For informations about installation of the image analysis tool, access the “Simulator” tab
Step 1: Open ImageJ by double clicking ImageJ icon from the desktop or one may go to start programme ImageJ on the windows operating system.
The image will open in a new window as shown below
Step 2: To analyze an Image into ImageJ, we have to import the image into ImageJ. To perform that, Go to files ---> Open. A popup window will appear. Use Choose button to choose your respective images into ImageJ software. The imported picture will shown in a new window
Step 3: Convert the images into "grayscale” (go to image ---> Type ---> RGB stack).
Step 4: Adjust the RGB grey scale manually by moving the lower slider
Step 5: Use the Image ---> Stacks ---> Make Montage command to view all three channels at the same time.
Step 6: Adjust the Image ---> Adjust ---> Threshold tool. The threshold must be manually adjusted by moving the lower slider until the stained liver tissue is highlighted in red.
Step 7: Set threshold level and press “OK”
Step 8: Choose AnalyzeSet Measurements. The Set Measurements window opens. Select Area, Area fraction, Limit to Threshold and Display label measurements. Then click OK.
Step 9: To measure the selected area. Press Analyze ---> Measure. The Area and mean area of the stained samples appears in a new window. Repeat the above steps until all bands have been measured. Then copy all the values to a spreadsheet. The quantified images are now ready for data analysis in the spreadsheet program.
“Results” window indicates the Area and Percentage area of the stained cells. Here 23.2555% of the area contains the stained cells.