Abstract

Intravital imaging microscopy (i.e., imaging in live animals at microscopic resolution) has become an indispensable tool for studying the cellular micro-dynamics in cancer, immunology and neurobiology. High spatial and temporal resolution, combined with large penetration depth and multi-reporter visualization capability make fluorescence intravital microscopy compelling for heart imaging. However, tissue motion caused by cardiac contraction and respiration critically limits its use. As a result, in vitro cell preparations or non-contracting explanted heart models are more commonly employed. Unfortunately, these approaches fall short of understanding the more complex host physiology that may be dynamic and occur over longer periods of time. In this review, we report on novel technologies, which have been recently developed by our group and others, aimed at overcoming motion-induced artifacts and capable of providing in vivo subcellular resolution imaging in the beating mouse heart. The methods are based on mechanical stabilization, image processing algorithms, gated/triggered acquisition schemes or a combination of both. We expect that in the immediate future all these methodologies will have considerable applications in expanding our understanding of the cardiac biology, elucidating cardiomyocyte function and interactions within the organism in vivo, and ultimately improving the treatment of cardiac diseases.

Highlights

  • Despite recent improvements in health care, cardiovascular diseases are still responsible for a stunning 30% of deaths worldwide (Go et al, 2013)

  • In particular we introduce the concept of gated “sequential segmented microscopy” (SSM), passive and active stabilization schemes, and image-processing algorithms for automatic motion-artifacts removal

  • If we obtain a set of CC tables for a generic image sequence it is possible to determine all motionless segments, to combine them according to their time coordinates and to reconstruct motion artifact-free image

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Summary

Introduction

Despite recent improvements in health care, cardiovascular diseases are still responsible for a stunning 30% of deaths worldwide (Go et al, 2013). Intravital confocal and multiphoton microscopy imaging has provided profound insights into in vivo cell biology (Pittet and Weissleder, 2011; Ritsma et al, 2012) offering high spatial and temporal resolution as well as deep-penetration depth and multi-reporter visualization. These capabilities have in turn enabled the acquisition of cellular information under natural physiological conditions and offered unique opportunities to explore and investigate biology in living systems. In particular we introduce the concept of gated “sequential segmented microscopy” (SSM), passive and active stabilization schemes, and image-processing algorithms for automatic motion-artifacts removal

Stabilization Methodologies
Applications in Cardiac Imaging
Future Perspectives
Full Text
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