Continuous high-resolution observations of cell membrane would greatly aid the elucidation of the relationship between structure and function and facilitate the study of physiological processing in cell biology. However, high-resolution studying living neuron membrane structures and its functions is still a challenge in current nanobiology. The new developed Hoping Probe Ion Conductance Microscopy (HPICM) is designed for non-contact continuous high-resolution topographic imaging of living cells under physiological conditions. In this review, we concisely introduced the basic operation principle of HPICM and its applications in high spatial resolution imaging of two living neuron cell models, N-type SK-N-SH cells and NGF-differentiated sympathetic neuron-like PC12 cells. Combining HPICM with patch-clamp technique, we further investigated the functional ion-channel of under-differentiated neuron-like PC12 cells and demonstrated that NGF treatment promoted the outgrowth of neurites and increased the activity of TTX-sensitive sodium channel. All these results demonstrate that HPICM combined with patch-clamp technique offers high-resolution topographic imaging of living neurons with non-contact — making HPICM an ideal high-resolution imaging technique not to interact/interfere with living neurons during image acquisition, and provides detailed information about the relationship between membrane structures and ion-channel functions of living neurons at the same time, which has the potential to become a powerful microscopy for in-depth researching in neural nanobiology.