Image Analysis with ImageJ NOTE: There is an tutorial video available to step you through ‘ImageJ’ Video tutorial image processing with ImageJ. available on D2L Quantifying changes in the of gene expression is a challenging task, even with the most pattern sophisticated microscope analysis software available. While it may be easy to qualitatively describe the change in location of a GFP signal, we will be quantifying this change in lab today through determining a ratio of nuclear to cytoplasmic GFP expression. To accomplish this task, we will use ImageJ, a free, public domain Java image-processing program that is commonly used by research scientists. The image processing protocol is provided below. Open ImageJ through the Departmental Applications folder at the bottom of the computer screen. The ImageJ toolbar should be visible, but you will have to open each of your images individually through the ImageJ File Menu bar (File Open…). Begin by opening BOTH the light microscopy and epifluorescence microscopy images from your initial experimental time point. Place the images side-by-side so that you can easily see both images.
Image Analysis with ImageJ NOTE: There is an ‘ImageJ’ tutorial video available to step you through image processing with ImageJ. Video tutorial available on D2L Quantifying changes in the pattern of gene expression is a challenging task, even with the most sophisticated microscope analysis software available. While it may be easy to qualitatively describe the change in location of a GFP signal, we will be quantifying this change in lab today through determining a ratio of nuclear to cytoplasmic GFP expression. To accomplish this task, we will use ImageJ, a free, public domain Java image-processing program that is commonly used by research scientists. The image processing protocol is provided below. Open ImageJ through the Departmental Applications folder at the bottom of the computer screen. The ImageJ toolbar should be visible, but you will have to open each of your images individually through the ImageJ File Menu bar (File Open…). Begin by opening BOTH the light microscopy and epifluorescence microscopy images from your initial experimental time point. Place the images side-by-side so that you can easily see both images.
imagej_instructions.docx