Hara Manami, Noiseux Nicolas, Bindokas Vytas P
Department of Medicine, The University of Chicago, Chicago, IL 60637, USA.
Biotechniques. 2005 Sep;39(3):373-6. doi: 10.2144/05393ST05.
We describe a method to visualize green fluorescent protein (GFP)-labeled cells in intact organs through combined confocal and reflected laser light imaging. This method allows us a three-dimensional (3-D) view of specific cell types in situ. Imaging of tissues from transgenic mice in which the endothelial cells are labeled with GFP under the control of endothelial-specific tyrosine receptor kinase 2 (TIE2) shows the spatial distribution of the GFP-labeled endothelial cells in intact organs. We have used this method to examine the tissue necrosis in the intact heart and kidney resulting from myocardial and renal infarction. In myocardial infarction produced by surgically occluding the left anterior descending coronary artery, the border of the infarct was highly cellular and showed a disrupted endothelial network and scar tissue appearing as a dense layer of reflection. The induced renal infarction produced by ligating the renal artery in the pedicle showed a clear infarct border in the affected kidney. The 3-D reconstruction of specific cell types in the context of the surrounding tissues should be useful for studying the overall organization and the relationship between different structures in the intact organ in normal and disease states.
我们描述了一种通过共聚焦和反射激光成像相结合的方法,来可视化完整器官中绿色荧光蛋白(GFP)标记的细胞。这种方法使我们能够在原位对特定细胞类型进行三维(3-D)观察。对内皮细胞在内皮特异性酪氨酸受体激酶2(TIE2)控制下用GFP标记的转基因小鼠组织进行成像,显示了完整器官中GFP标记的内皮细胞的空间分布。我们已使用这种方法来检查心肌梗死和肾梗死导致的完整心脏和肾脏中的组织坏死。在通过手术阻断左前降支冠状动脉产生的心肌梗死中,梗死边界细胞高度密集,显示出内皮网络破坏,瘢痕组织呈现为致密的反射层。在肾蒂处结扎肾动脉诱导产生的肾梗死在受影响的肾脏中显示出清晰的梗死边界。在周围组织背景下对特定细胞类型进行三维重建,对于研究正常和疾病状态下完整器官的整体组织结构以及不同结构之间的关系应该是有用的。