Abstract

Multiphoton imaging has been utilized to analyze in vivo immune cell dynamics over the last 15 years. Particularly, it has deepened the understanding of how immune responses are organized by immune cell migration and interactions. In this review, we first describe the following technical advances in recent imaging studies that contributed to the new findings on the regulation of immune responses and inflammation. Improved multicolor imaging of immune cell behavior has revealed that their interactions are spatiotemporally coordinated to achieve efficient and long-term immunity. The use of photoactivatable and photoconvertible fluorescent proteins has increased duration and volume of cell tracking, even enabling the analysis of inter-organ migration of immune cells. In addition, visualization of immune cell activation using biosensors for intracellular calcium concentration and signaling molecule activities has started to give further mechanistic insights. Then, we also introduce recent imaging analyses of interactions between immune cells and non-immune cells including endothelial, fibroblastic, epithelial, and nerve cells. It is argued that future imaging studies that apply updated technical advances to analyze interactions between immune cells and non-immune cells will be important for thorough physiological understanding of the immune system.Electronic supplementary materialThe online version of this article (doi:10.1007/s00424-016-1882-x) contains supplementary material, which is available to authorized users.

Highlights

  • The immune system is comprised of diverse immune cell types

  • Live imaging analysis with the 3D resolution at the cellular level using multiphoton laser excitation fluorescent microscopy provides vital information to understand in vivo cellular interactions

  • Earlier than in the immunology field, multiphoton microscopy was applied for live tissue imaging in the field of neuroscience in 1990s [10]

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Summary

Introduction

The immune system is comprised of diverse immune cell types. Different types of immune responses against various pathogens and environmental antigens need to employ different combinations of immune cells that interactively work together. Real-time imaging of immune cell behavior in excised, re-aggregated, or in vivo tissues using multiphoton or single-photon laser microscopy was first reported in 2002 to 2003 [7, 45, 46, 67].

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