In heart muscle cells both longitudinal and transversal architectures along the main cell axises represent important building principles that overcome challenging local conditions considering cell size versus cellular metabolic, proteomic and energetic needs. Furthermore, the specific cell volume is occupied by myofilaments that are aligned with mitochondria and the endo/sarcoplasmic reticulum network (SR), which is tightly associated with a highly branched internal sarcolemmal membrane network, the T-tubules (TT). TTs distribute electrochemical signals and nutrients to intracellular Ca2+ release nanodomains and organelles that involve the junctional SR (jSR). Here we present fundamental properties of the highly branched T-tubule (TT) network as revealed by STED microscopy in isolated mouse cardiomyocytes. Healthy myocytes exhibit longitudinal and transversal components at similar frequencies. Yet, after myocardial infarction the specific micro-scale organization of TT membranes changes leading to a significant increase of longitudinal components and increased network branching, and overall to increased network complexity and spatial dysfunction. In addition, it remains unclear how TTs interface with the microtubule (MT) system and whether MTs contribute to the TT membrane remodeling described above. MTs have been functionally associated with the organization of SR terminal cisternae and sub-membrane scaffolds (in skeletal muscle) and CaV1.2 targeting to T-tubules (in heart). Therefore, we further examined the MT network architecture and pathological changes. We found predominantly longitudinal MT components with more complex network patterns in the periphery of cells (cortex) as compared to deep internal MTs. Our data suggest, that MT branching occurs at major functional points where dispersed Golgi organelles can be locally identified. Moreover we compare microtubule (MT) networks before and after myocardial remodeling after gradient-controlled aortic banding, which will be presented. In conclusion, quantitative analysis of membrane and protein networks showed important properties of spatial organization and suggest local mechanisms of remodeling that may contribute to intracellular Ca2+ release heterogeneity.