Automated analysis of images for molecular quantification in immunohistochemistry

Heliyon
Ramon GuiradoJuan Nacher

Abstract

The quantification of the expression of different molecules is a key question in both basic and applied sciences. While protein quantification through molecular techniques leads to the loss of spatial information and resolution, immunohistochemistry is usually associated with time-consuming image analysis and human bias. In addition, the scarce automatic software analysis is often proprietary and expensive and relies on a fixed threshold binarization. Here we describe and share a set of macros ready for automated fluorescence analysis of large batches of fixed tissue samples using FIJI/ImageJ. The quantification of the molecules of interest are based on an automatic threshold analysis of immunofluorescence images to automatically identify the top brightest structures of each image. These macros measure several parameters commonly quantified in basic neuroscience research, such as neuropil density and fluorescence intensity of synaptic puncta, perisomatic innervation and col-localization of different molecules and analysis of the neurochemical phenotype of neuronal subpopulations. In addition, these same macro functions can be easily modified to improve similar analysis of fluorescent probes in human biopsies for diagnostic purp...Continue Reading

Citations

Dec 4, 2019·The Journal of Comparative Neurology·Hector CarcellerJuan Nacher
Dec 19, 2019·Frontiers in Physiology·Jenna M KastenschmidtS Armando Villalta
Feb 19, 2020·Annals of Laboratory Medicine·Yousun ChungSun Young Kong
Oct 30, 2020·Journal of Pain Research·Chong PengWen Shen
Jul 26, 2019·Neurobiology of Disease·Udaiyappan JanakiramanMark A Nelson
Apr 4, 2021·Journal of Clinical Medicine·Carlos Arias-CabralesJulio Pascual
Jun 3, 2021·International Journal of Molecular Sciences·Simona CovielloJuan Nacher
Jul 12, 2021·Neurobiology of Disease·Carmen Navarro-GonzalezPietro Fazzari
Oct 14, 2021·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Cheng WangKimberley S Samkoe

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