How to use the ImageJ Particle Picker ------------------------------------- Open ImageJ from the desktop icon. From the FILE pull-down menu, select OPEN (open the next image in sequence). Select the image area by clicking and then dragging the crosshatch from the upper-left corner to the lower-right corner. From the IMAGE pull-down menu, select CROP. From the IMAGE pull-down menu, select ADJUST (brightness and contrast). Adjust the Contrast value to 222 (+ or -); adjust brightness as necessary. Click Apply and close window. From the IMAGE pull-down menu, select ADJUST-THRESHOLD. Adjust to highlight gold particles, but DO NOT CLOSE WINDOW or click on any other keys in the window. From the ANALYZE pull-down menu, in Set Measurements select "center of mass". From the PLUG IN pull-down menu, select PARTICLE PICKER. Click on circle icon; this displays ANALYZE PARTICLES. Set Size: 30 - 150 (for 5 - 6 nm gold particles), 100 - 150 (for 10 - 12 nm gold particles). Set Circularity: 0.00 - 1.00 Set Show: Nothing Unclick: Clear Results Select: Display Results Record Starts Click OK. Point list will be displayed. Minimize window. Reduce Threshold level to it's minimum (far left) value. Click on magnifying glass icon to zoom in (right click to zoom in, left click to zoom out). Click on pen icon to circle gold particles not circle automatically (right click) or erase a false reading (left click). You will need to go back and forth between the magnifying glass and pen as you scan the whole image. When complete, select the point list icon, which looks like a piece of paper. Save As: the image number excluding the date, with a code: experiment #-image #_gold particle size in nm (for example: 1881B-13_6). Close the image without saving changes. Close ImageJ, then reopen ImageJ for the next image. Open the saved file in notepad, then save it as a text file for Matlab analysis.