Fluorescein anisotropy, which is a widely used technique to study the folding state of proteins or affinity of ligands, is used in the present work to study the temperature sensing of fluid in a microchannel, by adding fluorophore in the fluid. Fluorescein was used as a temperature probe, while glycerol-aq. ammonia was used as a working fluid. Fluorescence anisotropy of fluorescein were measured by varying various parameters. Apart from this, a comparison of fluorescence anisotropy and fluorescence intensity is also performed.