We demonstrate the benefit of a novel laser strategy in multiphoton microscopy (MPM). The cheap, simple, and turn-key supercontinuum laser system with its spectral shaping module, constitutes an ideal approach for the one-shot microscopic imaging of many fluorophores without modification of the excitation parameters: central wavelength, spectral bandwidth, and average power. The polyvalence of the resulting multiplex-multiphoton microscopy (M-MPM) device is illustrated by images of many biomedical models from several origins (biological, medical, or vegetal), generated while keeping constant the spectral parameters of excitation. The resolution of the M-MPM device is quantified by a procedure of point-spread-function (PSF) assessment led by an original, robust, and reliable computational approach FIGARO. The estimated values for the PSF width for our M-MPM system are shown to be comparable to standard values found in optical microscopy. The simplification of the excitation system constitutes a significant instrumental progress in biomedical MPM, paving the way to the imaging of many fluorophores with a single shot of excitation without any modification of the lighting device.

中文翻译：

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用于生物医学成像的多重多光子显微镜和计算策略

我们展示了一种新型激光策略在多光子显微镜 (MPM) 中的好处。廉价、简单和交钥匙的超连续谱激光系统及其光谱整形模块构成了一种理想的方法，可以在不修改激发参数的情况下对许多荧光团进行一次显微成像：中心波长、光谱带宽和平均功率。由此产生的多重多光子显微镜 (M-MPM) 设备的多价性通过来自多个来源（生物、医学或植物）的许多生物医学模型的图像来说明，这些模型在保持激发光谱参数不变的情况下生成。M-MPM 设备的分辨率通过由原始、稳健且可靠的计算方法 FIGARO 领导的点扩展函数 (PSF) 评估程序进行量化。我们的 M-MPM 系统的 PSF 宽度估计值与光学显微镜中的标准值相当。激发系统的简化构成了生物医学 MPM 的重大仪器进步，为使用单次激发对许多荧光团进行成像铺平了道路，而无需对照明设备进行任何修改。