Empathy is vital in interpersonal and societal interactions and serves as an emotional bridge fostering pro-social, including helping behavior. The process of offering help unfolds through distinct stages, starting with empathetic responses to another individual's negative emotional state, progressing to the decision to provide assistance, and culminating in the social rewards that follow such acts. Despite years of research in humans, the neural networks recruited in different stages of prosocial behavior still remain elusive. In this study, we established single photon emission computerized tomography (SPECT)-3D brain imaging protocol during ongoing helping behavior task (HBT) in mice. During this task, potential helper mice were tested over a period of six days to open a door and rescue their conspecifics trapped in a cold-water pool. On days 1, 3, and 5, CBF-SPECT imaging was conducted, whereas on days 2, 4, and 6, mice exclusively engaged in the HBT. We are unraveling the engagement of specific subcortical and cortical brain regions associated with empathy, motivational behavior, and reward processing at different stages of HBT. Additionally, we are establishing correlations between the activation of these brain regions and the extent of manifested helping behavior, while also exploring the active brain regions in individuals who do not exhibit helping behavior. Finally, the activation of brain areas involved in helping behavior in males and females will be discussed. In essence, our findings pave the way for future investigations into the causal roles of distinct brain regions at various stages of helping behavior.