한빛사논문
- 조회1,357
Min-Ho Kanga,b, Junsun Parkc, Sungsu Kanga,b, Sungho Jeond, Minyoung Leea,b, Ji-Yeon Shime, Jeeyoung Leef, Tae Jin Jeong, Min Kyung Ahnh,i, Sung Mi Leeh,i, Ohkyung Kwong, Byung Hyo Kimj, Joel R. Meyersonk, Min Jae Leef, Kwang-Il Lime,*, Soung-Hun Rohc,*, Won Chul Leed,* and Jungwon Parka,b,*
aSchool of Chemical and Biological Engineering, and Institute of Chemical Processes (ICP), Seoul National University, Seoul 08826, Republic of Korea
bCenter for Nanoparticle Research, Institute of Basic Science (IBS), Seoul 08826, Republic of Korea
cSchool of Biological Sciences, Seoul National University, Seoul 08826, Republic of Korea
dDepartment of Mechanical Engineering, BK21FOUR ERICA-ACE Center, Hanyang University, Ansan, Gyeonggi 15588, Republic of Korea
eDepartment of Chemical and Biological Engineering, Sookmyung Women’s University, Seoul 04310, Republic of Korea
fDepartment of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
gNational Instrumentation Center for Environmental Management, Seoul National University, Seoul 08826, Republic of Korea
hDepartment of Materials Science and Engineering, Seoul National University, Seoul 08826, Republic of Korea
iBiomedical Implant Convergence Research Lab, Advanced Institutes of Convergence Technology, Suwon 16229, Republic of Korea
jDepartment of Organic Materials and Fiber Engineering, Soongsil University, Seoul 06978, Republic of Korea
kDepartment of Physiology and Biophysics, Weill Cornell Medical College of Cornell University, New York, NY 10065, USA
M.-H.K. and J.P. contributed equally to this work.
*To whom correspondence should be addressed.
Abstract
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 (SixNy). 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.
논문정보
- Research article
- 2021년 09월 (BRIC 등록일 2021-10-01)
- 국내(교신)+국외 연구진
- 바이오·의료융합 > 바이오소재
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