Optical Volume Measurement of Beating Cardiac Myocytes using Quantitative Phase Imaging

Abstract

This paper describes recent research in developing a high-resolution optical microscope capable of real-time quantitative phase imaging to study morphological changes such as relative optical volume and optical thickness in cellular structures. A pixelated wire grid polarizer mask bonded to a CCD camera sensor enables simultaneous measurement of multiple phase contrast interference patterns. These data are processed in parallel to obtain phase, optical path difference (OPD), optical thickness (OT), optical volume (OV), simulated DIC (gradient), simulated dark field (gradient magnitude), and 3D typographic movies.Results will be presented of quantitative measurements of optical volume changes in beating rat cardiac myocytes before and after treatment with IPHC (isoproterenol hydrochloride). Full phase and topographic data were obtained at 15 frames/sec. Live cells were prepared and grown on either #1 coverslips or highly reflective slides. Cells cultures were placed into a Bioptechs FCS3 perfusion chamber and kept at 37°C during measurement. Videos of cell optical thickness topography were generated and processed to obtain relative optical volume. These results show dramatically increased rate and strength of contractions with application of IPHC.View Large Image | View Hi-Res Image | Download PowerPoint Slide