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Imaging the beating heart in the mouse using intravital microscopy techniques
Vinegoni, Claudio,Aguirre, Aaron D,Lee, Sungon,Weissleder, Ralph Nature Publishing Group 2015 NATURE PROTOCOLS -ELECTRONIC EDITION- Vol.10 No.11
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.
Advanced Motion Compensation Methods for Intravital Optical Microscopy
Vinegoni, Claudio,Sungon Lee,Feruglio, Paolo Fumene,Weissleder, Ralph IEEE 2014 IEEE journal of selected topics in quantum electro Vol.20 No.2
<P>Intravital microscopy has emerged in the recent decade as an indispensible imaging modality for the study of the microdynamics of biological processes in live animals. Technical advancements in imaging techniques and hardware components, combined with the development of novel targeted probes and new mice models, have enabled us to address long-standing questions in several biology areas such as oncology, cell biology, immunology, and neuroscience. As the instrument resolution has increased, physiological motion activities have become a major obstacle that prevents imaging live animals at resolutions analogue to the ones obtained in vitro. Motion compensation techniques aim at reducing this gap and can effectively increase the in vivo resolution. This paper provides a technical review of some of the latest developments in motion compensation methods, providing organ specific solutions.</P>