Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

The mechanism of ribosomal recruitment during translation initiation on the Type 2 encephalomyocarditis virus IRES.

The EMBO journal·2026

Same author

Structural Studies of Nedicistrovirus IRES-Driven, Initiation Factor-independent Translation Shed Light on Key Steps of Eukaryotic Translation Elongation.

bioRxiv : the preprint server for biology·2025

Same author

Inconsistencies in the published rabbit ribosomal rRNAs: a proposal for uniformity in sequence and site numbering.

RNA (New York, N.Y.)·2025

Same author

Passage through micro-sprayer increases functional activity - implications for activity assays in time-resolved cryo-EM.

bioRxiv : the preprint server for biology·2024

Same author

Inconsistencies in the published rabbit ribosomal rRNAs: a proposal for uniformity in sequence and site numbering.

bioRxiv : the preprint server for biology·2024

Same author

Recovery of Conformational Continuum From Single-Particle Cryo-EM Images: Optimization of ManifoldEM Informed by Ground Truth.

IEEE transactions on computational imaging·2022

Search research articles

Related Experiment Video

Updated: Jun 22, 2026

A Microfluidic Chip for ICPMS Sample Introduction

A Microfluidic Chip for ICPMS Sample Introduction

Published on: March 5, 2015

Time-resolved cryo-EM (TRCEM) sample preparation using a PDMS-based microfluidic chip assembly.

Xiangsong Feng¹, Joachim Frank^1,2

¹Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY 10032.

Biorxiv : the Preprint Server for Biology

|December 23, 2024

Summary

This study introduces a novel time-resolved cryo-electron microscopy (TRCEM) setup using microfluidics for rapid sample preparation. This method provides crucial structural and kinetic insights into biomolecular reactions within milliseconds.

Keywords:

Micro/nanofabrication Microfluidics Single-particle cryo-EM Time-resolved cryo-EM sample preparation

More Related Videos

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Published on: November 6, 2021

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton

Published on: February 25, 2021

Related Experiment Videos

Last Updated: Jun 22, 2026

A Microfluidic Chip for ICPMS Sample Introduction

A Microfluidic Chip for ICPMS Sample Introduction

Published on: March 5, 2015

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Published on: November 6, 2021

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton

Published on: February 25, 2021

Area of Science:

Structural Biology
Biochemistry
Cryo-Electron Microscopy

Background:

Traditional cryo-electron microscopy (cryo-EM) methods are limited in capturing transient states of biomolecular reactions.
Existing time-resolved cryo-EM (TRCEM) techniques face challenges in achieving millisecond time resolution and efficient sample preparation.
Understanding pre-equilibrium intermediates is crucial for elucidating molecular mechanisms.

Purpose of the Study:

To develop and present a novel TRCEM setup for capturing biomolecular reactions at millisecond timescales.
To overcome limitations of sample adsorption and ineffective mixing in previous TRCEM methods.
To demonstrate the application of the developed TRCEM method in studying ribosome recycling.

Main Methods:

Development of a microfluidics chip assembly using polydimethylsiloxane (PDMS).
The assembly includes an SiO2-coated micromixer, a glass-capillary microreactor, and a microsprayer for sample deposition.
Application of the TRCEM setup to investigate the mechanism of ribosome recycling mediated by High frequency of lysogenization X (HflX).

Main Results:

The novel TRCEM setup effectively addresses issues of sample adsorption and fluid mixing.
The method yields highly reproducible results, demonstrated in the study of translation.
Investigation of HflX-mediated ribosome recycling provided biologically significant, reproducible structural and kinetic information.

Conclusions:

The presented microfluidics-based TRCEM setup enables structural and kinetic analysis of biomolecular reactions in the 10-1000 ms time range.
This protocol offers a promising approach for studying pre-equilibrium intermediates in diverse biological systems.
The developed TRCEM device is effective for obtaining high-quality data for mechanistic studies.