Color-tunable upconversion luminescence has wide prospects for anti-counterfeiting and disease diagnosis/treatment. To date, achieving high-quality tunable red and blue emissions using a single excitation wavelength remains a formidable challenge, due to the large energy difference between the red and blue photons. In this Letter, based on Tm3+ upconversion luminescence, blue dominant and red dominant emissions are generated upon 980-nm excitations using a short and long pulse, respectively. The corresponding color tuning mechanisms are investigated based on the spectral observations. The proposed color tuning strategy is particularly useful for in vivo applications as the red and blue lights play important roles in biological imaging and drug release, respectively.