Significance Multiphoton microscopy is a powerful imaging tool for biomedical applications. A variety of techniques and respective benefits exist for multiphoton microscopy, but an enhanced resolution is especially desired. Additionally multiphoton microscopy requires ultrafast pulses for excitation, so optimization of the pulse duration at the sample is critical for strong signals. Aim We aim to perform enhanced resolution imaging that is robust to scattering using a structured illumination technique while also providing a rapid and easily repeatable means to optimize group delay dispersion (GDD) compensation through to the sample. Approach Spatial frequency modulation imaging (SPIFI) is used in two domains: the spatial domain (SD) and the wavelength domain (WD). The WD-SPIFI system is an in-line tool enabling GDD optimization that considers all material through to the sample. The SD-SPIFI system follows and enables enhanced resolution imaging. Results The WD-SPIFI dispersion optimization performance is confirmed with independent pulse characterization, enabling rapid optimization of pulses for imaging with the SD-SPIFI system. The SD-SPIFI system demonstrates enhanced resolution imaging without the use of photon counting enabled by signal to noise improvements due to the WD-SPIFI system. Conclusions Implementing SPIFI in-line in two domains enables full-path dispersion compensation optimization through to the sample for enhanced resolution multiphoton microscopy.

中文翻译：

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级联域多光子空间频率调制成像。

意义 多光子显微镜是生物医学应用的强大成像工具。多光子显微镜存在多种技术和各自的优点，但特别需要增强的分辨率。此外，多光子显微镜需要超快脉冲进行激发，因此优化样品的脉冲持续时间对于强信号至关重要。目标我们的目标是使用结构化照明技术执行对散射具有鲁棒性的增强分辨率成像，同时提供快速且易于重复的方法来优化样品的群延迟色散（GDD）补偿。方法 空间频率调制成像（SPIFI）用于两个域：空间域（SD）和波长域（WD）。WD-SPIFI 系统是一款在线工具，可实现 GDD 优化，考虑到样品的所有材料。SD-SPIFI 系统遵循并支持增强分辨率成像。结果 WD-SPIFI 色散优化性能通过独立的脉冲表征得到证实，从而能够快速优化 SD-SPIFI 系统成像的脉冲。SD-SPIFI 系统展示了增强的分辨率成像，无需使用光子计数，而 WD-SPIFI 系统通过信噪比改进实现了这一点。结论 在两个域中串联实施 SPIFI 可以实现对样品的全路径色散补偿优化，从而实现增强分辨率多光子显微镜。