Abstract
Invasive microsurgical techniques are currently used to induce collateral vessel formation by resecting or ligating a limited number of arteries feeding a vascular network. The objective of this study was to develop a non‐invasive technique for selectively blocking a specified number of small arterioles, venules and capillaries in a dense microvascular network such as the mouse dorsal skin fold chamber. We used a multiphoton laser to selectively ablate small vessel fragments through the glass window of the chamber in transgenic FVB/N mice, which express GFP in Tie‐2‐positive cells and Ds red in α‐SMA‐positive cells. Blood perfusion was visualized by i.v. injection of a fluorescence vascular tracer. We assessed diameter, length, and blood flow changes and tracked cellular dynamics during remodeling using time‐lapse fluorescence microscopy and Optical Frequency‐Domain Imaging (OFDI). The microsurgery did not interrupt blood flow to neighboring vessels or cause detectable tissue bleeding or damage. After an initial decrease, the diameter of upstream and downstream vessels returned to pre‐surgery levels within 24hours. In the following days, endothelial cells migrated to repair ablated regions of arterioles, eventually restoring flow. The new techniques can be further applied to study microvascular changes in normal and pathological (e.g. tumor) microvascular environments. Supported by NIH R01CA149285.
Published Version
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