Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro

NEUROBIOLOGY OF DISEASE

卷:54

Article

Tapias, Victor^1,2  Greenamyre, J. Timothy^1,2,3  Watkins, Simon C.^4,5 

[1] Univ Pittsburgh, Dept Neurol, Pittsburgh, PA 15260 USA

[2] Univ Pittsburgh, Pittsburgh Inst Neurodegenerat Dis, Pittsburgh, PA 15260 USA

[3] Univ Pittsburgh, Pittsburgh VA Healthcare Syst, Pittsburgh, PA 15260 USA

[4] Univ Pittsburgh, Ctr Biol Imaging, Pittsburgh, PA 15260 USA

[5] Univ Pittsburgh, Dept Cell Biol & Physiol, Pittsburgh, PA 15260 USA

关键词: Neuroprotection;    Neurodegeneration;    Neurotoxicity;    Rotenone;    Neuron;    Neurites;    Morphology;    Quantification;   

DOI  :  10.1016/j.nbd.2012.11.018

来源: Elsevier

【 摘 要 】

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. (C) 2012 Elsevier Inc. All rights reserved.

【 授权许可】

Free