The imaging method of SPECT (Single Photon Emission Computed Tomography) is used in nuclear medicine for diagnostics of various diseases or organs malfunctions. The distribution of medically injected, inhaled, or ingested radionuclides (radiotracers) in the patient body is imaged using gamma-ray sensitive camera with suitable imaging collimator. The 3D image is then calculated by combining many images taken from different observation angles. Most of SPECT systems use scintillator based cameras. These cameras do not provide good energy resolution and do not allow efficient suppression of unwanted signals such as those caused by Compton scattering. The main goal of this work is evaluation of Timepix3 detector properties for SPECT method for functional imaging of small animals during preclinical studies. Advantageous Timepix3 properties such as energy and spatial resolution are exploited for significant image quality improvement. Preliminary measurements were performed on specially prepared plastic phantom with cavities filled by radioisotopes and then repeated with in vivo mouse sample.