Abstract: The invention relates to a light calibration system consisting of a compact LED source with feedback control of intensity. The source is positioned in the focal plane of the microscope objective and produces flat-field illumination of up to 31 microwatts. The source can be easily used to determine the performance of microscope optics and camera response. It can also be used as a standard light source for calibration of experimental systems. Selectable light intensities are produced by controlling the LED input power via a feedback circuit consisting of a photodiode that detects output light intensity. Spectral coverage extends between 550 nm and 670 nm using green, yellow and red LEDS mounted side-by-side and which are individually selected. The LED chips are encapsulated in plastic diffusers which homogenize the light, and a flat field of illumination is obtained through a thin 1 mm diameter aperture positioned directly over each chip.