Video-rate quantitative phase and polarimetric imaging (QP2I) for label-free imaging of contracting muscle cells

Abstract

Quantitative phase imaging (QPI) has become a standard approach for label-free microscopy of transparent specimens, mapping the optical path length. However, the isotropic phase is blind to the molecular order underlying optical anisotropy. Implementations that also add quantitative contrast to birefringence rely on multi-frame illumination or polarization-state scanning, or on dedicated interferometric hardware and thus trade away acquisition speed or the simplicity of a standard microscope. We present Quantitative Phase and Polarimetric Imaging (QPPI), a computational imaging framework that converts a standard differential interference contrast (DIC) microscope with a polarization camera into a single-shot, video-rate, dual-contrast imaging system. We developed a new globally regularized phase reconstruction that eliminates artifacts of prior DIC-to-QPI approaches, providing quantitative phase information, and used per-pixel Stokes-based reconstruction to obtain quantitative anisotropic retardance. We validate the approach across simulations, static reference targets, fixed muscle tissue and living dynamically moving cells. QPPI enables all-optical and label-free studies of contractile dynamics in live cardiomyocytes. It provides both structural and functional readouts at sufficient frame rates to resolve fast cardiomyocyte contractions. Conventional fluorescent labels bleach over time, perturb physiology, and preclude long-term monitoring. Instead, QPPI enables long-term, label-free interrogation of dynamic cellular processes at quantitative contrast and provides structural readouts similar to those provided by established fluorescent labels, such as frame-by-frame sarcomere length. It also offers previously inaccessible measurements, like quantitative analysis of retardance changes during muscle contraction, which provides a direct optical signature of muscle photoelasticity.