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Updated: May 18, 2026

Microvascular Embolism Mouse Model for In Vivo Two-photon Microscopy Using Fluorescent Polystyrene Microspheres
Published on: November 21, 2025
In vivo microvascular mosaics show air embolism reduction after perfluorocarbon emulsion treatment
Ivo P Torres Filho1, Luciana N Torres, Bruce D Spiess
1Department of Emergency Medicine, Virginia Commonwealth University Reanimation Engineering Shock Center, Virginia Commonwealth University, Richmond, VA 23298, USA. ivoptf@msn.com
Abstract:
Massive arteriolar gas embolism (AGE) has never been evaluated in vivo using intravital microscopy and previous perfluorocarbon (PFC) emulsions were only effective in AGE when administered before AGE. We implemented a new system for quantitative studies of massive AGE using brightfield microscopy and tested a treatment with a third-generation PFC emulsion after massive AGE. We studied bubble dynamics in cremaster muscles from anesthetized rats after AGE was induced by direct air injection into the femoral artery ipsilateral to the studied muscle. Using a motorized microscope stage and a color camera, in vivo microvascular mosaics were produced on-line from over 2000 digital images to evaluate multiple networks in order to investigate the distribution, lodging, breaking, reduction and moving of 105 air bubbles in microvessels. Thirty minutes after PFC treatment, there was a reduction of 80% in bubble volume while untreated and saline-treated rats showed significantly smaller decreases of 33% and 40%, respectively (p<0.05). Air bubbles also dissolved into a larger number of smaller bubbles after PFC treatment. The proposed methodology may prove useful for rapid in vivo data acquisition from large networks. Since large air bubbles broke-up, decreased in length and volume, and moved toward smaller microvessels, the study provides quantitative data to support a mechanism by which PFC may improve tissue blood flow following massive AGE. The findings suggest that this new generation of PFC emulsions administered after severe AGE may reach compromised microvascular networks and provide help to alleviate microvascular obstruction by increasing air bubble reabsorption.

