Second-harmonic generation (SHG) microscopy is a powerful label-free imaging tool widely used to visualize collagen and muscle in biological tissues. However, traditional laser-scanning SHG microscopy requiring voxel scanning is time-intensive. Wide-field SHG microscopy was designed to bypass this restriction, but its application to deep tissue imaging is limited due to vulnerability to scattering and sample-induced aberrations. We introduce synthetic aperture SHG (SA-SHG) microscopy to attenuate the effect of multiple scattering noises. Our SA-SHG method coherently integrates amplitude and phase maps of wide-field SHG fields taken for different illumination angles, thereby enhancing the signal-to-noise ratio. We also develop computational adaptive optics SHG (CAO-SHG) microscopy to computationally correct the sample-induced aberrations. Our algorithm optimizes SHG fields’ aperture synthesis to identify aberration maps, enabling the restoration of diffraction-limited imaging. We successfully apply this approach to real biological samples, demonstrating its potential for high-resolution imaging in complex biological environments.

中文翻译：

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具有合成孔径和计算自适应光学的二次谐波显微镜

二次谐波发生 (SHG) 显微镜是一种强大的无标记成像工具，广泛用于可视化生物组织中的胶原蛋白和肌肉。然而，需要体素扫描的传统激光扫描二次谐波显微镜非常耗时。宽视场二次谐波显微镜的设计就是为了绕过这一限制，但由于容易受到散射和样品引起的像差的影响，其在深层组织成像中的应用受到限制。我们引入合成孔径 SHG (SA-SHG) 显微镜来减弱多重散射噪声的影响。我们的 SA-SHG 方法相干地集成了不同照明角度下的宽视场 SHG 场的幅度和相位图，从而提高了信噪比。我们还开发了计算自适应光学二次谐波 (CAO-SHG) 显微镜，以计算校正样品引起的像差。我们的算法优化了 SHG 场的孔径合成来识别像差图，从而能够恢复衍射极限成像。我们成功地将这种方法应用于真实的生物样本，展示了其在复杂生物环境中高分辨率成像的潜力。