Protocols using ImageJ for image quantification can be found online, but these protocols are generally directed towards the creation of specific plugins for ImageJ, are not from a validated publication or lack the necessary step-by-step protocol ( Many of these methods require either advanced coding/mathematical experience, use of expensive imaging software or specific hardware requirements that may not be feasible for every scientist ( There are several methods and publications ( Fedchenko and Reifenrath 2014 Jensen 2017) regarding using semi-quantitative IHC to compare or measure protein expression (
Imagej software nih free#
The semi-quantitative IHC involves using software such as free software ImageJ Fiji (version 1.2 WS Rasband, National Institute of Health, Bethesda, MD) to conduct deconvolution and downstream analysis. Semi-quantitative immunohistochemistry (IHC) is a powerful method for investigating protein expression and localization within tissues (Cregger 2006 Taylor and Levenson 2006 We anticipate this method will provide a practical guidance to the reader and make semi-quantification of proteins an easier task to include in publications. The protocol includes clear steps for a reader so that this method can be applied to many different proteins.
Imagej software nih how to#
The protocol uses semi-quantitative IHC of organic anion transporting polypeptide (OATP1B1) as an example, and gives detailed steps on how to deconvolute IHC images stained with hematoxylin and 3,3’-diaminobenzidine (DAB) and how to quantify their expression using ImageJ Fiji. The current protocol for the first time provides a detailed, step-by-step instruction of conducting semi-quantitative analysis of IHC images using ImageJ Fiji software so that researchers would be able to follow this single protocol to conduct their research.
Publications that use semi-quantification methods to quantify protein expression often don’t provide enough details in their methods section, which makes it difficult for the reader to reproduce their data. Currently, there is lack of an integrated protocol that includes a detailed procedure of how to measure or compare protein expression.
The semi-quantitative immunohistochemistry (IHC) involves using software such as free software ImageJ Fiji to conduct deconvolution and downstream analysis. Semi-quantitative IHC is a powerful method for investigating protein expression and localization within tissues.
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