Amino and PEG-amino graphene oxide grids enrich and protect samples for high-resolution single particle cryo-electron microscopy |
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| Affiliation: | 1. Department of Biochemistry & Biophysics and the Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94143, United States;2. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, United States;1. Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University, Leninskie Gory 1, Bld 12, Moscow 119991, Russia;2. Belozersky Research Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119234, Russia;3. National Research Center “Kurchatov Institute”, Moscow 123098, Russia;4. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia;1. Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA;2. Department of Chemistry, University of California, Berkeley, CA 94720, USA;3. Department of Molecular and Cell Biology and California Institute of Quantitative Biosciences, University of California, Berkeley, CA 94720, USA;1. Department of Cell Biology, UT Southwestern Medical Center, 6000 Harry Hines Blvd, Dallas, TX 75390, USA;2. Department of Biochemistry, UT Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390, USA;3. Department of Biophysics, UT Southwestern Medical Center, 6000 Harry Hines Blvd, Dallas, TX 75390, USA;1. The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, 89 Convent Ave, New York, NY 10027, USA;2. Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA;1. Department of Biological Science, Markey Center for Structural Biology, Purdue University, West Lafayette, IN 47907, USA;2. Divisions of Infectious Diseases, Cincinnati Children''s Hospital Medical Center, Cincinnati, OH 45229, USA;3. Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA |
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| Abstract: | Cryo-EM samples prepared using traditional methods often suffer from too few particles, poor particle distribution, strongly biased orientation, or damage from the air-water interface. Here we report that functionalization of graphene oxide (GO) coated grids with amino groups concentrates samples on the grid with improved distribution and orientation. By introducing a PEG spacer, particles are kept away from both the GO surface and the air-water interface, protecting them from potential denaturation. |
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| Keywords: | Cryo-EM Graphene oxide Sample preparation Amino functionalization Air-water interface |
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