β -Amyloid ( A β ) plaque, representing the progressive accumulation of the protein that mainly consists of A β , is one of the prominent pathological hallmarks of Alzheimer's disease (AD). Label-free imaging of A β plaques holds the potential to be a histological examination tool for diagnosing AD. We applied label-free multiphoton microscopy to identify extracellular A β plaque as well as intracellular A β accumulation for the first time from AD mouse models. We showed that a two-photon-excited fluorescence signal is a sensitive optical marker for revealing the spatial-temporal progression and the surrounding morphological changes of A β deposition, which demonstrated that both extracellular and intracellular A β accumulations play an important role in the progression of AD. Moreover, combined with a custom-developed image-processing program, we established a rapid method to visualize different degrees of A β deposition by color coding. These results provide an approach for investigating pathophysiology of AD that can complement traditional biomedical procedures.

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