Intravital microscopy of the mouse brain microcirculation using a closed cranial window.
Intravital microscopy of the mouse brain microcirculation using a closed cranial window.
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DOI：
10.3791/2184
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发表时间：
2010-11-18
期刊：
影响因子：
--
通讯作者：
Carvalho, Leonardo J M
中科院分区：
文献类型：
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作者：
Cabrales, Pedro;Carvalho, Leonardo J M
该实验模型的目的是评估小鼠软脑膜微循环在急性和慢性，生理和病理生理血流动力学，炎症和代谢条件下，使用体内荧光显微镜。在小鼠的左顶叶-枕叶皮质上放置闭合的颅窗。局部微循环通过窗口使用epi和荧光照明进行真实的时间记录，并且进行血管直径和红细胞（RBC）速度的测量。使用实时互相关和/或荧光标记的红细胞测量RBC速度。白细胞和血小板粘附到软脑膜血管和评估灌注和血管渗漏的荧光标记物，如白蛋白-FITC和抗CD 45-TxR抗体的帮助下进行。微循环可以在几天内重复录像。我们第一次使用了闭窗脑活体显微镜来研究软脑膜微循环在伯氏疟原虫ANKA感染小鼠过程中的动态变化，并表明CM的表达与微循环功能障碍有关，其特征在于血管收缩，血流量显著减少，最终血管塌陷。
This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, inflammatory and metabolic conditions, using in vivo fluorescence microscopy. A closed cranial window is placed over the left parieto-occipital cortex of the mice. Local microcirculation is recorded in real time through the window using epi and fluorescence illumination, and measurements of vessels diameters and red blood cell (RBC) velocities are performed. RBC velocity is measured using real-time cross-correlation and/or fluorescent-labeled erythrocytes. Leukocyte and platelet adherence to pial vessels and assessment of perfusion and vascular leakage are made with the help of fluorescence-labeled markers such as Albumin-FITC and anti-CD45-TxR antibodies. Microcirculation can be repeatedly video-recorded over several days. We used for the first time the close window brain intravital microscopy to study the pial microcirculation to follow dynamic changes during the course of Plasmodium berghei ANKA infection in mice and show that expression of CM is associated with microcirculatory dysfunctions characterized by vasoconstriction, profound decrease in blood flow and eventually vascular collapse.