Five-Modal Three-Dimensional Optical Microscopy via Single 1550 nm Fiber Laser

Abstract

Multimodal optical microscopy can be used to visualize multiple biological targets at the same location and at similar spatial resolutions, enabling more comprehensive studies of complex biological samples. However, current multimodal imaging systems typically use multiple or specially designed excitation lasers. Furthermore, previous studies have reported only three or four types of imaging modalities, limiting the number of imaging targets. This study develops an imaging system based on a single commercialized 1550 nm fiber laser and integrates five imaging modalities: second harmonic generation, third harmonic generation, two-photon fluorescence, three-photon fluorescence, and reflected confocal microscopy. Utilizing the characteristics of different imaging modalities, various biological components in biological tissues can be revealed simultaneously and cell movement in a complex biological environment can be traced noninvasively. The capacity of deep imaging has also been demonstrated in the mouse brain. The results show that the proposed multimodal microscopy system has great application potential in biomedical fields, such as tumor diagnosis and immunotherapy.