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Article: Morphometric analyses of retinal sections
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TitleMorphometric analyses of retinal sections
 
AuthorsChan, TF1
Chiu, K1
Lok, CKM1
Ho, WL1
So, KF1 2
Chang, RCC1 2
 
KeywordsCell size
Issue 60
Morphometric analysis
Neuroscience
Retina
Stereo investigator
Thickness
 
Issue Date2012
 
PublisherJournal of Visualized Experiments. The Journal's web site is located at http://wwwjovecom
 
CitationJournal Of Visualized Experiments, 2012 n. 60, article no. e3377 [How to Cite?]
DOI: http://dx.doi.org/10.3791/3377
 
AbstractMorphometric analyses of retinal sections have been used in examining retinal diseases. For examples, neuronal cells were significantly lost in the retinal ganglion cell layer (RGCL) in rat models with N-methyl-D-aspartate (NMDA)-induced excitotoxicity, retinal ischemia-reperfusion injury and glaucoma. Reduction of INL and inner plexiform layer (IPL) thicknesses were reversed with citicoline treatment in rats' eyes subjected to kainic acid-mediated glutamate excitotoxicity. Alteration of RGC density and soma sizes were observed with different drug treatments in eyes with elevated intraocular pressure. Therefore, having objective methods of analyzing the retinal morphometries may be of great significance in evaluating retinal pathologies and the effectiveness of therapeutic strategies. The retinal structure is multi-layers and several different kinds of neurons exist in the retina. The morphometric parameters of retina such as cell number, cell size and thickness of different layers are more complex than the cell culture system. Early on, these parameters can be detected using other commercial imaging software. The values are normally of relative value, and changing to the precise value may need further accurate calculation. Also, the tracing of the cell size and morphology may not be accurate and sensitive enough for statistic analysis, especially in the chronic glaucoma model. The measurements used in this protocol provided a more precise and easy way. And the absolute length of the line and size of the cell can be reported directly and easy to be copied to other files. For example, we traced the margin of the inner and outer most nuclei in the INL and formed a line then using the software to draw a 90 degree angle to measure the thickness. While without the help of the software, the line maybe oblique and the changing of retinal thickness may not be repeatable among individual observers. In addition, the number and density of RGCs can also be quantified. This protocol successfully decreases the variability in quantitating features of the retina, increases the sensitivity in detecting minimal changes. This video will demonstrate three types of morphometric analyses of the retinal sections. They include measuring the INL thickness, quantifying the number of RGCs and measuring the sizes of RGCs in absolute value. These three analyses are carried out with Stereo Investigator (MBF Bioscience MicroBrightField, Inc.). The technique can offer a simple but scientific platform for morphometric analyses. © 2012 Journal of Visualized Experiments.
 
ISSN1940-087X
2013 SCImago Journal Rankings: 0.475
 
DOIhttp://dx.doi.org/10.3791/3377
 
PubMed Central IDPMC3399493
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorChan, TF
 
dc.contributor.authorChiu, K
 
dc.contributor.authorLok, CKM
 
dc.contributor.authorHo, WL
 
dc.contributor.authorSo, KF
 
dc.contributor.authorChang, RCC
 
dc.date.accessioned2012-05-23T05:42:31Z
 
dc.date.available2012-05-23T05:42:31Z
 
dc.date.issued2012
 
dc.description.abstractMorphometric analyses of retinal sections have been used in examining retinal diseases. For examples, neuronal cells were significantly lost in the retinal ganglion cell layer (RGCL) in rat models with N-methyl-D-aspartate (NMDA)-induced excitotoxicity, retinal ischemia-reperfusion injury and glaucoma. Reduction of INL and inner plexiform layer (IPL) thicknesses were reversed with citicoline treatment in rats' eyes subjected to kainic acid-mediated glutamate excitotoxicity. Alteration of RGC density and soma sizes were observed with different drug treatments in eyes with elevated intraocular pressure. Therefore, having objective methods of analyzing the retinal morphometries may be of great significance in evaluating retinal pathologies and the effectiveness of therapeutic strategies. The retinal structure is multi-layers and several different kinds of neurons exist in the retina. The morphometric parameters of retina such as cell number, cell size and thickness of different layers are more complex than the cell culture system. Early on, these parameters can be detected using other commercial imaging software. The values are normally of relative value, and changing to the precise value may need further accurate calculation. Also, the tracing of the cell size and morphology may not be accurate and sensitive enough for statistic analysis, especially in the chronic glaucoma model. The measurements used in this protocol provided a more precise and easy way. And the absolute length of the line and size of the cell can be reported directly and easy to be copied to other files. For example, we traced the margin of the inner and outer most nuclei in the INL and formed a line then using the software to draw a 90 degree angle to measure the thickness. While without the help of the software, the line maybe oblique and the changing of retinal thickness may not be repeatable among individual observers. In addition, the number and density of RGCs can also be quantified. This protocol successfully decreases the variability in quantitating features of the retina, increases the sensitivity in detecting minimal changes. This video will demonstrate three types of morphometric analyses of the retinal sections. They include measuring the INL thickness, quantifying the number of RGCs and measuring the sizes of RGCs in absolute value. These three analyses are carried out with Stereo Investigator (MBF Bioscience MicroBrightField, Inc.). The technique can offer a simple but scientific platform for morphometric analyses. © 2012 Journal of Visualized Experiments.
 
dc.description.naturelink_to_OA_fulltext
 
dc.identifier.citationJournal Of Visualized Experiments, 2012 n. 60, article no. e3377 [How to Cite?]
DOI: http://dx.doi.org/10.3791/3377
 
dc.identifier.doihttp://dx.doi.org/10.3791/3377
 
dc.identifier.epagepii 3377 doi: 10.3791/3377
 
dc.identifier.hkuros199686
 
dc.identifier.issn1940-087X
2013 SCImago Journal Rankings: 0.475
 
dc.identifier.issue60, article no. e3377
 
dc.identifier.pmcidPMC3399493
 
dc.identifier.pmid22370760
 
dc.identifier.scopuseid_2-s2.0-84857817746
 
dc.identifier.spagepii 3377 doi: 10.3791/3377
 
dc.identifier.urihttp://hdl.handle.net/10722/146847
 
dc.identifier.volume60
 
dc.languageeng
 
dc.publisherJournal of Visualized Experiments. The Journal's web site is located at http://wwwjovecom
 
dc.publisher.placeUnited States
 
dc.relation.ispartofJournal of Visualized Experiments
 
dc.relation.referencesReferences in Scopus
 
dc.subjectCell size
 
dc.subjectIssue 60
 
dc.subjectMorphometric analysis
 
dc.subjectNeuroscience
 
dc.subjectRetina
 
dc.subjectStereo investigator
 
dc.subjectThickness
 
dc.titleMorphometric analyses of retinal sections
 
dc.typeArticle
 
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Author Affiliations
  1. The University of Hong Kong Li Ka Shing Faculty of Medicine
  2. The University of Hong Kong