A measurement device of a central location of a single molecule using structured illumination and phase detection, the measurement device comprising: a light source configured to irradiate laser light; a light wavefront control unit configured to form the laser light irradiated by the light source into light having a predetermined periodic pattern to generate a structured illumination; a detection unit configured to detect optical signals emitted from the single molecule, which are generated from a target object to be generated by the structured illumination generated by the light wavefront control unit to acquire single molecule images; a light transmitting unit configured to transmit the laser light irradiated by the light source to the light wavefront control unit, transmit the structured illumination generated by the light wavefront control unit to the target object, and transmit the optical signals which are generated from the target object to be generated to the detection unit; and a controller configured to acquire single molecule images corresponding to the structured illumination whose phases are modulated by the light wavefront control unit while modulating the phase of the structured illumination generated by the light wavefront control unit and measure a central location of the target object based on a plurality of acquired single molecule images, wherein the light transmitting unit includes an optical fiber, a first polarization control plate for controlling a direction of linear polarization, a mirror, a polarization light separator, and a second polarization control plate for changing linear polarization and circular polarization to each other, which are sequentially arranged along a path of light such that the light transmitting unit changes the laser light irradiated by the light source into circular polarization state and transmits it to the light wavefront control unit. a light source configured to irradiate laser light; a light wavefront control unit configured to form the laser light irradiated by the light source into light having a predetermined periodic pattern to generate a structured illumination; a detection unit configured to detect optical signals emitted from the single molecule, which are generated from a target object to be generated by the structured illumination generated by the light wavefront control unit to acquire single molecule images; a light transmitting unit configured to transmit the laser light irradiated by the light source to the light wavefront control unit, transmit the structured illumination generated by the light wavefront control unit to the target object, and transmit the optical signals which are generated from the target object to be generated to the detection unit; and a controller configured to acquire single molecule images corresponding to the structured illumination whose phases are modulated by the light wavefront control unit while modulating the phase of the structured illumination generated by the light wavefront control unit and measure a central location of the target object based on a plurality of acquired single molecule images, wherein the light transmitting unit includes an optical fiber, a first polarization control plate for controlling a direction of linear polarization, a mirror, a polarization light separator, and a second polarization control plate for changing linear polarization and circular polarization to each other, which are sequentially arranged along a path of light such that the light transmitting unit changes the laser light irradiated by the light source into circular polarization state and transmits it to the light wavefront control unit.