Department of Biology, University of Maryland, College Park, MD 20742, USA.
Neuroscience and Cognitive Science Graduate Program, University of Maryland, College Park, MD 20742, USA.
STAR Protoc. 2021 Nov 24;2(4):100971. doi: 10.1016/j.xpro.2021.100971. eCollection 2021 Dec 17.
Here, we present a protocol for collecting large-volume, four-color, single-molecule localization imaging data from neural tissue. We have applied this technique to map the location and identities of chemical synapses across whole cells in mouse retinae. Our sample preparation approach improves 3D STORM image quality by reducing tissue scattering, photobleaching, and optical distortions associated with deep imaging. This approach can be extended for use on other tissue types enabling life scientists to perform volumetric super-resolution imaging in diverse biological models. For complete details on the use and execution of this protocol, please refer to Sigal et al. (2015).
在此，我们提供了一个从神经组织中收集大体积、四色、单分子定位成像数据的方案。我们已经将该技术应用于绘制小鼠视网膜全细胞中化学突触的位置和身份。我们的样品制备方法通过减少组织散射、光漂白和与深层成像相关的光畸变来提高 3D STORM 图像质量。这种方法可以扩展到其他组织类型的使用，使生命科学家能够在各种生物模型中进行体积超分辨率成像。有关此方案的使用和执行的完整详细信息，请参阅 Sigal 等人（2015 年）。
