Abstract: With the constant development of multiphoton microscopy, our ability to observe complex and dynamic biological processes deeper within living tissue, is steadily improving. Researchers use multiphoton microscopy, because experiments can be conducted with little to no invasiveness or tissue damage over a long period of time with no photodamage (Mancuso et al., 2009). This allows for the introduction of tissue into the context of a three-dimensional (3D) environment in which visualization of cellular activation and interaction is viable. By circumventing a distorted reconstruction with limited z-stacks, multiphoton imaging provides enhanced spatiotemporal resolution.