Stimulated emission depletion (STED) microscopy enables the three-dimensional (3D) visualization of dynamic nanoscale structures in living cells, offering unique insights into their organization. However, 3D-STED imaging deep inside biological tissue is obstructed by optical aberrations and light scattering. We present a STED system that overcomes these challenges. Through the combination of two-photon excitation, adaptive optics, red-emitting organic dyes, and a long-working-distance water-immersion objective lens, our system achieves aberration-corrected 3D super-resolution imaging, which we demonstrate 164 µm deep in fixed mouse brain tissue and 76 µm deep in the brain of a living mouse.

中文翻译：

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活体小鼠的 3D 超分辨率深层组织成像

受激发射耗尽 (STED) 显微镜能够对活细胞中的动态纳米级结构进行三维 (3D) 可视化，从而提供对其组织的独特见解。然而，生物组织深处的 3D-STED 成像受到光学像差和光散射的阻碍。我们提出了一个克服这些挑战的 STED 系统。通过双光子激发、自适应光学、发红光的​​有机染料和长工作距离水浸物镜的组合，我们的系统实现了像差校正的 3D 超分辨率成像，我们展示了 164 µm 深度固定小鼠脑组织和 76 µm 深在活小鼠的大脑中。