Intubation-free in vivo imaging of the tracheal mucosa using two-photon microscopy

Tibor Z Veres,Sirpa Jalkanen,Marko Tirri,Tamás Kopcsányi,Armin Braun,Marko Salmi,Ronald N Germain,Masayuki Miyasaka

doi:10.1038/s41598-017-00769-6

Abstract

The mucosal layer of conducting airways is the primary tissue exposed to inhaled microorganisms, allergens and pollutants. We developed an in vivo two-photon microscopic approach that allows performing dynamic imaging studies in the mouse trachea, which is a commonly used in vivo model of human small-diameter bronchi. By providing stabilized access to the tracheal mucosa without intubation, our setup uniquely allows dynamic in vivo imaging of mucociliary clearance and steady-state immune cell behavior within the complex airway mucosal tissue.

Highlights

During the last decade, multi-photon intravital microscopy (MP-IVM) has become the “gold standard” technique for imaging biological processes in vivo and in real time at single-cell resolution[1]
We were aided by micro-CT to better understand the in situ anatomy of the surgically exposed trachea (Supplementary Figure 2). 3D-modeling in combination with Computer Numeric Control (CNC) precision milling or 3D-printing (Supplementary Figure 3) were used for the rapid production of prototypes including very small parts with well-defined specific shapes and dimensions
The mucosa of the conducting airways represents an important anatomical/physiological barrier to the outside environment. This barrier function is maintained by the airway surface liquid (ASL) containing mucus; the ciliated, pseudostratified epithelium; and by the strategically localized intraepithelial dendritic cells (IE-DCs) network

Summary

Introduction

Multi-photon intravital microscopy (MP-IVM) has become the “gold standard” technique for imaging biological processes in vivo and in real time at single-cell resolution[1]. The new approach helps to minimize inflammation and vascular leakage, preserves steady-state immune cell behavior and allows direct observation of physiological mucociliary clearance.

Results

Conclusion