Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro
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...
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doaj-2e8276b9d9df494f895d7b50202740cc2021-03-22T12:39:26ZengElsevierNeurobiology of Disease1095-953X2013-06-0154158168Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitroVictor Tapias0J. Timothy Greenamyre1Simon C. Watkins2Department of Neurology, University of Pittsburgh, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, USA; Correspondence to: V. Tapias, University of Pittsburgh, 3501 Fifth Avenue, Suite 7045, Pittsburgh, PA 15260, USA. Fax: +1 412 648 9766.Department of Neurology, University of Pittsburgh, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, USA; Pittsburgh VA Healthcare System, University of Pittsburgh, USA; Correspondence to: J.T. Greenamyre, University of Pittsburgh, 3501 Fifth Avenue, Suite 7039, Pittsburgh, PA 15260, USA. Fax: +1 412 648 9766.Center for Biologic Imaging, University of Pittsburgh, USA; Department of Cell Biology and Physiology, University of Pittsburgh, USAQuantitation 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.http://www.sciencedirect.com/science/article/pii/S0969996112003816NeuroprotectionNeurodegenerationNeurotoxicityRotenoneNeuronNeurites |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Victor Tapias J. Timothy Greenamyre Simon C. Watkins |
spellingShingle |
Victor Tapias J. Timothy Greenamyre Simon C. Watkins Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro Neurobiology of Disease Neuroprotection Neurodegeneration Neurotoxicity Rotenone Neuron Neurites |
author_facet |
Victor Tapias J. Timothy Greenamyre Simon C. Watkins |
author_sort |
Victor Tapias |
title |
Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro |
title_short |
Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro |
title_full |
Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro |
title_fullStr |
Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro |
title_full_unstemmed |
Automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro |
title_sort |
automated imaging system for fast quantitation of neurons, cell morphology and neurite morphometry in vivo and in vitro |
publisher |
Elsevier |
series |
Neurobiology of Disease |
issn |
1095-953X |
publishDate |
2013-06-01 |
description |
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. |
topic |
Neuroprotection Neurodegeneration Neurotoxicity Rotenone Neuron Neurites |
url |
http://www.sciencedirect.com/science/article/pii/S0969996112003816 |
work_keys_str_mv |
AT victortapias automatedimagingsystemforfastquantitationofneuronscellmorphologyandneuritemorphometryinvivoandinvitro AT jtimothygreenamyre automatedimagingsystemforfastquantitationofneuronscellmorphologyandneuritemorphometryinvivoandinvitro AT simoncwatkins automatedimagingsystemforfastquantitationofneuronscellmorphologyandneuritemorphometryinvivoandinvitro |
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