ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis

Co-localization analysis is pivotal for understanding protein interactions in biomedical research, yet existing ImageJ and FIJI plug-ins often lack automated multi-channel capabilities, impeding throughput and introducing potential user bias. We introduce ICOBA (Iterative Channel Overlay Batch Analy...

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Main Authors: Tyler J. Rolland, Emily R. Hudson, Luke A. Graser, Brian R Weil
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:SoftwareX
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352711025000615
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author Tyler J. Rolland
Emily R. Hudson
Luke A. Graser
Brian R Weil
author_facet Tyler J. Rolland
Emily R. Hudson
Luke A. Graser
Brian R Weil
author_sort Tyler J. Rolland
collection DOAJ
description Co-localization analysis is pivotal for understanding protein interactions in biomedical research, yet existing ImageJ and FIJI plug-ins often lack automated multi-channel capabilities, impeding throughput and introducing potential user bias. We introduce ICOBA (Iterative Channel Overlay Batch Analysis), a freely available ImageJ macro designed to streamline and standardize co-localization workflows across large image datasets. As a demonstration of the workflow and to validate its performance, cardiac fibroblasts were immunostained and imaged on a Leica DMi8 microscope, with .tiff files exported for processing. Compared to traditional manual approaches, ICOBA demonstrated significantly faster single-channel and two-channel processing times without sacrificing quantitative accuracy. By leveraging ImageJ's built-in “record” functionality and a customizable macro script, ICOBA accommodates variable staining conditions and threshold parameters, ensuring both reproducibility and flexibility. These attributes make ICOBA a versatile solution for high-throughput, multi-channel co-localization analyses across diverse research fields, from routine lab applications to advanced tissue-imaging studies.
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spelling doaj-art-b12f7e27dc0e4882950f317fe5bc73cf2025-08-20T03:05:50ZengElsevierSoftwareX2352-71102025-05-013010209410.1016/j.softx.2025.102094ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysisTyler J. Rolland0Emily R. Hudson1Luke A. Graser2Brian R Weil3Department of Physiology & Biophysics at the University at Buffalo and the VA WNY Healthcare System, Buffalo, NY, USADepartment of Physiology & Biophysics at the University at Buffalo and the VA WNY Healthcare System, Buffalo, NY, USADepartment of Pharmacology & Toxicology at the University at Buffalo, Buffalo, NY, USADepartment of Physiology & Biophysics at the University at Buffalo and the VA WNY Healthcare System, Buffalo, NY, USA; Corresponding author at: Department of Physiology and Biophysics, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo, Clinical Translational Research Center, Suite 7030, 875 Ellicott Street, Buffalo, NY 14203.Co-localization analysis is pivotal for understanding protein interactions in biomedical research, yet existing ImageJ and FIJI plug-ins often lack automated multi-channel capabilities, impeding throughput and introducing potential user bias. We introduce ICOBA (Iterative Channel Overlay Batch Analysis), a freely available ImageJ macro designed to streamline and standardize co-localization workflows across large image datasets. As a demonstration of the workflow and to validate its performance, cardiac fibroblasts were immunostained and imaged on a Leica DMi8 microscope, with .tiff files exported for processing. Compared to traditional manual approaches, ICOBA demonstrated significantly faster single-channel and two-channel processing times without sacrificing quantitative accuracy. By leveraging ImageJ's built-in “record” functionality and a customizable macro script, ICOBA accommodates variable staining conditions and threshold parameters, ensuring both reproducibility and flexibility. These attributes make ICOBA a versatile solution for high-throughput, multi-channel co-localization analyses across diverse research fields, from routine lab applications to advanced tissue-imaging studies.http://www.sciencedirect.com/science/article/pii/S2352711025000615ICOBAImageJ macroCo-localizationImage analysis
spellingShingle Tyler J. Rolland
Emily R. Hudson
Luke A. Graser
Brian R Weil
ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis
SoftwareX
ICOBA
ImageJ macro
Co-localization
Image analysis
title ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis
title_full ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis
title_fullStr ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis
title_full_unstemmed ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis
title_short ICOBA: A highly customizable iterative imagej macro for optimization of image co-localization batch analysis
title_sort icoba a highly customizable iterative imagej macro for optimization of image co localization batch analysis
topic ICOBA
ImageJ macro
Co-localization
Image analysis
url http://www.sciencedirect.com/science/article/pii/S2352711025000615
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AT lukeagraser icobaahighlycustomizableiterativeimagejmacroforoptimizationofimagecolocalizationbatchanalysis
AT brianrweil icobaahighlycustomizableiterativeimagejmacroforoptimizationofimagecolocalizationbatchanalysis