Multiphoton Microscopy for Brain Activity Imaging

Sato Masaaki, Azuma Kazuha, Iwatani Yu

doi:10.11410/kenbikyo.59.1_32

<p>Multiphoton excitation is a nonlinear optical process in which fluorescent molecules are excited by simultaneously absorbing multiple photons. Among microscopy techniques based on multiphoton excitation, two-photon microscopy (2PM) uses near-infrared pulsed lasers that penetrate well into highly light-scattering biological tissues and has exerted a profound impact on neuroscience, especially since the 2000s, as it allows for imaging of activity of complex neural circuits deep within the brain at single-cell resolution. On the other hand, typical 2PM scans the entire target volume with a focal point of about 1 μm³. This has limited its ability to observe the fast dynamics of neural circuit activity over a large brain area. To overcome this problem, recent 2PM takes advantage of various optical techniques to image a wider field of view at a faster sampling rate. Other technological innovations in multiphoton microscopy include three-photon microscopy to image the deeper brain more clearly, and miniaturized multiphoton microscopes that can be attached to the head of an animal to image brain circuit activity at a high resolution during freely-moving behavior.</p>

Multiphoton Microscopy for Brain Activity Imaging

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