Visualizing inflammation at high spatio-temporal resolution: A novel minimally invasive approach to intravital imaging with an Airy Beam Light Sheet microscope (Aurora)

Session

Poster Session

Authors

Rebekka I Stegmeyer (2), Malte Stasch (2), Daniel Olesker (1, 3), Tom Mitchell (1), Jonathan Taylor (3), Dietmar Vestweber (2), Stefan Volkery (2), Neveen A. Hosny (1)

Affiliations

1. M Squared Life Limited
2. Max-Planck-Institute for Molecular Biomedicine
3. University of Glasgow

Keywords

intra vital microscopy, light sheet microscopy, improved temporal resolution

Abstract text

Intravital microscopy has emerged as a powerful imaging tool and revolutionized our ability to observe and comprehend rapid physiological phenomena within inflamed tissue environments in vivo. These include processes such as vascular permeability and leukocyte migration. In particular, the interactions between leukocytes and the vascular endothelium can be effectively characterized within the natural environment of the murine cremaster muscle.

Here, we introduce a novel microscopy approach with an Airy Beam Light Sheet 

Microscope, offering significant advantages over conventional confocal microscopy systems previously employed. Notably, this innovative technique offers isotropic resolution and enhanced recording speed, all while encompassing a larger field of view. Furthermore, through the implementation of a less invasive surgical procedure, we have successfully reduced side effects such as bleeding, muscle twitching, and surgical inflammation. However, the enhanced acquisition speed requires tissue stability to prevent imaging distortions. To address this challenge, we have developed a motion artifact correction algorithm to compensate for tissue movement induced by respiration, particularly relevant in the dynamic environment of the murine cremaster muscle. By employing this innovative technique enhancing the stability of intravital time-series, we have achieved more comprehensive 3D time-lapse images of the cremaster microcirculation. This enables detailed observation of the migration of leukocytes into the surrounding tissue with significantly improved temporal resolution and provides valuable insights for biomedical research.