A new strategy to prepare core/shell Janus photonic crystal (PC) microspheres with reversible optical spectrum property is demonstrated. The microfluidic technique was employed to generate the uniform core/shell PC microspheres containing nanogels aqueous suspension. Under centrifugal force, the nanogel particles homogeneously dispersed in the core of microspheres would aggregate in the half of the microspheres, leading to Janus PC microspheres with varied reflection spectra at the different side of the spheres. More interestingly, such Janus structure of PC microspheres and their reflection spectrum were significantly reversible when the centrifugation was employed and removed alternatively. In addition, due to the soft and thermal-responsive nature of the building blocks (e.g., nanogels), Janus structures and optical properties of the PC microspheres are highly influenced by the temperature, centrifugal speed, and time, providing the other parameters on the manipulation of properties of the PC microspheres. This strategy provides a new concept for the preparation of Janus PC microspheres with tunable structures and optical properties, which will find potential applications in the field of sensors, optical devices, barcodes, etc.