Research on field-enhanced planar-Airy light-sheet microscopy (invited)

Light-sheet microscopy is a biological imaging technology that has been studied a lot in recent years. Compared with traditional confocal laser scanning microscopy, light-sheet microscopy can achieve rapid volume imaging with low phototoxicity. The illumination beam of light-sheet microscope can choose Gaussian beam or other non-diffracting beams (such as Bessel beam, Airy beam, etc.). Airy light-sheet microscopy is the most researched technology at present, but there is a big problem with ordinary Airy light-sheet microscopy. Airy beam has the characteristic of self-bending, which causes Airy beam to exceed detection at both ends of the field of view. The depth of field of the objective lens cannot produce the best imaging effect. The Airy beam was rotated by 45° to form a planar-Airy light-sheet, so that the Airy light-sheet did not exceed the depth of field of the detection objective, so as to increase the imaging field of view of the light-sheet microscope. And using two-photon fluorescence excitation technology, the post-processing process of the image was eliminated, greatly improving the efficiency of imaging. In this study, Matlab was used for optical simulation, and the imaging field of view (~900 μm) of the planar-Airy light-sheet microscope was increased by 50% compared with the imaging field of view (~600 μm) of the ordinary Airy light-sheet microscope. The constructed planar-Airy light-sheet microscope was calibrated with fluorescent microspheres, and the lateral resolution of the imaging system was (1.93±0.17) μm and the axial resolution was (3.19±0.41) μm. In the real-time observation of the zebrafish intracerebral hemorrhage model, imaging results with a temporal resolution of x \times y \times z = 0.60 mm \times 0.60 mm \times 0.40 mm/60 s can be obtained, and the growth and development of local blood vessels can be monitored in real-time to explore the mechanism of cerebral hemorrhage disease.