Combining confocal laser scanning microscopy with serial section reconstruction in the study of adult neurogenesis.
Combining confocal laser scanning microscopy with serial section reconstruction in the study of adult neurogenesis.
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DOI：
10.3389/fnins.2011.00070
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发表时间：
2011
影响因子：
4.3
通讯作者：
Peretto P
中科院分区：
文献类型：
--
作者：
Luzzati F;Fasolo A;Peretto P
目前成像技术的进步已经扩大了在大量固定和活标本中可视化小结构而无需切片的可能性。这些技术为研究成人脑神经元的可塑性提供了有价值的信息。然而，技术上的限制仍然阻碍了使用这些方法来研究位于远离脑室表面的神经源性区域，如实质神经源性壁龛，或由脑病变引起的分散的神经母细胞。在这里，我们提出了一种方法，结合联合收割机共聚焦激光扫描显微镜（CLSM）和连续切片重建，以重建大量的脑组织细胞的分辨率。在这种方法中，一系列的厚切片成像与CLSM和所得到的图像堆栈注册和3D重建。这种方法是基于现有的免费软件，可以在普通的实验室个人电脑上进行。通过使用这种技术，我们已经调查的形态和空间组织的一组doublecortin（DCX）+神经母细胞位于出生后的豚鼠的外侧纹状体。3D研究揭示了一个复杂的网络，长而分支不良的细胞突起，往往是成簇的，主要是沿着沿着内囊纤维束定向。这些数据支持CLSM连续切片重建作为一个可靠的替代方法来分析成虫萌发壁龛的细胞结构特征。
Current advances in imaging techniques have extended the possibility of visualizing small structures within large volumes of both fixed and live specimens without sectioning. These techniques have contributed valuable information to study neuronal plasticity in the adult brain. However, technical limits still hamper the use of these approaches to investigate neurogenic regions located far from the ventricular surface such as parenchymal neurogenic niches, or the scattered neuroblasts induced by brain lesions. Here, we present a method to combine confocal laser scanning microscopy (CLSM) and serial section reconstruction in order to reconstruct large volumes of brain tissue at cellular resolution. In this method a series of thick sections are imaged with CLSM and the resulting stacks of images are registered and 3D reconstructed. This approach is based on existing freeware software and can be performed on ordinary laboratory personal computers. By using this technique we have investigated the morphology and spatial organization of a group of doublecortin (DCX)+ neuroblasts located in the lateral striatum of the late post-natal guinea pig. The 3D study unraveled a complex network of long and poorly ramified cell processes, often fascicled and mostly oriented along the internal capsule fiber bundles. These data support CLSM serial section reconstruction as a reliable alternative to the whole mount approaches to analyze cyto-architectural features of adult germinative niches.
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影响因子：
46.9
作者：
通讯作者：
--
影响因子：
6.1
作者：
Kernie SG;Parent JM
通讯作者：
Parent JM
平铺 3D 显微图像采集的全局最佳拼接。
DOI：
10.1093/bioinformatics/btp184
发表时间：
2009-06-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Preibisch S;Saalfeld S;Tomancak P
通讯作者：
Tomancak P
影响因子：
13.9
作者：
Kriegstein A;Alvarez-Buylla A
通讯作者：
Alvarez-Buylla A
影响因子：
4.8
作者：
Rodriguez, A;Ehlenberger, D;Wearne, SL
通讯作者：
Wearne, SL