In vivo multiphoton microscopy of deep brain tissue
In vivo multiphoton microscopy of deep brain tissue
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DOI：
10.1152/jn.01007.2003
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发表时间：
2004-04-01
影响因子：
2.5
通讯作者：
Webb, WW
中科院分区：
文献类型：
--
作者：
Levene, MJ;Dombeck, DA;Webb, WW
尽管荧光显微镜已被证明是生物学中最强大的工具之一，但它对完整动物的应用仅限于对表面以下数百微米的成像。动物的其余部分在没有切除组织或进行广泛手术的情况下逃避了显微镜水平的研究。然而，在完整动物中以亚细胞分辨率成像的能力使得能够实现对于理解适当功能可能至关重要的背景设置。诸如疾病诊断和光学活检的临床应用可以受益于微创体内方法。具有针状尺寸的梯度折射率（GRIN）透镜可以将高质量图像传输到距离物平面许多厘米的地方。在这里，我们表明，通过GRIN透镜的多光子显微镜，使微创，亚细胞分辨率几毫米的麻醉，完整的动物，我们目前在体内的皮质层V和海马在麻醉的Thy 1-YFP行H小鼠的图像。显微血管造影从深毛细血管和血管含有荧光素葡聚糖和量子点标记的血清在野生型小鼠脑也被证明。
Although fluorescence microscopy has proven to be one of the most powerful tools in biology, its application to the intact animal has been limited to imaging several hundred micrometers below the surface. The rest of the animal has eluded investigation at the microscopic level without excising tissue or performing extensive surgery. However, the ability to image with subcellular resolution in the intact animal enables a contextual setting that may be critical for understanding proper function. Clinical applications such as disease diagnosis and optical biopsy may benefit from minimally invasive in vivo approaches. Gradient index (GRIN) lenses with needle-like dimensions can transfer high-quality images many centimeters from the object plane. Here, we show that multiphoton microscopy through GRIN lenses enables minimally invasive, subcellular resolution several millimeters in the anesthetized, intact animal, and we present in vivo images of cortical layer V and hippocampus in the anesthetized Thy1-YFP line H mouse. Microangiographies from deep capillaries and blood vessels containing fluorescein-dextran and quantum dot-labeled serum in wild-type mouse brain are also demonstrated.