2024/12/05 by Wei-Feng Xue, Wei‐Feng Xue
Biochemistry, Genetics and Molecular Biology · Engineering · Physics and Astronomy · #Advanced Electron Microscopy Techniques and Applications #Force Microscopy Techniques and Applications #Molecular Junctions and Nanostructures
paper · doi:10.1016/j.str.2024.11.007
openalex publication_date 2024/12/05 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/29
Atomic force microscopy (AFM) is a powerful and increasingly accessible technology that has a wide range of bio-imaging applications. AFM is capable of producing detailed three-dimensional topographical images with high signal-to-noise ratio, which enables the structural features of individual molecules to be studied without the need for ensemble averaging. Here, a software tool Tracey, designed to reconstruct the three-dimensional surface envelopes of individual helical filament structures from topographical AFM images, is presented. Workflow using Tracey is demonstrated on the structural analysis of individual helical amyloid protein fibrils where the assembly mechanism of heterogeneous, complex and diverse fibril populations due to structural polymorphism is not understood. The algorithms presented here allow structural information encoded in topographical AFM height images to be extracted and understood as three-dimensional (3D) contact point clouds. This approach will facilitate the use of AFM in structural biology to understand molecular structures and behaviors at individual molecule level.