Abstract：
Volume electron microscopy (Volume EM) has emerged as a powerful tool for visualizing the 3D structure of cells and tissues with nanometer-level precision. Within the retina, various types of neurons establish synaptic connections in the inner and outer plexiform layers. While conventional EM techniques have yielded valuable insights into retinal subcellular organelles, their limitation lies in providing 2D image data, which can hinder accurate measurements. For instance, quantifying the size of three distinct synaptic vesicle pools, crucial for synaptic transmission, is challenging in 2D. Volume EM offers a solution by providing large-scale, high-resolution 3D data. It is worth noting that sample preparation is a critical step in Volume EM, significantly impacting image clarity and contrast. In this context, we outline a sample preparation protocol for the 3D reconstruction of photoreceptor axon terminals in the retina. This protocol includes three key steps: retina dissection and fixation, sample embedding processes, and selection of the area of interest.
摘要：
体积电子显微镜（VolumeEM）已成为一种强大的工具，用于以纳米级精度可视化细胞和组织的3D结构。在视网膜内，各种类型的神经元在内部和外部丛状层中建立突触连接。虽然传统的EM技术已经对视网膜亚细胞细胞器产生了有价值的见解，它们的局限性在于提供2D图像数据，这可能会阻碍准确的测量。例如,量化三个不同的突触小泡池的大小，对突触传递至关重要，在2D中具有挑战性。VolumeEM通过提供大规模、高分辨率3D数据。值得注意的是，样品制备是VolumeEM中的关键步骤，显着影响图像的清晰度和对比度。在这种情况下，我们概述了视网膜感光轴突末端三维重建的样品制备方案.该协议包括三个关键步骤：视网膜解剖和固定，样本嵌入过程，和选择感兴趣的领域。
