Super-resolution fluorescence imaging to study cardiac biophysics: α-actinin distribution and Z-disk topologies in optically thick cardiac tissue slices.

摘要: A major motivation for the use of super-resolution imaging methods in investigation cardiac biophysics has been insight from biophysical considerations and detailed mathematical modeling that spatial structure protein organisation at scale nanometres can have enormous implications calcium signalling muscle. We illustrate dSTORM based optically thick (∼10 μm) tissue slices rat ventricular to visualize proteins Z-disk compare those images with confocal (diffraction-limited) as well electron microscopy (EM) data which still provides a benchmark terms resolution. α-actinin is an abundant target effectively defines striated muscle reference other Z-line transverse tubules. Using labelling very outlines contractile machinery we used study properties Z-disks distribution itself. determined local diameters myo-fibrillar non-myofibrillar space using labelling. Comparison between myofibrillar masks suggested was able segment myofibrils accurately while approaches were not always distinguish neighbouring bundles resulted overestimated diameters. The increased resolution qualitatively new information improve our understanding biophysics. Nevertheless, conventional diffraction-limited important role play correlative data.