Determination Our state-of-the-art cryo-electron microscopy (cryo-EM) service offers a seamless, fully integrated workflow for protein 3D structure determination, combining advanced wet lab techniques with high-performance computing infrastructure. Operating from a unified, fully commercial facility, we ensure efficient and thorough processing of samples through a meticulously designed three-phase process.
Phase 1: Pilot Study The process begins with a pilot study to assess the feasibility of target proteins, aiming to resolve 2D particle classes from a smaller cryo-EM dataset. This initial phase is crucial for laying the groundwork for more detailed structure analysis and aligning on the ideal strategy for achieving a 3D structure.
Phase 2: Optimization After a successful pilot study, we proceed to the optimization phase, focusing on cryo-grid refinement to enhance particle concentration, orientation, and other critical parameters. Our methods includes size exclusion chromatography, gradient density ultracentrifugation, membrane protein reconstitution complemented by additional cryo-EM screenings to fine-tune conditions for optimal data acquisition.
Phase 3: 3D Structure Determination The final phase is dedicated to the comprehensive determination of 3D structure. With data collected from optimized grids, we conduct extensive 2D and 3D classifications of particles, culminating in the reconstruction of a detailed 3D density map. Where necessary, additional modeling is performed by our experts.
Project Management and Timelines Each project begins with a meeting to outline objectives, timelines, and key stakeholders. Regular, specific alignment meetings ensure all parties are kept informed of progress and intermediate results. Timelines are tailored to each project, with the pilot study phase lasting 2-3 weeks, sample optimization approximately 2 weeks per iteration, and the 3D structure determination phase, depending on the target’s complexity, taking between 4 to 8 weeks. Our integrated approach, from lab to computational analysis, guarantees precise, high-quality results for our clients, pushing the boundaries of structural biology research.
We prepare everything else. We receive your sample, then optimise and test it before returning the data in a personalised way to meet your needs.
