On Demand CryoET Mini Symposium: Sample Preparation – The major limitation for CryoET and SPA

July 14, 2021 | News Type: Technology:

Have you been curious about the newest advancements in TEM sample preparation technology?

Both single particle analysis (SPA) and Cryo electron tomography (cryoET) workflows have grown exponentially in their popularity and usage in structural biology research. Unfortunately, both techniques are largely limited in their success due to high rates of sample preparation-related failures. Considering the increasing demand for CryoEM/ET data along with the enormous costs associated with operating cryoTEM facilities, overcoming this problem is imperative.

Please join us for our upcoming mini symposium where Dr. Peter J. Peters (Maastricht University) and Dr. Stefan Raunser (Max Planck Institute) will discuss advances in sample preparation instrumentation for CryoET and SPA, and their exciting new research that has been enabled by these technologies. Giulia Weissenberger from CryoSol will also provide an update on the first successful VitroJet installation in the world, completed in May 2021 at Oregon Health and Science University.


Webinar Details

Originally aired: June 22nd, 2021 at 1pm EDT

Presenters

Peter J. Peters, PhD

Distinguished University Professor, Limburg Chair, and Co-Director of the Maastricht MultiModal Molecular Imaging Institute (M4I), Maastricht University

Dr. Peters will discuss the technology behind the Vitrojet that he and his team at Maastricht University have been instrumental in pioneering. The Vitrojet encompasses ethane jetting, sample printing, robotic handling, dew point control, and pre-clipping of autogrids for an 8-9x improvement in grid quality compared to plunge freezing.

Stefan Raunser, PhD

Director, Department of Structural Biochemistry – Max Planck Institute of Molecular Physiology,
Technical University Dortmund

Dr. Raunser will discuss the new cryoET ice contamination reduction technology that he helped develop at the Max Planck Institute of Molecular Physiology. The Ceres Ice Defense system has demonstrated a reduction of nearly 100% of amorphous ice and >50% of crystalline ice that typically accumulates during sample prep.

VitroJet Installation Update

Giulia Weissenberger CryoSol-World

Giulia Weissenberger

Applications Scientist
CryoSol-World

Giulia will discuss the technology behind the VitroJet cryoEM sample preparation system, and the benefits of this innovative technique. She will also provide an update on the first successful installation in the world which enabled researchers at Oregon Health and Science University to create cryoEM samples with repeatable ice thickness and quality.

Moderator

Cliff Mathisen

Sales and Applications for TEM Solutions
Nanoscience Instruments


Abstracts

Understanding the invisible hands of sample preparation for cryo-EM

Presenter: Peter J. Peters, PhD
Director of The Maastricht Multimodal Molecular Imaging Institute (M4I)
Distinguished University Professor and Limburg Chair, Maastricht University
Maastricht, The Netherlands

Abstract:

Cryo-electron microscopy (cryo-EM) is rapidly becoming an attractive method in the field of structural biology. With the exploding popularity of cryo-EM, sample preparation must evolve to prevent congestion in the workflow. The dire need for improved microscopy samples has led to a diversification of methods. This talk aims to categorize and explain the principles behind various techniques in the preparation of vitrified samples for the electron microscope and will focus on the benefits and beauty of the Vitrojet. Various aspects and challenges in the workflow are discussed, from sample optimization and carriers to deposition and vitrification. Reliable and versatile specimen preparation remains a challenge, and we hope to give guidelines and posit future directions for improvement. We are also eager to hear your suggestions for collaborations in order to improve the technology. 

A streamlined cryo-ET workflow to obtain sarcomere structures at molecular resolution

Presenter: Stefan Raunser, PhD
Director and Scientific Member, Department of Structural Biochemistry –
Max Planck Institute of Molecular Physiology at Technical University Dortmund
Dortmund, Germany

Abstract:

Cryo-electron tomography (cryo-ET) is an emerging technique for studying cellular architecture and the structure of proteins at high resolution in situ. In this presentation, I will present our latest hardware developments in structural cell biology using FIB milling and cryo-ET. I will then explain how we used this workflow to obtain high-resolution structures of native sarcomeres, the force-generating, and load-bearing devices of muscles. Our cryo-ET reconstructions reveal molecular details of the three-dimensional organization and interaction of actin and myosin in the A-band, I-band and Z-disc and demonstrate that α-actinin cross-links antiparallel actin filaments by forming doublets with 6 nm spacing. Structures of myosin, tropomyosin, and actin at ~10 Å further reveal two conformations of “double-headed” myosin, where the flexible orientation of the lever arm and light chains enable myosin not only to interact with the same actin filament, but also to split between two actin filaments. Our results provide unexpected insights into the fundamental organization of vertebrate skeletal muscle and serve as a strong foundation for future investigations of muscle diseases.


Missed the live presentation?

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