This chapter will provide an overview of two-photon microscopy from elements of the theory underpinning fluorescence phenomena to functioning principles of a two-photon microscope including step-by-step practical advice on how to conduct an experiment using a two-photon microscope. In this context multi-photon excitation is also taken into consideration. By reading this chapter, you will have a synopsis of the basic principles of two-photon excitation, optical sectioning, and 3D microscopy. Furthermore, fundamentals of promising advanced methods for tissue imaging available for two-photon imaging as second harmonic generation (SHG) and fluorescence lifetime imaging microscopy (FLIM) are briefly described together with classical applications on deep tissue imaging and functional brain imaging.
Sancataldo, GiuseppeBarrera, Olga Vetri, Valeria
School of Engineering, Computing and Mathematics
Year of publication: 2022Date of RADAR deposit: 2023-03-13
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