BPS2026 – Adapting contactless ECM micropatterning for high-resolution cryo-EM studies of cell-cell junctions and their molecular machinery

This project aims to investigate molecular interactions between cells in their native, hydrated state In particular, we want to study how force and tension (mechanotransduction) affect cell-cell adherens junctions in mammalian kidney epithelial cells, which are composed of proteins such as cadherin, catenin, and actin At present, cryogenic transmission electron microscopy (cryo-TEM) and cryogenic electron tomography (cryo-ET)—rapidly advancing fields in structural biology—are the only methods that allow researchers to study these molecular machines in fully hydrated, frozen cells in 3D at near-atomic resolution To address challenges with reproducibility in imaging cell-cell junctions, we are optimizing and streamlining a system developed in our lab for treating substrates with maskless extracellular matrix protein micropatterning This approach controls cell attachment, position, and shape, making it possible to produce cell doublets with clear cell-cell contacts on TEM-compatible substrates Our goal is to measure the forces generated by these micropatterns and determine their impact on junctional reinforcement This work will advance understanding of cell-cell interactions that play key roles in processes such as tissue development and disease progression Beyond this specific system, a completed and validated generalized workflow will serve as an invaluable resource for high-resolution cryo-TEM studies of intact hydrated cells, with a particular focus on mechanotransduction.