Simplified approach for preparing graphene oxide tem grids for stained and vitrified biomolecules

Kumar, A and Sengupta, N and Dutta, S (2021) Simplified approach for preparing graphene oxide tem grids for stained and vitrified biomolecules. In: Nanomaterials, 11 (3). pp. 1-22.

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Abstract

In this manuscript, we report the application of graphene oxide (GO) in the preparation of cryo�electron microscopy (cryo�EM) and transmission electron microscopy (TEM) grids. We treated GO with water and organic solvents, such as, methanol, ethanol and isopropanol separately to isolate significantly large GO monolayer flake to fabricate the grids for cryo�EM and TEM study. We implemented a simplified approach to isolate flakes of GO monolayer for constructing the TEM grids, independent of expensive heavy equipment (Langmuir�Blodgett trough, glow�discharge system, carbon�evaporator or plasma�cleaner or peristaltic pumps). We employed confocal microscopy, SEM and TEM to characterize the flake size, stability and transparency of the GO monolayer and atomic force microscopy (AFM) to probe the depth of GO coated grids. Additionally, GO grids are visualized at cryogenic condition for suitability of GO monolayer for cryo�EM study. In addition, GO�Met�H2O grids reduce the effect of preferred orientation of biological macromolecules within the amorphous ice. The power�spectrum and contrast�transfer�function unequivocally suggest that GO�Met�H2O fabricated holey grids have excellent potential for application in high�resolution structural characterization of biomolecules. Furthermore, only 200 movies and ~8000 70S ribosome particles are selected on GO�coated grids for cryo�EM reconstruction to achieve high�resolution structure. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Item Type:	Journal Article
Publication:	Nanomaterials
Publisher:	MDPI AG
Additional Information:	The copyright for this article belongs to MDPI AG
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	24 Mar 2021 11:26
Last Modified:	24 Mar 2021 11:26
URI:	http://eprints.iisc.ac.in/id/eprint/68513

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