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.
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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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