细胞生物系Joerg Bewersdorf研究组揭示了脑组织超微结构背景下特定分子的全光学可视化。该项研究成果发表在2025年11月26日出版的《自然—生物技术》上。

在这里，研究人员提出了组织泛扩展显微镜（pan-ExM-t），这是一种全光学成像方法，结合了~16 - 24倍线性扩展与蛋白质和脂质荧光泛染色（提供电子显微镜样的超微结构背景）和免疫标记（用于分子成像）。课题组通过在突触前和突触后密度的超微结构三维背景下成像突触和细胞特异性抗体，在分离的神经元培养物中成像神经纳米结构和细胞器，以及在无主题脑组织切片中成像，证明了这种方法的多功能性。

此外，课题组研究人员展示了从pan-ExM-t图像体积中追踪神经回路，这表明任何使用共聚焦显微镜的实验室现在都可以在纳米级细胞和电路环境中定位特定分子。

研究人员表示，了解大脑的分子解剖和神经连接需要成像技术，可以在大脑超微结构的背景下绘制特定蛋白质的三维纳米级分布。光学和电子显微镜可以可视化特定的标签或解剖超微结构，但将分子特异性与解剖背景相结合是具有挑战性的。

附：英文原文

Title: All-optical visualization of specific molecules in the ultrastructural context of brain tissue

Author: MSaad, Ons, Cairns, Allison, Gulcicek, Jonathan, Kasula, Ravi Kiran, Liao, Jacob, Kondratiuk, Ilona, Bewersdorf, Emma H., Kidd, Phylicia, Falahati, Hanieh, Gentile, Juliana E., Niescier, Robert F., Watters, Katherine, Sterner, Robert C., Lee, Seong-il, Guo, Xiaojia, Liu, Xinran, Desir, Gary, De Camilli, Pietro, Rothman, James E., Koleske, Anthony J., Biederer, Thomas, Kuan, Aaron T., Bewersdorf, Joerg

Issue&Volume: 2025-11-26

Abstract: Understanding the molecular anatomy and neural connectivity of the brain requires imaging technologies that can map the three-dimensional nanoscale distribution of specific proteins in the context of brain ultrastructure. Light and electron microscopy visualize either specific labels or anatomical ultrastructure but combining molecular specificity with anatomical context is challenging. Here we present pan-expansion microscopy of tissue (pan-ExM-t), an all-optical imaging method that combines ~16–24-fold linear expansion with fluorescent pan-stainings of proteins and lipids (providing electron microscopy-like ultrastructural context) and immunolabeling (for molecular imaging). We demonstrate the versatility of this approach by imaging synaptic and cell-specific antibodies in the ultrastructural three-dimensional context of presynaptic and postsynaptic densities, neuropil nanoarchitecture and cellular organelles in dissociated neuron cultures, and mouse brain tissue sections. Furthermore, we demonstrate tracing of neuronal circuitry from pan-ExM-t image volumes, suggesting that any laboratory with access to a confocal microscope can now localize specific molecules within nanoscale cellular and circuit contexts.

DOI: 10.1038/s41587-025-02905-4

Source: https://www.nature.com/articles/s41587-025-02905-4