Graphene Oxide-Supported Microwell Grids for Preparing Cryo-EM Samples with Controlled Ice Thickness.

Autor: Kang MH; School of Chemical and Biological Engineering, and Institute of Chemical Processes (ICP), Seoul National University, Seoul, 08826, Republic of Korea.; Center for Nanoparticle Research, Institute of Basic Science (IBS), Seoul, 08826, Republic of Korea., Park J; School of Biological Sciences, Seoul National University, Seoul, 08826, Republic of Korea., Kang S; School of Chemical and Biological Engineering, and Institute of Chemical Processes (ICP), Seoul National University, Seoul, 08826, Republic of Korea.; Center for Nanoparticle Research, Institute of Basic Science (IBS), Seoul, 08826, Republic of Korea., Jeon S; Department of Mechanical Engineering, BK21FOUR ERICA-ACE Center, Hanyang University, Ansan, Gyeonggi, 15588, Republic of Korea., Lee M; School of Chemical and Biological Engineering, and Institute of Chemical Processes (ICP), Seoul National University, Seoul, 08826, Republic of Korea.; Center for Nanoparticle Research, Institute of Basic Science (IBS), Seoul, 08826, Republic of Korea., Shim JY; Department of Chemical and Biological Engineering, Sookmyung Women's University, Seoul, 04310, Republic of Korea., Lee J; Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Jeon TJ; National Instrumentation Center for Environmental Management, Seoul National University, Seoul, 08826, Republic of Korea., Ahn MK; Department of Materials Science and Engineering, Seoul National University, Seoul, 08826, Republic of Korea.; Biomedical Implant Convergence Research Lab, Advanced Institutes of Convergence Technology, Suwon, 16229, Republic of Korea., Lee SM; Department of Materials Science and Engineering, Seoul National University, Seoul, 08826, Republic of Korea.; Biomedical Implant Convergence Research Lab, Advanced Institutes of Convergence Technology, Suwon, 16229, Republic of Korea., Kwon O; National Instrumentation Center for Environmental Management, Seoul National University, Seoul, 08826, Republic of Korea., Kim BH; Department of Organic Materials and Fiber Engineering, Soongsil University, Seoul, 06978, Republic of Korea., Meyerson JR; Department of Physiology and Biophysics, Weill Cornell Medical College of Cornell University, New York, NY, 10065, USA., Lee MJ; Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Lim KI; Department of Chemical and Biological Engineering, Sookmyung Women's University, Seoul, 04310, Republic of Korea., Roh SH; School of Biological Sciences, Seoul National University, Seoul, 08826, Republic of Korea., Lee WC; Department of Mechanical Engineering, BK21FOUR ERICA-ACE Center, Hanyang University, Ansan, Gyeonggi, 15588, Republic of Korea., Park J; School of Chemical and Biological Engineering, and Institute of Chemical Processes (ICP), Seoul National University, Seoul, 08826, Republic of Korea.; Center for Nanoparticle Research, Institute of Basic Science (IBS), Seoul, 08826, Republic of Korea.
Jazyk: angličtina
Zdroj: Advanced materials (Deerfield Beach, Fla.) [Adv Mater] 2021 Oct; Vol. 33 (43), pp. e2102991. Date of Electronic Publication: 2021 Sep 12.
DOI: 10.1002/adma.202102991
Abstrakt: Cryogenic-electron microscopy (cryo-EM) is the preferred method to determine 3D structures of proteins and to study diverse material systems that intrinsically have radiation or air sensitivity. Current cryo-EM sample preparation methods provide limited control over the sample quality, which limits the efficiency and high throughput of 3D structure analysis. This is partly because it is difficult to control the thickness of the vitreous ice that embeds specimens, in the range of nanoscale, depending on the size and type of materials of interest. Thus, there is a need for fine regulation of the thickness of vitreous ice to deliver consistent high signal-to-noise ratios for low-contrast biological specimens. Herein, an advanced silicon-chip-based device is developed which has a regular array of micropatterned holes with a graphene oxide (GO) window on freestanding silicon nitride (Si x N y ). Accurately regulated depths of micropatterned holes enable precise control of vitreous ice thickness. Furthermore, GO window with affinity for biomolecules can facilitate concentration of the sample molecules at a higher level. Incorporation of micropatterned chips with a GO window enhances cryo-EM imaging for various nanoscale biological samples including human immunodeficiency viral particles, groEL tetradecamers, apoferritin octahedral, aldolase homotetramer complexes, and tau filaments, as well as inorganic materials.
(© 2021 Wiley-VCH GmbH.)
Databáze: MEDLINE
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