AbstractThis article discusses the applicability to a living animal of the temperature mapping method using the water proton chemical shift obtained with three‐dimensional magnetic resonance spectroscopic imaging (3D‐MRSI). There are several sources of error in obtaining the spectra with 3D‐MRSI: signal noise, limitation in the frequency resolution due to the finite signal length, intravoxel inhomogeneity in the static magnetic field, and variation in the magnetic field due to the eddy current magnetic field. A spectral estimation method called phase deduction complex Lorentzian fitting (PD‐CLF) was proposed. Numerical simulations demonstrated that this method reduces the error in the chemical shift to one third of that obtained with the simple frequency subtraction method that uses zero‐padded first Fourier transformation (FFT). The temperature images obtained using 3D‐MRSI with PD‐CLF clearly visualized the changes and distribution of temperature in an ane