Abstract Multi-photon scanning light-sheet microscope, which combines the advantages of light-sheet illumination and multi-photon excitation, has high axial resolution, low photobleaching, and a large field of view. However, as the light sheet goes deeper into the biological tissue, biological optical aberrations caused by the uneven refractive index inside the organism become non-negligible factors that dramatically increase the thickness and decrease the light intensity of the light sheet. This paper theoretically deduces and simulates the influence of aberrations on the intensity and thickness of the two-, three-, and four-photon fluorescent light sheet. A clear general trend is obvious. With the increasing amplitude of aberration, the effect of aberration on the light sheet thickness and intensity increases in each multiphoton light sheet. In terms of thickness, the impact of high-order aberrations diminishes from the two-photon to the four-photon light sheet. However, in terms of intensity, the impact of high-order aberrations increases from the two-photon to the four-photon light sheet. The number of items to be corrected for each multiphoton light sheet is given and compared under random aberrations with different amplitudes. The results provide important guidance for the design and evaluation of adaptive optics in multi-photon light sheet microscopes.

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像差对多光子荧光灯片影响的模拟分析与比较

摘要 多光子扫描光片显微镜结合了光片照明和多光子激发的优点，具有轴向分辨率高、光漂白低、视场大等优点。然而，随着光片深入生物组织，生物体内折射率不均匀引起的生物光学像差成为不可忽视的因素，使光片的厚度急剧增加，光强度降低。本文从理论上推导并模拟了像差对二、三、四光子荧光灯片强度和厚度的影响。一个明显的大趋势是显而易见的。随着像差幅度的增加，像差对光片厚度和强度的影响在每个多光子光片中增加。在厚度方面，高阶像差的影响从双光子到四光子光片减弱。然而，在强度方面，高阶像差的影响从双光子到四光子光片增加。给出了每个多光子光片要校正的项目数，并在不同幅度的随机像差下进行比较。结果为多光子光片显微镜中自适应光学的设计和评估提供了重要指导。给出了每个多光子光片要校正的项目数，并在不同幅度的随机像差下进行比较。结果为多光子光片显微镜中自适应光学的设计和评估提供了重要指导。给出了每个多光子光片要校正的项目数，并在不同幅度的随机像差下进行比较。结果为多光子光片显微镜中自适应光学的设计和评估提供了重要指导。