Introducing a novel time-resolved cryo-EM method by the Frank Lab at Columbia University

The CryoSol Community welcomes you to share knowledge about sample preparation for cryo-electron microscopy (cryo-EM) and learn from each other. We believe this is vital to advancing cryo-EM research. Our Community is open to presenting publications, new insights, and practical knowledge to innovate cryo-EM sample preparation together.

We are delighted to extend a warm welcome to Professor Joachim Frank and Dr. Xiangsong Feng from the Frank Lab at Columbia University's Department of Biochemistry and Molecular Biophysics as our distinguished guest speakers for the upcoming Cryo-EM Community webinar.

Professor Joachim Frank is a pioneering biophysicist whose transformative contributions to single-particle cryo-electron microscopy (cryo-EM) have revolutionized our ability to visualize biological molecules in exquisite detail. His groundbreaking work in this field earned him the Nobel Prize in Chemistry in 2017, shared with Jacques Dubochet and Richard Henderson. Professor Frank's extensive research has profoundly deepened our understanding of the structural intricacies and functional dynamics of ribosomes in both bacteria and eukaryotes, earning him widespread recognition and numerous accolades in the scientific community.

Joining him is Dr. Xiangsong Feng, a postdoctoral research scientist at Columbia University Medical Center, whose expertise lies at the intersection of time-resolved cryo-electron microscopy and microfluidics. Dr. Feng is renowned for his innovative work in fluid mechanics, microfabrication, and passive micromixing mechanisms. He has authored several influential publications in these areas, contributing significantly to the advancement of cryo-EM techniques.

In their presentation, "Time Resolution in Cryo-EM Using a Novel PDMS-Based Microfluidic Chip Assembly and Its Application to the Study of HflX-Mediated Ribosome Recycling," Professor Frank and Dr. Feng will share insights from their latest research, showcasing how their novel approach enhances the temporal resolution in cryo-EM studies. We eagerly anticipate their discussion and the profound implications it holds for the future of structural biology.

Register now to save your seat for this webinar:

Introducing a novel time-resolved cryo-EM method by the Frank lab at Columbia University

Tuesday July 9th at 5 PM CET | 11 AM EST | 8 AM PST

ABSTRACT OF THE RECENT CELL PUBLICATION (Cell.com)

"Time resolution in cryo-EM using a PDMS-based microfluidic chip assembly and its application to the study of HflX-mediated ribosome recycling’’

The rapid kinetics of biological processes and associated short-lived conformational changes pose a significant challenge in attempts to structurally visualize biomolecules during a reaction in real time. Conventionally, on-pathway intermediates have been trapped using chemical modifications or reduced temperature, giving limited insights. Here, we introduce a time-resolved cryo-EM method using a reusable PDMS-based microfluidic chip assembly with high reactant mixing efficiency. Coating of PDMS walls with SiO2 virtually eliminates non-specific sample adsorption and ensures maintenance of the stoichiometry of the reaction, rendering it highly reproducible. In an operating range from 10 to 1,000 ms, the device allows us to follow in vitro reactions of biological molecules at resolution levels in the range of 3 Å. By employing this method, we show the mechanism of progressive HflX-mediated splitting of the 70S E. coli ribosome in the presence of the GTP via capture of three high-resolution reaction intermediates within 140 ms.

AN INTRODUCTION TO THEIR SCIENTIFIC JOURNEY IN CRYO-EM AND MICROFLUIDIC CHIP DEVELOPMENT

The research presented in this publication spans over a decade, focusing on groundbreaking advancements in cryo-electron microscopy (cryo-EM) and microfluidic technologies to explore complex biological processes. This journey began before 2008 at the Wadsworth Center in Albany, New York, under the leadership of Terry Wagenknecht, where the team embarked on pioneering time-resolved cryo-EM studies. During this period, Zonghuan Lu, a postdoctoral researcher at Rensselaer Polytechnic Institute, made significant contributions by developing a novel monolithic silicon-based microfluidic chip for mixing, reacting, and spraying. This innovation, crucial for ribosome-related experiments, was published in J. Struct. Biol. in 2009.

In 2008, upon transitioning to Columbia University, the team, with the assistance of graduate student Bo Chen and Columbia's Engineering Department, adapted these silicon chips for time-resolved experiments. They utilized a computer-operated system developed by Howard White from Leeds, England, which facilitated precise control over the experiments.

The research evolved further with Xiangsong Feng's efforts under the co-mentorship of Dr. Qiao Lin. Xiangsong's work led to the development of a PDMS-based chip, which was highlighted in their recent CELL publication. This advancement allowed for more refined and dynamic studies of biological processes. Xiangsong's collaboration with the group, from his Ph.D. studies through his postdoctoral work, has been critical in advancing the capabilities of microfluidic applications in cryo-EM.

https://joachimfranklab.org/the-team/

LIVE Q&A

The webinar will be hosted by René Henderikx, the VitroJet Product Manager at CryoSol-World, who is responsible for the roadmap of sample preparation solutions for cryo-EM. Co-hosting the webinar will be Cliff Mathisen from Nanoscience Instruments, the authorized VitroJet distributor in North America. CryoSol’s CTO, Bart Beulen, will also be present. They will join the live Q&A session with our guest speakers following their live presentation.

We encourage you to ask many questions during the presentation to inspire and support your needs in cryo-EM and sample preparation. We believe that by working together, we can achieve more: two achieve more.

You will receive an email instantly after registration, including a link to access the webinar. Enjoy the webinar and in case of questions do not hesitate to reach out to info@cryosol-world.com.