基于双光子显微镜技术的脑微血栓动物模型的建立与评价

Induction and evaluation of microthrombosis stroke in mice using two-photon microscopy

利用正置双光子显微镜系统和荧光探针标记技术,构建脑深部微血管血栓动物模型。麻醉小鼠,制作活体小鼠颅骨开窗样本,尾静脉注射血浆标记物fluorescein isothiocyanate-dextran(FITC-dextran),通过z序列扫描检测脑内血管系统的三维分布;通过飞秒激光定点照射脑内目标血管建立微血栓动物模型;利用线扫描测量目标血管形成血栓后的血流速度,评价微血栓形成的效果。通过双光子显微镜可以探测到脑内500μm处的血管分布和走向,图像清晰且信噪比高;通过光刺激能够在脑内形成稳定的微血栓;微血栓形成后血流速度明显变慢。利用双光子显微镜诱导微血栓技术初步建立,为基础研究和医药应用提供了微血栓动物模型。

An animal model of deep microvascular thrombosis was constructed using an upright two-photon microscope system and a fluorescent probe labeling technique.The mice were anesthetized and skull window sample was made.The plasma marker fluorescein isothiocyanate-dextran(FITC-dextran)was injected through the tail vein.The three-dimensional distribution of the vasculature in the brain was detected by z-sequence scanning;the brain was irradiated by femtosecond laser light.Deep target vessels(100μm±10μm in the brain)were established to establish an animal model of microthrombosis;the blood flow velocity after thrombosis was measured by a line scan to evaluate the effect of microthrombosis.The two-photon microscope can detect the distribution and direction of blood vessels in the brain at 500μm.The image is clear and the signal-to-noise ratio is high.A stable microthrombus can be formed in the brain through light stimulation;the blood flow velocity is significantly slowed after microthrombosis.Micro-thrombosis induced by two-photon microscopy was initially established to provide micro-thrombotic animal models for basic research and medical applications.