Intravital three-photon microscopy allows visualization over the entire depth of mouse lymph nodes.

ABSTRACT: Intravital confocal microscopy and two-photon microscopy are powerful tools to explore the dynamic behavior of immune cells in mouse lymph nodes (LNs), with penetration depth of ~100 and ~300 μm, respectively. Here, we used intravital three-photon microscopy to visualize the popliteal LN through its entire depth (600-900 μm). We determined the laser average power and pulse energy that caused measurable perturbation in lymphocyte migration. Long-wavelength three-photon imaging within permissible parameters was able to image the entire LN vasculature in vivo and measure CD8⁺ T cells and CD4⁺ T cell motility in the T cell zone over the entire depth of the LN. We observed that the motility of naive CD4⁺ T cells in the T cell zone during lipopolysaccharide-induc

SUBMITTER: Choe K 

PROVIDER: S-EPMC9210714 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature