We demonstrated the feasibility of Raman spectroscopy for remote temperature monitoring within the aqueous humor of the rabbit eye in vivo. Using a confocal Raman spectroscopy system, Raman spectra from 2580 to 3800 cm −1 were recorded in HPLC-grade water and in the aqueous humor of the rabbit eye under in vivo and ex vivo conditions within a temperature range of 14-34°C. The ratio between the integrated Raman intensities of two temperature dependent OHvibrational regions (OH2/OH1) in the

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... 2/OH1) in the spectra of water showed high linear dependence on temperature both in pure water ͓0.0049 (Ϯ1.2%)Tϩ0.4522(Ϯ0.5%), R 2 ϭ0.99, nϭ50, pϽ0.05], as well as in the rabbit aqueous humor ͓0.0036(Ϯ2.8%)Tϩ0.4966(Ϯ0.6%), R 2 ϭ0.98, nϭ162, pϽ0.05] with a high degree of reproducibility and sensitivity (ϳ0.2-0.7°C). Raman spectroscopy can be used for high resolution and remote monitoring of temperature in the aqueous humor under in vivo conditions.