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Seeing the wood for the trees:towards improved quantification of glial cells in central nervous system tissue

Neural Regeneration Research 2018年第9期13卷 1520-1523页

作者： Sinéad Healy Jill McMahon Una FitzGerald Galway Neuroscience Centre School of Natural SciencesNational University of Ireland

The following mini-review attempts to guide researchers in the quantification of fluorescently-labelled proteins within cultured thick or chromogenically-stained proteins within thin sections of brain *** follows from our examination of the utility of Fiji Image J thresholding and binarization *** how we identified the maximum intensity projection as the best of six tested for two dimensional（2 D）-rendering of three-dimensional（3 D） images derived from a series of z-stacked micrographs,the review summarises our comparison of 16 global and 9 local algorithms for their ability to accurately quantify the expression of astrocytic glial fibrillary acidic protein（GFAP）,microglial ionized calcium binding adapter molecule 1（IBA1） and oligodendrocyte lineage Olig2 within fixed cultured rat hippocampal brain *** application of these algorithms to chromogenically-stained GFAP and IBA1 within thin tissue sections,is also ***’s bio voxxel plugin allowed categorisation of algorithms according to their sensitivity,specificity accuracy and relative *** Percentile algorithm was deemed best for quantifying levels of GFAP,the Li algorithm was best when quantifying IBA expression,while the Otsu algorithm was optimum for Olig2 staining,albeit with over-quantification of oligodendrocyte number when compared to a stereological ***,GFAP and IBA expression in 3,3′-diaminobenzidine（DAB）/haematoxylin-stained cerebellar tissue was best quantified with Default,Isodata and Moments *** workflow presented in Figure 1 could help to improve the quality of research outcomes that are based on the quantification of protein with brain tissue.

关键词： organotypic brain slice culture glial cell quantification thresholding algorithms Fiji Image J bio voxxel plug-in stereology

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