University of Leeds
Matt completed his Ph.D. in the lab of Dr. Tamir Gonen at the University of Washington in Seattle, where he worked developing microcrystal electron diffraction (microED). He is currently a senior research fellow in the labs of Sheena Radford and Neil Ranson at the University of Leeds, where he uses single-particle and helical reconstruction cryoEM techniques to study the structure of amyloid fibrils and the folding and insertion of proteins into the outer membrane of Gram-negative bacteria.
Rachel Johnson

University of Leeds
Rachel is a final year PhD student at the University of Leeds working under the supervision of Stephen Muench and Colin Fishwick. Her PhD project uses cryo-EM to aid structure-based drug design and focuses on determining the structure of membrane proteins which could act as anti-parasitic therapeutic targets
Mr David Kleb

University of Leeds
David is a student on the 4-year Wellcome trust PhD programme 'The Molecular Basis of Biological Mechanisms' at the Astbury Centre, University of Leeds. He is supervised by Stephen Muench and Frank Sobott and currently in his second year. His project is focused on developing a rapid grid preparation technique for cryo-EM which he aims to apply to time resolved structural studies.
Dr Dimitrios Kontziampasis

University of Leeds
Dr Dimitrios Kontziampasis is currently working in the Astbury Centre for Structural Molecular Biology, on the development of methodologies that can trap conformational states in the μs-ms timeframe for time resolved single particle cryo-Electron Microscopy. His main interests revolve around inter/crossdisciplinary and applied research, with a focus on the surface modification of materials for biological, energy and several other applications, and their characterisation.
Dr Dan Maskell

University of Leeds
I completed my PhD at UCL working on biochemical and structural characterisation of protein complexes involved in kinetochore formation. I continued my research interests with postdoctoral studies into structural and functional characterisation of nucleoprotein complexes involved in retroviral integration using single particle EM and other biochemical techniques. I took another postdoctoral position at Leeds Where I worked on structural characterisation of viral genomes and their association with capsid proteins. Within Leeds I changed roles to become a support scientist at the Astbury Biostructure Laboratory, working with scientists to facilitate their use of EM to solve interesting biological problems.
Dr Stephen Muench

University of Leeds
Dr Stephen Muench is a lecturer in membrane proteins at the Univeristy of Leeds. Having trained in X-ray crystallography, he moved to the field of electron microscopy in 2006. His main interests are in using electron microscopy, primarily single particle cryo-EM, to resolve the structure and mechanism of membrane proteins and to use this to underpin inhibitor design
Dr Colin Palmer

Science and Technology Facilities Council
Colin Palmer is a structural biologist at STFC, working on CCP-EM, the Collaborative Computational Project for Electron cryo-Microscopy. After a PhD on electron tomography and more recent work in software engineering, his current role combines both these interests by focussing on computational methods for cryo-EM
Dr Neil Ranson

University of Leeds
Dr Christos Savva

University of Leicester
I am the facility manager at the Midlands Regional Cryo-EM Facility based at the University of Leicester. Prior to this I was a research assistant at the MRC Laboratory of Molecular Biology in Cambridge and an associate research scientist at Texas A&M University Microscopy and Imaging Center. I am a structural biologist with interest in the structure-function relationships of pore-forming proteins made by micro-organisms and bacteriophages.
Dr Charlotte Scarff

University of Leeds
Dr Scarff is a Wellcome Trust ISSF Early Career Research Fellow at the University of Leeds. Her research interests lie in combining mass spectrometry and cryo-electron microscopy approaches to understand heterogeneity in macromolecular systems.

Further Information and Venue

This course will take place at the University of Leeds.

Registration fees and accommodation

Registration fees for this 4 day course are as follows:

RMS Member rate £1100
Non-member rate £1200

Accommodation package £264

Accommodation can be booked during the registration process. Accommodation has been reserved at the Ibis Hotel Leeds Centre Marlborough Street.
The accommodation package includes 4 nights bed and breakfast accommodation and transportation between the hotel and the course each day.

Single Particle Cryo-TEM School

Single Particle Cryo-TEM School

Outline Schedule

Day1

Day 2-4

Day 4

Organiser and Speaker List

Organiser

Dr Rebecca Thompson

Confirmed Speakers

Mr Alan Boswell

Dr Dan Clare

Dr Juan Fontana

Dr Emma Hesketh

Dr Matthew Iadanza

Rachel Johnson

Mr David Kleb

Dr Dimitrios Kontziampasis

Dr Dan Maskell

Dr Stephen Muench

Dr Colin Palmer

Dr Neil Ranson

Dr Christos Savva

Dr Charlotte Scarff

Further Information and Venue

Registration fees and accommodation

Sponsors

Agar Scientific

Gatan

Leica Microsystems

Thermo Fisher Scientific

TTP Labtech