An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.

Microglia are specialized immune cells of the brain. Upon insult, microglia initiate a cascade of cellular responses including a characteristic change in cell morphology. To study the dynamics of microglia immune response in situ, we developed an automated image analysis method that enables the quan...

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Main Authors: Cleopatra Kozlowski, Robby M Weimer
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3294422?pdf=render
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author Cleopatra Kozlowski
Robby M Weimer
author_facet Cleopatra Kozlowski
Robby M Weimer
author_sort Cleopatra Kozlowski
collection DOAJ
description Microglia are specialized immune cells of the brain. Upon insult, microglia initiate a cascade of cellular responses including a characteristic change in cell morphology. To study the dynamics of microglia immune response in situ, we developed an automated image analysis method that enables the quantitative assessment of microglia activation state within tissue based solely on cell morphology. Per cell morphometric analysis of fluorescently labeled microglia is achieved through local iterative threshold segmentation, which reduces errors caused by signal-to-noise variation across large volumes. We demonstrate, utilizing systemic application of lipopolysaccharide as a model of immune challenge, that several morphological parameters, including cell perimeter length, cell roundness and soma size, quantitatively distinguish resting versus activated populations of microglia within tissue comparable to traditional immunohistochemistry methods. Furthermore, we provide proof-of-concept data that monitoring soma size enables the longitudinal assessment of microglia activation in the mouse neocortex imaged via 2-photon in vivo microscopy. The ability to quantify microglia activation automatically by shape alone allows unbiased and rapid analysis of both fixed and in vivo central nervous system tissue.
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spelling doaj.art-d7673c7dd9bd4e41a48a36fc14e9a44b2022-12-22T00:00:23ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0172e3181410.1371/journal.pone.0031814An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.Cleopatra KozlowskiRobby M WeimerMicroglia are specialized immune cells of the brain. Upon insult, microglia initiate a cascade of cellular responses including a characteristic change in cell morphology. To study the dynamics of microglia immune response in situ, we developed an automated image analysis method that enables the quantitative assessment of microglia activation state within tissue based solely on cell morphology. Per cell morphometric analysis of fluorescently labeled microglia is achieved through local iterative threshold segmentation, which reduces errors caused by signal-to-noise variation across large volumes. We demonstrate, utilizing systemic application of lipopolysaccharide as a model of immune challenge, that several morphological parameters, including cell perimeter length, cell roundness and soma size, quantitatively distinguish resting versus activated populations of microglia within tissue comparable to traditional immunohistochemistry methods. Furthermore, we provide proof-of-concept data that monitoring soma size enables the longitudinal assessment of microglia activation in the mouse neocortex imaged via 2-photon in vivo microscopy. The ability to quantify microglia activation automatically by shape alone allows unbiased and rapid analysis of both fixed and in vivo central nervous system tissue.http://europepmc.org/articles/PMC3294422?pdf=render
spellingShingle Cleopatra Kozlowski
Robby M Weimer
An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.
PLoS ONE
title An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.
title_full An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.
title_fullStr An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.
title_full_unstemmed An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.
title_short An automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo.
title_sort automated method to quantify microglia morphology and application to monitor activation state longitudinally in vivo
url http://europepmc.org/articles/PMC3294422?pdf=render
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