In this paper, the application of a microsphere-based fiber-optic sensor with a 200 nm zinc oxide (ZnO) coating, deposited by the Atomic Layer Deposition (ALD) method, for temperature measurements between 100 and 300 °C, is presented. The main advantage of integrating a fiber-optic microsphere with a sensing device is the possibility of monitoring the integrity of the sensor head in real-time, which allows for higher accuracy during measurements. The study has demonstrated that ZnO ALD-coated microsphere-based sensors can be successfully used for temperature measurements. The sensitivity of the tested device was found to be 103.5 nW/°C when the sensor was coupled with a light source of 1300 nm central wavelength. The measured coefficient R2 of the sensor head was over 0.99, indicating a good fit of the theoretical linear model to the measured experimental data.