<p>A strongly focused laser beam with high power density employed in optical tweezers typically results in photothermal damage and instability to the trapped nanoparticles or bio-targets. Here, we report an effective laser cooling of trapped Yb³⁺-Er³⁺ co-doped rare-earth nanoparticle by employing a new cooling channel provided by Er³⁺ and utilizing an enhancement of the anti-Stokes emission through energy transfer between Yb³⁺ and Er³⁺. Especially in high temperature conditions, the co-doped system demonstrates a high laser cooling performance, that enable the temperature to decrease from 500 K to around room temperature. But for the initial temperature of around room temperature, the cooling temperature does not drop below freezing, which can avoid not only thermal but also cold damage to bio-targets in optical tweezers. This work provides experimental evidence of the Yb³⁺-Er³⁺ co-doping rare-earth nanoparticle for laser cooling, which can hold a potential to address the challenge of precise measurements in optical tweezers.</p>