We have successfully developed a multiphoton-induced hyperspectral microscopy based on a 1064 nm femtosecond excitation source. Nonlinear excitation by the laser localized to the focal point allows efficient non-descanned detection (NDD) while achieving optically sectioned imaging. The use of 1064 nm laser excitation increases the imaging depth while minimizing sample damage. The system combines the advantages of NDD for 3D imagingrich spectral information through confocal hyperspectral imaging, leading to potential applications in the emerging material R&Dbiomedical research.

We combine the advantages of multiphoton microscopy with that of confocal microscopy to establish a hyperspectral multiphoton imaging system based on a 1064nm femtosecond excitation. The non-descanned detection allows efficient acquisition of optically sectioned images,the 1064nm laser provides increasing imaging depth while minimizing damage. The hyperspectral imaging reveals further details about the fluorescent origins. The system was designed from both the softwarethe hardware perspective to optimize the user＇s experience, allowing the user to focus on researchapplication.

Our system can provide 3D reconstructed imagingspectral information of intact tissues,can be used for in vivoin vitro studies to monitor celltissue dynamics. The ability of the system to monitor micron scale changes in live cellstissues at the depths of hundreds of micrometer makes it suitable for skin related applications such as transdermal drug delivery. The system is generally suitable any molecular cellular biology related research, such as stem cellcancer cell research. It can also be applied to investigate emerging materials such as 2D materials.

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