Software:Fiji ImageJ
Flow of preprocessing of image
1. Making of sequential image
2. Cutting out of analytical area
3. Making of background image
4. Making of difference picture
5. Making of gray scale image
6. Adjusting contrasts
1. Making of sequential image
ImageJ operation : file→import→image sequence
2. Cutting out of analytical area
ImageJ operation : Selecting analytical area→image→Duplicate
ImageJ operation : Duplicate stack
Example image
3. Making of background image
ImageJ operation : image→stacks→manupilation→substack maker
ImageJ operation
The number of the image used for the background is input
4. Making of difference picture
ImageJ operation : process→image calculator
ImageJ operation
Image1: File name of sequential image
Image2: File name of background image
Operation: subtract
5. Making of gray scale image
ImageJ operation : image→type→8bit
6. Adjusting contrasts
ImageJ operation : image→adjust→color balance
