使用ImageJ分析WesternBlot..doc

使用ImageJ分析Western Blot? 2011-11-29 20:13:54|??分类： Bioinformatics |??标签： |字号大中小?订阅 Analyzing gels and western blots with ImageJ 2011-07-29 18:02 ? 来源：丁香园 ? 点击次数： 496 关键词： ImageJ 数据分析 分享到： 收藏夹 腾讯微博 新浪微博 开心网 The following information is an updated version of a method for using ImageJ to analyze western blots from a now-deprecated older page. Don’t use the alternate methods discussed on the old page, as they are subject to way too much user bias. A pdf copy of this page is available. ImageJ (/ij/index.html) can be used to compare the density (aka intensity) of bands on an agar gel or western blot. This tutorial assumes that you have carried your gel or blot through the visualization step, so that you have a digital image of your gel in .tif, .jpg, .png or other image formats (.tif would be the preferred format to retain the maximum amount of information in the original image). If you are scanning x-ray film on a flatbed scanner, make sure you use a scanner with the ability to scan transparencies (i.e. film). See the references at the end of this tutorial for a discussion of the various ways that you can screw this step up. The method outlined here uses the Gel Analysis method outlined in the ImageJ documentation: Gel Analysis. You may prefer to use it instead of the methods I outline below. There should be very little difference between the results obtained from the various methods. This version of the tutorial was created using ImageJ 1.42q on a Windows 7 64-bit install. 1. Open the image file using FileOpen in ImageJ. 2. The gel analysis routine requires the image to be a gray-scale image. The simplest method to convert to grayscale is to go to ImageType8-bit. Your image should look like Figure 1. 3. Choose the Rectangular Selections tool from the ImageJ toolbar. Draw a rectangle around the first lane. ImageJ assumes that your lanes run vertically (so individual bands are horizontal), so your rectangle should be tall and narrow to enclose a single lane. If you dr