Life in slow motion: smart multimodal imaging at kilohertz rates

PROJECT SUMMARY This project aims to develop new scalable nD cellular and unicellular organism imaging and analysis approaches to study the ‘rules of life.’ We propose to develop a novel multimodal imaging platform, blending fluorescence super-resolution and tomographic polarized light imaging. Paired with the proposed platform, we will provide an open-source suite of computational imaging tools to recover the dielectric tensor in multiple scattering samples using GPU acceleration. As part of these tools, we address optimal pattern generation, correct for phase drift, test against rigorous simulations, and integrate recent advancement in deep learning denoisers. We will benchmark the platform’s performance on two cellular motility projects. In the first project (Aim 2), we will quantify both 3D hydrodynamic flow fields surrounding motile bacteria using tomographic polarized light imaging and molecular cell fate markers using fluorescence. We then aim to demonstrate two unique measurements by quantifying the flow field and cell fate of motile bacteria near or at surfaces and near the edge of a swarming bacteria colony. In the second project (Aim 3), we will quantify for the first time the local 3D viscoelastic properties and applied forces for eukaryotic cells placed into collagen matrices. We will validate our measurements using known atomic force microscopy and standard 2D traction force measurements before attempting to generate self-calibrated 3D traction force microscopy data by quantifying the local viscoelastic environment surrounding the motile cells. Across all three aims, we intend to demonstrate a multimodal imaging platform that is easily adaptable and addresses critical needs as the community moves towards probing biology at the fastest rates possible.