Membrane proteins play important roles in biological functions, with accompanying allosteric structure changes. Understanding intramolecular dynamics helps elucidate catalytic mechanisms and develop new drugs. In contrast to the various technologies for structural analysis, methods for analyzing intramolecular dynamics are limited. Single-molecule measurements using optical microscopy have been widely used for kinetic analysis. Recently, improvements in detectors and image analysis technology have made it possible to use single-molecule determination methods using X-rays and electron beams, such as diffracted X-ray tracking (DXT), X-ray free electron laser (XFEL) imaging, and cryo-electron microscopy (cryo-EM). High-speed atomic force microscopy (HS-AFM) is a scanning probe microscope that can capture the structural dynamics of biomolecules in real time at the single-molecule level. Time-resolved techniques also facilitate an understanding of real-time intramolecular processes during chemical reactions. In this review, recent advances in membrane protein dynamics visualization techniques were presented.

Measuring molecular dynamics improves understanding of the structure–function relationships of materials. In this study, we present a novel technique for observing material dynamics using transmission electron microscopy (TEM), in which the gold nanoparticles are employed as motion probes for tracing the polymer dynamics in real space. A thin layer of polymer materials was generated on the 2 μm diameter holes of Quantifoil grids, and gold nanoparticles were dispersed on the membrane surface. By tracking the movement of gold nanoparticles from a series of TEM images taken under continuous temperature control, we obtained mean squared displacement (MSD) curves. The dynamics of poly{2-(perfluorooctyl)ethyl acrylate} (PC₈FA) and poly(stearyl acrylate) (PSA) were analyzed. In the temperature-dependent analysis of the MSD, sharp peaks were observed for both PC₈FA and PSA at positions corresponding to their melting and crystallization temperatures. These results demonstrate the capability of TEM to provide valuable insights into the dynamics of polymer materials, highlighting its potential for widespread application in materials sciences.