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Comparing Imaging Depth of Intravital Lung Imaging Using Perfluorocarbon-Based Liquid Ventilation With Tissue

Pascal Detampel^1,2, Wolf Heusermann³, Katarzyna M Wojcik⁴

¹Department of Cell Biology and Anatomy, University of Calgary, Calgary, Alberta, Canada.

Journal of Biophotonics

|September 3, 2025

Summary

Partial liquid ventilation with perfluorocarbon (PFC) did not improve lung imaging depth but enabled nanoparticle delivery for observing lung epithelium interactions. This advances studies on inhaled particles and drug delivery.

Keywords:

in vivo intravital lung imaging mice multi photon microscopy nanoparticle delivery perfluorocarbon refractive index tissue optical clearing

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Area of Science:

Pulmonary physiology
Biomedical optics
Nanotechnology

Background:

Intravital lung imaging is crucial for studying nanoparticle deposition in the alveoli, relevant to air pollution and drug delivery.
Optical imaging depth is limited by the refractive index (RI) mismatch at the air-tissue interface.

Purpose of the Study:

To investigate strategies for improving optical imaging depth in the lung.
To evaluate the efficacy of partial liquid ventilation and optical tissue clearing for enhanced lung imaging and nanoparticle delivery.

Main Methods:

Evaluated partial liquid ventilation with oxygenated perfluorocarbon (PFC) to eliminate RI mismatch.
Assessed ex vivo optical tissue clearing with RI matching and lipid removal.
Observed nanoparticle delivery and interactions in vivo.

Main Results:

Partial liquid ventilation with PFC did not enhance imaging depth.
Ex vivo tissue clearing improved imaging depth only after removal of scattering lipids like pulmonary surfactant.
In vivo PFC ventilation enabled homogeneous nanoparticle delivery and real-time observation of lung epithelium interactions.

Conclusions:

Eliminating RI mismatch alone does not significantly improve in vivo lung imaging depth.
Partial liquid ventilation with PFC facilitates nanoparticle delivery for studying lung physiology and optimizing inhalation therapies.
This technique offers new possibilities for real-time observation of inhaled particle dynamics.