Sub-microscopic analysis of t-tubule geometry in living cardiac ventricular myocytes using a shape-based analysis method

Abstract Transverse-axial tubules (TTs) are key structures involved in cardiac excitation-contraction coupling and can become deranged in disease. Although optical measurement of TTs is frequently employed to assess TT abundance and regularity, TT dimensions are generally below the diffraction limit of optical microscopy so determination of tubule size is problematic. TT diameter was measured by labeling both local surface membrane area and volume with fluorescent probes (FM4-64 and calcein, respectively), correcting image asymmetry by image processing and using the relationship between surface area and volume for a geometric primitive. This method shows that TTs have a mean (± SEM) diameter of 356 ± 18 nm in rabbit and 169 ± 15 nm in mouse ( p p