NIH Image J

Prepared by:

K. Worth


Dow 2224


The following materials and equipment associated with this procedure present exposure or physical health hazards. Safety precautions are prudent and mandatory.



Prior to performing this procedure, the following safety equipment must be accessible and ready for use: (e.g. chemical fume hood, biological safety cabinet, laminar flow hood, chemical spill kits)


Protective Equipment:

Prior to performing this procedure, the following personal protective equipment must be obtained and ready for use: (e.g. acid resistant gloves, safety eyewear, lab coat, chemical splash apron)


Waste Disposal:

This procedure will result in the follow regulated waste which must be disposed of in compliance with environmental regulations.



In the event that a hazardous material spills during this procedure, be prepared to execute the emergency procedure below.

Not Applicable.


Note: Menu commands are in bold.

  1. Obtaining a clear image
    1. Using the optical microscope, select the area of your sample that you would like to analyze.
    2. Once you have found your selection, start up NIH Image.
    3. Pull down the special menu from the upper toolbar and select start capturing.
    4. Turn the optical assembly to the left (so the ocular lenses are pointing to your left). Your selection should appear in the window.
    5. Adjust the brightness or the focus on the microscope for the sharpest image possible.
    6. Pull down the special menu again and select stop capturing.
    7. Additional image manipulation can be accomplished by using the process menu. Use the sharpen or smooth command to further focus your image.
  2. Setting the scale NOTE: DEFAULT UNITS ARE IN PIXELS.
    1. Pull down the analyze menu and select set scale.
    2. From the units box, select appropriate units (default is pixels).
    3. Type in appropriate values for pixels and mm (or other unit).


Last Revision:

May 29, 2024