The invention relates to a method and an apparatus for measuring the glass micro-bead refractive index. According to the present invention, when the refractive index of a high-refractive index glass micro-bead is measured, a reflecting mirror adjustment device and a camera five-dimensional adjustment device are adjusted so as to make the camera center, the light absorption center, and the hole center of the reflecting mirror having the hole coaxial, a single-mode He-Ne laser emits laser, the laser finally irradiates onto the glass micro-bead through a series of reflections and transmissions, a secondary rainbow image is obtained on a receiving screen, the camera collects the image, the receiving distance from the glass micro-bead to the receiving screen is measured, the radius of the outermost ring of the rainbow stripes is measured, the minimum deviation angle of the secondary rainbow of the glass micro-bead to be measured is calculated, and the refractive index is calculated through inversion of the minimum deviation angle; and when the refractive index of a low-refractive index glass micro-bead is measured, a spring locking device is opened, the instruments are adjusted so as to make the system in which the camera, the receiving screen and other devices exist rotate 90 DEG, the fixation is performed through a support frame, other steps are the same as the steps for measuring the refractive index of the high-refractive index glass micro-bead, and the minimum deviation angle of the primary rainbow of the glass micro-bead to be measured is calculated so as to calculate the refractive index through the inversion.