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.
Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Claudio
C
Vinegoni
Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.|Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Aaron D
AD
Aguirre
Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.|School of Electrical Engineering, Hanyang University, Ansan, Republic of Korea.
Sungon
S
Lee
Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Ralph
R
Weissleder
D000818:Animals; D003199:Computer Systems; D006321:Heart / Q000502:physiology*; D000069416:Intravital Microscopy / Q000379:methods*; D051379:Mice; D009206:Myocardium / Q000166:cytology*; D061848:Optical Imaging / Q000379:methods*