Faculty of Medicine and Health, Charles Perkins Centre and School of Medical Sciences, The University of Sydney, Camperdown, NSW, Australia.
Biomedical Imaging Facility, Mark Wainwright Analytical Centre, University of New South Wales Sydney, Kensington, NSW, Australia.
J Neurosci Res. 2021 Jun;99(6):1515-1532. doi: 10.1002/jnr.24819. Epub 2021 Mar 7.
Ground state depletion followed by individual molecule return microscopy (GSDIM) has been used in the past to study the nanoscale distribution of protein co-localization in living cells. We now demonstrate the successful application of GSDIM to archival human brain tissue sections including from Alzheimer's disease cases as well as experimental tissue samples from mouse and zebrafish larvae. Presynaptic terminals and microglia and their cell processes were visualized at a resolution beyond diffraction-limited light microscopy, allowing clearer insights into their interactions in situ. The procedure described here offers time and cost savings compared to electron microscopy and opens the spectrum of molecular imaging using antibodies and super-resolution microscopy to the analysis of routine formalin-fixed paraffin sections of archival human brain. The investigation of microglia-synapse interactions in dementia will be of special interest in this context.
以往曾使用基态耗尽后单个分子返回显微镜（GSDIM）技术来研究活细胞中蛋白质共定位的纳米级分布。现在，我们成功地将 GSDIM 应用于存档的人脑组织切片，包括阿尔茨海默病病例以及来自小鼠和斑马鱼幼虫的实验组织样本。通过这种方法，可以在超越衍射极限的光镜分辨率下观察到突触前末端、小胶质细胞及其细胞过程，从而更清楚地了解它们在原位的相互作用。与电子显微镜相比，这里描述的方法节省了时间和成本，并将使用抗体和超分辨率显微镜的分子成像技术应用于分析存档的福尔马林固定石蜡包埋人脑的常规切片。在这种情况下，研究痴呆症中小胶质细胞-突触相互作用将特别有趣。
