![]() ![]() The proposed method represents an affordable, accurate and reproducible approximation that could be used in the presence of limited resources availability.īackground subtraction Densitometry ImageJ Quantification Western blot.Ĭopyright © 2018 Elsevier B.V. In this work, we describe the use of office scanner coupled with the ImageJ software together with a new image background subtraction method for accurate Western blot quantification. It utilizes densitometry measurement of ImageJ and subtracts density of image background. Furthermore, the use of semi-quantitative software as ImageJ (Java-based image-processing and analysis software) is clearly increasing in different scientific fields. ImageJ Analysis Tutorial Part 2 (Densitometry and Area) K R. Western Blot densitometry analysis - macro tool for ImageJ 1.x What is it good for This tool provides a quick and dirty way to measure images of not necessarily straight lines of Western Blot films, dot blots and other silimar bio-scientific images. Your gel will look something like this (see below). Once the gel image is open, you can zoom in with 'Ctrl+' and zoom out with 'Ctrl-'. You can drag the image you want to open onto the ImageJ window. The lanes were always positioned at the center of the gel lane. Open ImageJ using the shortcut on the desktop. The size of the lane selection tool was 16 pixels wide (Figure 2a), equivalent to 30 of the total width of the well as suggested by Gassmann et al. Due to the technical knowledge required for densitometry analysis together with the resources availability, standard office scanners are often used for the imaging acquisition of developed Western blot films. The Gel Analyzer tool of ImageJ was used to determine the profiles of each lane of the gels. Western blot quantification method constitutes a critical step in order to obtain accurate and reproducible results. In most cases, the detection of interactions leads to an image containing numerous lines of spots that will be analyzed by comparing tables of intensity values. It constitutes a multistep method that allows the detection and/or quantification of proteins from simple to complex protein mixtures. Protein array is used to analyze protein expressions by screening simultaneously several protein-molecule interactions such as protein-protein and protein-DNA interactions. Since its first description, Western blot has been widely used in molecular labs. ![]()
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