Abstract 612: Automated Analysis of Displacement From Intravital Multiphoton Microscopy in Mouse Ventricle

Abstract

Background: Multiphoton microscopy (MPM) has enabled in vivo time-lapse imaging of the heart that shows motion of cells within the tissue with micrometer resolution. We developed automated analysis techniques to quantify cellular motion from in vivo cardiac MPM images throughout the cardiac cycle. Methods: Intravital cardiac MPM of the beating mouse heart was performed on 26 week-old, C57Bl6 mice (n=6). Image volumes (100 μm deep) were acquired at 30 frames per second while recording the electrocardiogram and respiratory pressure. An image volume was reconstructed by assembling lines acquired nearest to a specified point in the cardio-respiratory phase space (Fig. a). Motion was calculated as the three-dimensional transformation required to register the reconstructed images to the image at the most stable cardiac phase. Results: Volumes were reconstructed in 50 intervals across the cardiac phase that show vasculature (intravenous Texas-red dextran, red) and cardiomyocytes (rhodamine 6G, cyan) moving across the field of view (Fig. b). Automated analysis indicated a maximum displacement occurring at 16 % (anterior-posterior), 36 % (base-apex) and 38 % (epi-endocardial) of the cardiac cycle defined by R-wave. Comparison by manual tracking of features across the cardiac cycle at a subset of phases (10 cardiac phases at peak exhalation) validated the automated measurement (Fig. c). Automated motion tracking shows superior performance in the spatial resolution and speed of analysis. Conclusions: We have shown a novel, fast, and accurate technique for characterizing cardiac motion from in vivo cardiac MPM to study the performance of the contractile cells in health and disease.