LED-based integrating sphere light sources (LED-ISLSs) in the size of typical microscope slides were developed to calibrate the radiance responsivity of optical imaging microscopes. Each LED-ISLS consists of a miniaturized integrating sphere with a diameter of 4 mm, an LED chip integrated on a printed circuit board, and a thin circular aperture with a diameter of 1 mm as the exit port. The non-uniformity of the radiant exitance of the LED-ISLSs was evaluated to be 0.8%. The normal radiance of the LED-ISLSs in the range of (5∼69) W m-2 sr-1 was measured with a standard uncertainty of 1.3% using two precision apertures and a standard silicon photodetector whose spectral responsivity is traceable to an absolute cryogenic radiometer. The LED-ISLSs were applied to calibrate the radiance responsivity of a home-built optical imaging microscope with a standard uncertainty of 2.6∼2.9%. The LED-ISLSs offer a practical way to calibrate the radiance responsivity of various optical imaging microscopes for results comparison and information exchange.