Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro.
Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro.
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DOI：
10.1016/j.nbd.2012.11.018
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发表时间：
2013-06
影响因子：
6.1
通讯作者：
Watkins SC
中科院分区：
文献类型：
--
作者：
Tapias V;Greenamyre JT;Watkins SC
神经元定量使用立体方法减少偏差和系统误差，但耗时和劳动密集。目前缺乏准确的体外神经元定量方法；传统的方法在可靠性和应用方面受到限制。体细胞和神经突的形态特征是神经元表型和功能的一个关键方面，但在这种评估中常用的测定方法受到几个方法学缺陷的困扰。在这里，我们描述了自动化技术来量化神经元（或任何细胞类型，如星形胶质细胞）的数量和形态及其过程，具有高速和准确。使用电动舞台系统对脑组织中的神经元进行定量分析，其结果在统计学上可与立体学产生的结果相媲美。该方法随后适用于体外神经元和神经突生长的定量。为了确定我们方法的实用性，鱼藤酮被用作神经毒物，导致神经元形态学改变和细胞死亡、星形细胞激活和神经突丢失。重要的是，我们的技术在不到手工立体分析5-10%的时间内计算了大约8倍的神经元。
Quantitation of neurons using stereologic approaches reduces bias and systematic error, but is time-consuming and labor-intensive. Accurate methods for quantifying neurons in vitro are lacking; conventional methodologies are limited in reliability and application. The morphological properties of the soma and neurites are a key aspect of neuronal phenotype and function, but the assays commonly used in such evaluations are beset with several methodological drawbacks. Herein we describe automated techniques to quantify the number and morphology of neurons (or any cell type, e.g., astrocytes) and their processes with high speed and accuracy. Neuronal quantification from brain tissue using a motorized stage system yielded results that were statistically comparable to those generated by stereology. The approach was then adapted for in vitro neuron and neurite outgrowth quantification. To determine the utility of our methods, rotenone was used as a neurotoxicant leading to morphological changes in neurons and cell death, astrocytic activation, and loss of neurites. Importantly, our technique counted about 8 times as many neurons in less than 5–10% of the time taken by manual stereological analysis.
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影响因子：
2
作者：
Gundersen, HJG;Jensen, EBV;Nielsen, J
通讯作者：
Nielsen, J
影响因子：
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作者：
Jiang, Q;Yan, Z;Feng, J
通讯作者：
Feng, J
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作者：
Gao, HM;Hong, JS;Liu, B
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作者：
Ma, SY;Rinne, JO;Rinne, UK
通讯作者：
Rinne, UK
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11
作者：
Chen, P-S;Peng, G-S;Hong, J-S
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Hong, J-S