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Theoretical framework and experimental solution for the air-water interface adsorption problem in cryoEM

Biophysics Reports 2023年第4期9卷 215-229页

作者： Joon S.Kang Xueting Zhou Yun-Tao Liu Kaituo Wang Z.Hong Zhou Department of Microbiology Immunology & Molecular Genetics University of California Los Angeles (UCLA) Molecular Biology Institute UCLA California Nano Systems Institute UCLA

As cryogenic electron microscopy(cryoEM) gains traction in the structural biology community as a method of choice for determining atomic structures of biological complexes, it has been increasingly recognized that many complexes that behave well under conventional negative-stain electron microscopy tend to have preferential orientation, aggregate or simply mysteriously “disappear” on cryoEM grids. However, the reasons for such misbehavior are not well understood, which limits systematic approaches to solving the problem. Here, we have developed a theoretical formulation that explains these observations. Our formulation predicts that all particles migrate to the air–water interface(AWI)to lower the total potential surface energy-rationalizing the use of surfactant, which is a direct solution to reduce the surface tension of the aqueous solution. By performing cryogenic electron tomography(cryo ET) on the widely-tested sample, Gro EL, we demonstrate that, in a standard buffer solution, nearly all particles migrate to the AWI. Gradually reducing the surface tension by introducing surfactants decreased the percentage of particles exposed to the surface. By conducting single-particle cryoEM, we confirm that suitable surfactants do not damage the biological complex, thus suggesting that they might provide a practical, simple, and general solution to the problem for high-resolution cryoEM. Applying this solution to a real-world AWI adsorption problem involving a more challenging membrane protein,namely, the Cl C-1 channel, has resulted in its near-atomic structure determination using cryoEM.

关键词： CryoEM cryoet Sample preparation Air-water interface adsorption Surfactant Surface energy

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Ultrastructural analysis of neuronal synapses using state-of-the-art nano-imaging techniques

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Neuroscience Bulletin 2012年第4期28卷 321-332页

作者： Changlu Tao Chenglong Xia Xiaobing Chen Z. Hong Zhou Guoqiang Bi Center for Integrative Imaging Hefei National Laboratory for Physical Sciences at the Microscale & School of LifeSciences University of Science and Technology of China Hefei 230027 China Laboratory of Neurobiology National Institute of Neurological Disorders and Stroke National Institutes of HealthBethesda 20892 USA Department of Microbiology Immunology & Molecular Genetics University of California Los Angeles Los Angeles CA90095-7364 USA

Neuronal synapses are functional nodes in neural *** organization and activity define an individual＇s level of intelligence,emotional state and mental *** in the structure and efficacy of synapses are the biological basis of learning and ***,investigation of the molecular architecture of synapses has been impeded by the lack of efficient techniques with sufficient *** developments in state-of-the-art nano-imaging techniques have opened up a new window for dissecting the molecular organization of neuronal synapses with unprecedented ***,we review recent technological advances in nano-imaging techniques as well as their applications to the study of synapses,emphasizing super-resolution light microscopy and 3-dimensional electron tomography.

关键词： synaptic architecture nano-imaging super-resolution imaging STED microscopy STORM PALM cryoet

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