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

ABSTRACT: Time-resolved cryo-EM (TRCEM) makes it possible to provide structural and kinetic information on a reaction of biomolecules before the equilibrium is reached. Several TRCEM methods have been developed in the past to obtain key insights into the mechanism of action of molecules and molecular machines on the time scale of tens to hundreds of milliseconds, which is unattainable by the normal blotting method. Here we present our TRCEM setup utilizing a polydimethylsiloxane (PDMS)-based microfluidics chip assembly, comprising three components: a PDMS-based, internally SiO₂-coated micromixer, a glass-capillary microreactor, and a PDMS-based microsprayer for depositing the reaction product onto the EM grid. As we have demonstrated in recent experiments, this setup is capable of address

SUBMITTER: Feng X 

PROVIDER: S-EPMC11661195 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature