Imaging the beating heart in the mouse using intravital microscopy techniques

Real-time microscopic imaging of moving organs at single-cell resolution represents a major challenge in studying complex biology in living systems. Motion of the tissue from the cardiac and respiratory cycles severely limits intravital microscopy by compromising ultimate spatial and temporal imaging resolution. However, significant recent advances have enabled single-cell resolution imaging to be achieved in vivo. In this protocol, we describe experimental procedures for intravital microscopy based on a combination of thoracic surgery, tissue stabilizers and acquisition gating methods, which enable imaging at the single-cell level in the beating heart in the mouse. Setup of the model is typically completed in 1 h, which allows 2 h or more of continuous cardiac imaging. This protocol can be readily adapted for the imaging of other moving organs, and it will therefore broadly facilitate in vivo high-resolution microscopy studies.The project was funded by US National Institutes of Health (NIH) Contracts HHSN268201000044C and R01EB006432. This work was also supported by the National Research Foundation of Korea funded by the Korean Government (MSIP) 2012M3A6A3055694. A.D.A. was funded by the American Heart Association 14FTF20380185