Juan Sebastian Yakisich  

Virtual blots for teaching image analysis with ImageJ

JSY - MolBio


      The quantification of the relative expression of macromolecules (e.g. protein  by western blot)  is a routine technique in research laboratories but could be challenging for students. ImageJ is a imaging processing and analysis software written in Java widely used for quantification purposes ( http://rsb.info.nih.gov/ij/features.html ). However, there are few tutorials describing methods to quantify bands produced by Western, Northern or Southern Blots.  

      In this tutorial we will use ImageJ as a tool to introduce students in the quantitative analysis of virtual bands resembling Western , Northern and Southern Blots that are routine techniques to study the expression of proteins, RNA and DNA respectively.

     Later on, we will apply these concepts to Dot Blots.

     Finally, these principles can be used to quantity bands and dots generated in research labs.



1) The ImageJ software (1.46r version)  can be downloaded from the NIH website. (http://imagej.nih.gov/ij/download.html ) once installed in your computer it will looks like is shown in Fig 1).

2) Basic knowledge of PowerPoint and Excel


ImageJ (Main page, Fig. 1)









Figure 1


This tutorial is divided in several sections that can be navigated through the main menu found at the top of each webpage.  (Fig. 2A-B)













Figure 2A


Figure 2b



Additional Resources:

1) http://lukemiller.org/index.php/2010/11/analyzing-gels-and-western-blots-with-image-j/ . A tutorial developed by MT Miller using ImageJ, virtual bands but a different quantification method.



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