We present two-photon fluorescence image scanning microscopy (ISM) with engineered excitation and detection point-spread-functions enabling 3D imaging in a single 2D scan. This demonstration combines excitation using a holographic multispot array of focused femtosecond pulses with a high-efficiency single-helix PSF phase mask detection. Camera detection along with a multiview reconstruction algorithm allows volumetric imaging of biological samples over a depth of field spanning more than 1500 nm with an axial resolution of better than 400 nm. The nonlinear two-photon process improves sectioning and the inherent longer wavelengths increase the penetration depth in scattering samples. Our method extends the performance of 3D ISM towards thicker biological samples.

中文翻译：

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双光子 PSF 工程图像扫描显微镜

我们展示了具有工程激发和检测点扩展功能的双光子荧光图像扫描显微镜 (ISM)，可在单个 2D 扫描中实现 3D 成像。该演示将使用聚焦飞秒脉冲的全息多点阵列激发与高效单螺旋 PSF 相位掩模检测相结合。相机检测和多视图重建算法允许在超过 1500 nm 的景深上对生物样本进行体积成像，轴向分辨率优于 400 nm。非线性双光子过程改善了切片，固有的较长波长增加了散射样品的穿透深度。我们的方法将 3D ISM 的性能扩展到更厚的生物样品。