Definition of deep multiphoton microscopy

Photonics Dictionary

deep multiphoton microscopy: Deep multiphoton microscopy is an advanced imaging technique used in biological and biomedical research to visualize structures and processes deep within tissues with high resolution. It combines the principles of multiphoton microscopy with specialized optics and imaging techniques to penetrate deeper into biological samples while minimizing photodamage and out-of-focus fluorescence.

In conventional fluorescence microscopy, excitation of fluorophores occurs through the absorption of a single photon of light, limiting penetration depth due to light scattering and absorption by biological tissues. In contrast, multiphoton microscopy utilizes the simultaneous absorption of two or more lower-energy photons to excite fluorescence. This nonlinear process enables deeper penetration into tissues compared to single-photon excitation techniques.

Deep multiphoton microscopy typically employs longer wavelength excitation light, such as NIR or IR light, which experiences less scattering and absorption in biological tissues compared to visible light. By using longer wavelengths and multiphoton excitation, deep multiphoton microscopy can achieve imaging depths of several hundred micrometers to several millimeters into tissues, depending on the specific sample and imaging parameters.

This imaging technique is particularly valuable for studying biological processes and structures in intact, thick tissues, such as brain imaging in neuroscience research, imaging of lymph nodes and organs in immunology and cancer research, and studying developmental processes in embryos and organs. Deep multiphoton microscopy provides researchers with the ability to visualize cellular and subcellular dynamics in their native tissue environment with high spatial resolution and minimal perturbation, advancing our understanding of complex biological systems.