Intravital Multiphoton Microscopy for Dissecting Cellular Dynamics in Arthritic Inflammation and Bone Destruction

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1142)

Abstract

Osteoclasts are giant bone-resorbing polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. They play critical roles not only in normal bone homeostasis (remodeling) but also in the pathogenesis of bone-destructive disorders such as osteoporosis and rheumatoid arthritis. However, how the activity of mature osteoclasts is regulated in vivo remains unclear. To answer this question, we recently developed an advanced imaging system to visualize living bone tissues with intravital multiphoton microscopy. Using this system, we succeeded in visualization of mature osteoclasts in living bones.

We herein describe the detailed methodology for visualizing bone resorption of mature osteoclasts in living bone marrow and joints using intravital multiphoton microscopy. This approach would be beneficial for studying the cellular dynamics in arthritic inflammation and bone destruction in vivo and would thus be useful for evaluating novel anti-bone-resorptive drugs.

Key words

Intravital imaging Multiphoton microscopy Osteoclast Bone resorption Th17 cell 
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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Immunology and Cell Biology, Graduate School of Medicine and Frontier BiosciencesOsaka UniversitySuitaJapan

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