Atomic Force Microscopy (AFM) has emerged these recent years as a multifunctional toolbox for studying biological samples in physiological conditions. Although its use has spread among biologists community, cardiology remains a scientific field where not been extensively used yet. Heart diseases are nowadays a major human threat, and cause the death of millions of people each year. A convergent point to all heart diseases seems to be related to the defect of the cardiomyocyte, the contractile unit of the he reason, many scientists got interested in this cell type. However, very few studies use a technology such as AFM and its derivatives (force spectroscopy, multiparametric imaging) to explore this cell. The aim of this review is thus to give a comprehensive an interest of the biophysical approach made possible by AFM studies. We will show how AFM has been and can be used to study fix living cardiomyocytes, and, how combined with other types of microscopy, it can help getting a better understanding o pathologies or drugs. This review is the first dedicated to the use of AFM technics in cardiology, and gives new insights in fundamental questions surrounding cardiomyocytes, that can be answered using such a technology.