Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells

Cells such as astrocytes and radial glia with many densely ramified, fine processes pose particular challenges for the quantification of structural motility. Here we report the development of a method to calculate a motility index for individual cells with complex, dynamic morphologies. This motilit...

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Main Authors: Mari Sild, Robert P. Chatelain, Edward S. Ruthazer
Format: Article
Language:English
Published: Wiley 2013-01-01
Series:Neural Plasticity
Online Access:http://dx.doi.org/10.1155/2013/853727
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author Mari Sild
Robert P. Chatelain
Edward S. Ruthazer
author_facet Mari Sild
Robert P. Chatelain
Edward S. Ruthazer
author_sort Mari Sild
collection DOAJ
description Cells such as astrocytes and radial glia with many densely ramified, fine processes pose particular challenges for the quantification of structural motility. Here we report the development of a method to calculate a motility index for individual cells with complex, dynamic morphologies. This motility index relies on boxcar averaging of the difference images generated by subtraction of images collected at consecutive time points. An image preprocessing step involving 2D projection, edge detection, and dilation of the raw images is first applied in order to binarize the images. The boxcar averaging of difference images diminishes the impact of artifactual pixel fluctuations while accentuating the group-wise changes in pixel values which are more likely to represent real biological movement. Importantly, this provides a value that correlates with mean process elongation and retraction rates without requiring detailed reconstructions of very complex cells. We also demonstrate that additional increases in the sensitivity of the method can be obtained by denoising images using the temporal frequency power spectra, based on the fact that rapid intensity fluctuations over time are mainly due to imaging artifact. The MATLAB programs implementing these motility analysis methods, complete with user-friendly graphical interfaces, have been made publicly available for download.
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spelling doaj-art-671eec6eeeba4f34a664dcfde1e975be2025-02-03T01:12:31ZengWileyNeural Plasticity2090-59041687-54432013-01-01201310.1155/2013/853727853727Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex CellsMari Sild0Robert P. Chatelain1Edward S. Ruthazer2Montreal Neurological Institute, 3801 University Street, McGill University Montreal, QC, H3A 2B4, CanadaDepartment of Physics, McGill University, CanadaMontreal Neurological Institute, 3801 University Street, McGill University Montreal, QC, H3A 2B4, CanadaCells such as astrocytes and radial glia with many densely ramified, fine processes pose particular challenges for the quantification of structural motility. Here we report the development of a method to calculate a motility index for individual cells with complex, dynamic morphologies. This motility index relies on boxcar averaging of the difference images generated by subtraction of images collected at consecutive time points. An image preprocessing step involving 2D projection, edge detection, and dilation of the raw images is first applied in order to binarize the images. The boxcar averaging of difference images diminishes the impact of artifactual pixel fluctuations while accentuating the group-wise changes in pixel values which are more likely to represent real biological movement. Importantly, this provides a value that correlates with mean process elongation and retraction rates without requiring detailed reconstructions of very complex cells. We also demonstrate that additional increases in the sensitivity of the method can be obtained by denoising images using the temporal frequency power spectra, based on the fact that rapid intensity fluctuations over time are mainly due to imaging artifact. The MATLAB programs implementing these motility analysis methods, complete with user-friendly graphical interfaces, have been made publicly available for download.http://dx.doi.org/10.1155/2013/853727
spellingShingle Mari Sild
Robert P. Chatelain
Edward S. Ruthazer
Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells
Neural Plasticity
title Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells
title_full Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells
title_fullStr Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells
title_full_unstemmed Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells
title_short Improved Method for the Quantification of Motility in Glia and Other Morphologically Complex Cells
title_sort improved method for the quantification of motility in glia and other morphologically complex cells
url http://dx.doi.org/10.1155/2013/853727
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AT robertpchatelain improvedmethodforthequantificationofmotilityingliaandothermorphologicallycomplexcells
AT edwardsruthazer improvedmethodforthequantificationofmotilityingliaandothermorphologicallycomplexcells