Investigating Ultra-High-Quality Graphene-Based Cryo-EM Device Architecture for Higher Reproducibility, Speed, and Resolution

Project Summary Cryogenic Electron Microscopy (Cryo-EM) has revolutionized macromolecule imaging by providing near-atomic scale resolution insights into complex biological molecules' structures. However, current sample preparation methods face challenges like denaturation, aggregation, and preferential alignment, limiting reproducibility and efficiency. The limitations of imaging methods hinder scientific advancements which directly affect the ability to develop novel vaccines and drug discovery. As AI-guided computational techniques emerge for mapping protein structures, there is a need for imaging methods to evolve alongside. Graphene offers an ideal solution for addressing these challenges. Sindri Materials has developed unique, ultra-high-quality (UHQ), large-area graphene production that is reproducibly atomically clean and continuous. This graphene is critical for observing unobscured atomic features and ensuring survivability during cryo-EM sample preparation. Preliminary results strongly suggest that graphene growth quality is more intricately connected with high-quality transfer than previously thought, necessitating a renewed investigation into transfer methodologies. The goal is to develop a graphene-based cryo-EM device architecture that preserves key UHQ characteristics and enables reliable, reproducible, and rapid cryo-EM integration. This will include investigating factors such as surface adhesion, transfer-induced defects, and plasma treatment in transferring to produce a clean, continuous, and strong graphene grid suitable for cryo-EM. Aim 1 examines the development of an optimized graphene transfer method by examining support materials, quantifying transfer-induced residue, and assessing the mechanical response of graphene. The aim is to maximize graphene integrity and minimize sample damage during the process of transferring graphene onto a cryo-EM grid. Aim 2 examines the effect of varying degrees of plasma treatment on the mechanical performance of graphene, an important factor in cryo-EM sample preparation. The aim is to determine the extent of plasma exposure before significant mechanical property deterioration occurs. This research aims to establish the methodology to fabricate UHQ graphene-based cryo-EM device architecture necessary to address rampant image capture challenges faced by structural biologists, leading to higher reproducibility, speed, and resolution, and therefore expanding the capabilities of researchers to develop groundbreaking advancements in therapeutics and vaccines improving human health. The success of this research will demonstrate the technical feasibility and define the parameters necessary for scaling up commercial production to meet the growing market demand in a potential Phase II.